49 results on '"Kenjiro Bandow"'
Search Results
2. CC chemokine ligand 20 (CCL20) positively regulates collagen type I production in 3D skin equivalent tissues
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Kenjiro Bandow, Avi Smith, Trishawna Watkins, Sasha Shenk, Behzad Gerami‐Naini, and Jonathan A. Garlick
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Dermatology ,Molecular Biology ,Biochemistry - Abstract
Chemokines are a group of small proteins that induce chemoattraction and inflammation and contribute to the differentiation and homeostasis of various cell types. Here we explored the role of chemokines, extracellular matrix production, and myofibroblast differentiation in self-assembled skin equivalents (SASE), a three-dimensional (3D) skin-equivalent tissue model. We found that the expression of three chemokines, C-C motif chemokine ligand (CCL) 20, C-X-C motif chemokine ligand (CXCL) 5, and CXCL8, increased with differentiation to myofibroblasts. Addition of recombinant CCL20 to human skin fibroblast induced collagen Type I alpha 2 gene expression, but did not affect the expression of alpha smooth muscle actin expression. Conversely, siRNA gene knockdown of CCL20 effectively inhibited the expression of collagen Type I gene and protein. Furthermore, when the CCL20 gene in fibroblasts was knocked down in SASE, collagen Type I synthesis and stromal thickness were decreased. Taken together, these results have indicated the utility of SASE in understanding how cytokines such as CCL20 positively regulate extracellular matrix proteins such as collagen Type I production during myofibroblast differentiation in 3D tissues that mimic human skin.
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- 2022
3. Dichloroacetyl Amides of 3,5-Bis(benzylidene)-4-piperidones Displaying Greater Toxicity to Neoplasms than to Non-Malignant Cells
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Mohammad Hossain, Praveen K. Roayapalley, Hiroshi Sakagami, Keitaro Satoh, Kenjiro Bandow, Umashankar Das, and Jonathan R. Dimmock
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General Engineering ,General Earth and Planetary Sciences ,unsaturated ketones ,cytotoxicity ,dichloroacetic acid ,tumor-selective toxicity ,structure-activity relationships ,General Environmental Science - Abstract
A series of 3,5-bis(benzylidene)-1-dichloroacetyl-4-piperidones 1a–l was evaluated against Ca9-22, HSC-2, HSC-3, and HSC-4 squamous cell carcinomas. Virtually all of the compounds displayed potent cytotoxicity, with 83% of the CC50 values being submicromolar and several CC50 values being in the double digit nanomolar range. The compounds were appreciably less toxic to human HGF, HPLF, and HPC non-malignant cells, which led to some noteworthy selectivity index (SI) figures. From these studies, 1d,g,k emerged as the lead molecules in terms of their potencies and SI values. A Quantitative Structure-Activity Relationship (QSAR) study revealed that cytotoxic potencies and potency–selectivity expression figures increased when the magnitude of the sigma values in the aryl rings was elevated. The modes of action of the representative cytotoxins in Ca9-22 cells were found to include G2/M arrest and stimulation of the cells to undergo mitosis and cause poly(ADP-ribose) polymerase (PARP) and procaspase 3 cleavage.
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- 2022
4. Glut1 expression is increased by p53 reduction to switch metabolism to glycolysis during osteoblast differentiation
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Tetsuya Matsuguchi, Kenjiro Bandow, Tomokazu Ohnishi, and Joji Kusuyama
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Cellular differentiation ,Gene Expression ,Oxidative phosphorylation ,Biochemistry ,Oxidative Phosphorylation ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Adenosine Triphosphate ,0302 clinical medicine ,Animals ,Glycolysis ,Molecular Biology ,030304 developmental biology ,Glucose Transporter Type 1 ,0303 health sciences ,Hexokinase ,Osteoblasts ,Kinase ,Cell Differentiation ,Cell Biology ,Cell biology ,Metabolic pathway ,chemistry ,Anaerobic glycolysis ,030220 oncology & carcinogenesis ,Tumor Suppressor Protein p53 ,Pyruvate kinase - Abstract
The glycolytic system is selected for ATP synthesis not only in tumor cells but also in differentiated cells. Differentiated osteoblasts also switch the dominant metabolic pathway to aerobic glycolysis. We found that primary osteoblasts increased expressions of glycolysis-related enzymes such as Glut1, hexokinase 1 and 2, lactate dehydrogenase A and pyruvate kinase M2 during their differentiation. Osteoblast differentiation decreased expression of tumor suppressor p53, which negatively regulates Glut1 expression, and enhanced phosphorylation of AKT, which is regulated by phosphoinositol-3 kinase (PI3K). An inhibitor of PI3K enhanced p53 expression and repressed Glut1 expression. Luciferase reporter assay showed that p53 negatively regulated transcriptional activity of solute carrier family 2 member 1 gene promoter region. Inhibition of glycolysis in osteoblasts reduced ATP contents more significantly than inhibition of oxidative phosphorylation by carbonyl cyanide m-chlorophenyl hydrazine. These results have indicated that osteoblasts increase Glut1 expression through the down-regulation of p53 to switch their metabolic pathway to glycolysis during differentiation.
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- 2020
5. In Vitro Assessment of Antitumor Potential and Combination Effect of Classical and Molecular-targeted Anticancer Drugs
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Yosuke Iijima, Takahiro Kaneko, Hiroshi Sakagami, Kenjiro Bandow, Shunsuke Hino, Norio Horie, and Motohiko Sano
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Drug ,Cisplatin ,Cancer Research ,education.field_of_study ,Cetuximab ,Chemistry ,Bortezomib ,media_common.quotation_subject ,Population ,General Medicine ,Pharmacology ,03 medical and health sciences ,0302 clinical medicine ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,Toxicity ,medicine ,Cytotoxicity ,education ,medicine.drug ,media_common - Abstract
Background/aim The aim of the study was to evaluate the antitumor potential and combination effects of chemotherapeutic drugs. Materials and methods The cytotoxicity of 20 drip-type classical and molecular-targeted anticancer drugs was examined against 4 human oral squamous cell carcinoma cell lines and 5 human oral normal mesenchymal and epithelial cells. Cell cycle progression was monitored by a cell sorter. Combination effect was evaluated by combination index. Results Most of the classical anticancer drugs showed much higher antitumor activity than molecular-targeted drugs, except bortezomib. Among 12 classical anticancer drugs, taxanes and gemsitabine showed the highest tumor-specificity (TS) and potency-selectivity expression (PSE) values, whereas platinum analogs showed the least TS value. Combination of two classical or a classical and a molecular-targeted drug showed mostly additive or antagonistic effect. 5-FU and cisplatin did not produce a subG1 population, but induced G2/M or G1/S arrest, regardless of the addition of cetuximab. Cetuximab, nibolumab and bortezomib showed potent keratinocyte toxicity. Conclusion The present TS monitoring system may provide useful information for building up the treatment regimens of anticancer drugs.
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- 2019
6. Quantitative Structure–Cytotoxicity Relationship of 2-Arylazolylchromones and 2-Triazolylchromones
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Koichi Takao, Hiroshi Sakagami, Junko Nagai, Haixia Shi, Kenjiro Bandow, Nao Yoshida, Mineko Tomomura, Natsuko Sezaki, Yoshiaki Sugita, Akito Tomomura, and Yoshihiro Uesawa
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Cancer Research ,Quantitative structure–activity relationship ,Cell Survival ,Stereochemistry ,Quantitative Structure-Activity Relationship ,Antineoplastic Agents ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Humans ,Viability assay ,Cytotoxicity ,Cell Proliferation ,Chemistry ,Cell Cycle ,General Medicine ,Cell cycle ,Oncology ,Chromones ,Cell culture ,030220 oncology & carcinogenesis ,Chromone ,Carcinoma, Squamous Cell ,Quantum Theory ,Mouth Neoplasms ,Drug Screening Assays, Antitumor ,Growth inhibition ,Lead compound - Abstract
BACKGROUND/AIM 4H-1-Benzopyran-4-one (chromone), present in various flavonoids as a backbone structure, has been used for the synthesis of anticancer drugs. The study aimed at investigating the cytotoxicity of eight 2-arylazolylchromones and twelve 2-triazolylchromones against four human oral squamous cell carcinoma (OSCC) cell lines and three human normal mesenchymal oral cells, and then performed a quantitative structure-activity relationship (QSAR) analysis. MATERIALS AND METHODS Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. The distribution of cells to various phases of cell cycle was determined by cell cycle analysis. A total of 3,218 physicochemical, structural and quantum chemical features were calculated for QSAR analysis from the most stabilized structure optimized using CORINA. RESULTS 2-[4-(4-fluorophenyl)-1H-imidazol-1-yl]-4H-1-benzopyran-4-one [6] had the highest tumor-specificity (TS), comparable with that of 5-flurouracil (5-FU) and doxorubicin, inducing cytostatic growth inhibition, accumulation of G2+M phase cells with no cells in the G1 phase. All eight 2-triazolylchromones showed much lower tumor-specificity, confirming our previous finding. Tumor-specificity was also correlated with 3D shape, topological shape, size, ionization potential, and the presence of more than two aromatic rings in the molecule and imidazole ring in the nitrogen-containing heterocyclic ring. CONCLUSION [6] can be a lead compound for designing anticancer drugs.
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- 2019
7. CXCL13 is a differentiation- and hypoxia-induced adipocytokine that exacerbates the inflammatory phenotype of adipocytes through PHLPP1 induction
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Tetsuya Matsuguchi, Kenjiro Bandow, Tomokazu Ohnishi, Kaori Shima, Joji Kusuyama, Muhammad Subhan Amir, and Ichiro Semba
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Male ,Adipose tissue ,Inflammation ,White adipose tissue ,Biochemistry ,Mice ,03 medical and health sciences ,Paracrine signalling ,0302 clinical medicine ,Adipokines ,3T3-L1 Cells ,Adipocytes ,Phosphoprotein Phosphatases ,medicine ,Animals ,Humans ,Hypoxia ,Autocrine signalling ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Gene knockdown ,Adipogenesis ,Adiponectin ,Interleukin-6 ,Chemistry ,Cell Biology ,Chemokine CXCL13 ,Cell biology ,Mice, Inbred C57BL ,PPAR gamma ,030220 oncology & carcinogenesis ,medicine.symptom - Abstract
Hypoxia in adipose tissue is regarded as a trigger that induces dysregulation of the secretory profile in adipocytes. Similarly, local dysregulation of adipocytokine secretion is an initial event in the deleterious effects of obesity on metabolism. We previously reported that CXCL13 is highly produced during adipogenesis, however little is known about the roles of CXCL13 in adipocytes. Here, we found that hypoxia, as modeled by 1% O2 or exposure to the hypoxia-mimetic reagent desferrioxamine (DFO) has strong inductive effects on the expression of CXCL13 and CXCR5, a CXCL13 receptor, in both undifferentiated and differentiated adipocytes and in organ-cultured white adipose tissue (WAT). CXCL13 was also highly expressed in WAT from high fat diet-fed mice. Hypoxic profile, typified by increased expression of interleukin-6 (IL-6) and plasminogen activator inhibitor-1 (PAI-1) and decreased expression of adiponectin, was significantly induced by CXCL13 treatment during adipogenic differentiation. Conversely, the treatment of adipocytes with a neutralizing-antibody against CXCL13 as well as CXCR5 knockdown by specific siRNA effectively inhibited DFO-induced inflammation. The phosphorylation of Akt2, a protective factor of adipose inflammation, was significantly inhibited by CXCL13 treatment during adipogenic differentiation. Mechanistically, CXCL13 induces the expression of PHLPP1, an Akt2 phosphatase, through focal adhesion kinase (FAK) signaling; and correspondingly we show that CXCL13 and DFO-induced IL-6 and PAI-1 expression was blocked by Phlpp1 knockdown. Furthermore, we revealed the functional binding sites of PPARγ2 and HIF1-α within the Cxcl13 promoter. Taken together, these results indicate that CXCL13 is an adipocytokine that facilitates hypoxia-induced inflammation in adipocytes through FAK-mediated induction of PHLPP1 in autocrine and/or paracrine manner.
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- 2019
8. Cytotoxic Tumour-Selective 1,5-Diaryl-3-Oxo-1,4-Pentadienes Mounted on a Piperidine Ring
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Praveen K. Roayapalley, Hiroshi Sakagami, Keitaro Satoh, Shigeru Amano, Kenjiro Bandow, Renato J. Aguilera, Karla G. Cano Hernandez, Austre Y. Schiaffino Bustamante, Stephen G. Dimmock, Rajendra K. Sharma, Umashankar Das, and Jonathan R. Dimmock
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unsaturated ketones ,General Engineering ,Medicine ,General Earth and Planetary Sciences ,cytotoxicity ,4-piperidones ,Western blot ,quantitative structure-activity relationships (QSAR) ,cell cycle inhibition ,Article ,General Environmental Science - Abstract
A series of 3,5-bis(benzylidene)-4-piperidones 2a–u were prepared as candidate cytotoxic agents. In general, the compounds are highly toxic to human gingival carcinoma (Ca9-22), human squamous carcinoma-2 (HSC-2) and human squamous carcinoma-4 (HSC-4) neoplasms, but less so towards non-malignant human gingival fibroblast (HGF), human periodontal ligament fibroblast (HPLF) and human pulp cells (HPC), thereby demonstrating tumour-selective toxicity. A further study revealed that most of the compounds in series 2 were more toxic to the human Colo-205 adenocarcinoma cell line (Colo-205), human HT29 colorectal adenocarcinoma cells (HT-29) and human CEM lymphoid cells (CEM) neoplasms than towards non-malignant human foreskin Hs27 fibroblast line (Hs27) cells. The potency of the cytotoxins towards the six malignant cell lines increased as the sigma and sigma star values of the aryl substituents rose. Attempts to condense various aryl aldehydes with 2,2,6,6-tetramethyl-4-piperidone led to the isolation of some 1,5-diaryl-1,4-pentadien-3-ones. The highest specificity for oral cancer cells was displayed by 2e and 2r. In the case of 2r, its selective toxicity exceeded that of doxorubicin and melphalan. The enones 2k, m, o have the highest SI values towards colon cancer and leukemic cells. Both 2e,r inhibited mitosis and increased the subG1 population (with a transient increase in G2/M phase cells). Slight activation of caspase-3, based on the cleavage of poly(ADP-ribose)polymerase (PARP) and procaspase 3, was detected.
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- 2021
9. Investigation of carbonic anhydrase inhibitory effects and cytotoxicities of pyrazole-based hybrids carrying hydrazone and zinc-binding benzenesulfonamide pharmacophores
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Cem Yamali, Hiroshi Sakagami, Keitaro Satoh, Kenjiro Bandow, Yoshihiro Uesawa, Silvia Bua, Andrea Angeli, Claudiu T. Supuran, and Halise Inci Gul
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Sulfonamides ,Molecular Structure ,Squamous Cell Carcinoma of Head and Neck ,Organic Chemistry ,Hydrazones ,Biochemistry ,Isoenzymes ,Structure-Activity Relationship ,Zinc ,Antigens, Neoplasm ,Head and Neck Neoplasms ,Drug Discovery ,Carcinoma, Squamous Cell ,Humans ,Pyrazoles ,Mouth Neoplasms ,Carbonic Anhydrase IX ,Carbonic Anhydrase Inhibitors ,Molecular Biology ,Carbonic Anhydrases - Abstract
Pyrazole-based carbohydrazone hybrids have been considered to be a remarkable class of compounds in pharmaceutical chemistry. Here, we reported bioactivities of 4-(3-(2-(arylidene)hydrazin-1-carbonyl)-5-phenyl-1H-pyrazol-1-yl)benzenesulfonamides (1-27) towards CA isoenzymes (hCA I, hCA II, hCA IX) and human oral squamous cell carcinoma cell line. Compounds 19 (Ki = 10.1 nM, hCA I/hCA IX = 749.6), 22 (Ki = 18.5 nM, hCA I/hCA IX = 429.2), 26 (Ki = 14.5 nM, hCA I/hCA IX = 596.9), 27 (Ki = 21.5 nM, hCA I/hCA IX = 413.1) were more potent and selective inhibitors of cancer-associated hCA IX isoenzyme. Compounds 22 and 26 were also found to be approximately three times more selective hCA IX inhibitors over off-target hCA II at low nanomolar. Compounds 19, 22, 23, 24, and 26 with IC
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- 2022
10. Purification and Biological Function of Caldecrin
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Kenjiro Bandow, Mineko Tomomura, and Akito Tomomura
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0301 basic medicine ,LPS ,Macrophage polarization ,Review ,macrophage ,calcium metabolism ,Bone resorption ,03 medical and health sciences ,0302 clinical medicine ,Osteoclast ,medicine ,TREM2 ,TLR4 ,Receptor ,General Environmental Science ,biology ,Chemistry ,Activator (genetics) ,General Engineering ,RANKL ,protease ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,osteoclast ,biology.protein ,Medicine ,General Earth and Planetary Sciences ,bone metabolism ,Signal transduction - Abstract
Blood calcium homeostasis is critical for biological function. Caldecrin, or chymotrypsin-like elastase, was originally identified in the pancreas as a serum calcium-decreasing factor. The serum calcium-decreasing activity of caldecrin requires the trypsin-mediated activation of the protein. Protease activity-deficient mature caldecrin can also reduce serum calcium concentration, indicating that structural processing is necessary for serum calcium-decreasing activity. Caldecrin suppresses the differentiation of bone-resorbing osteoclasts from bone marrow macrophages (BMMs) by inhibiting receptor activator of NF-κB ligand (RANKL)-induced nuclear factor of activated T-cell cytoplasmic 1 expression via the Syk–PLCγ–Ca2+ oscillation-calcineurin signaling pathway. It also suppresses mature osteoclastic bone resorption by RANKL-stimulated TRAF6–c-Src–Syk–calcium entry and actin ring formation. Caldecrin inhibits lipopolysaccharide (LPS)-induced osteoclast formation in RANKL-primed BMMs by inducing the NF-κB negative regulator A20. In addition, caldecrin suppresses LPS-mediated M1 macrophage polarization through the immunoreceptor triggering receptor expressed on myeloid cells (TREM) 2, suggesting that caldecrin may function as an anti-osteoclastogenic and anti-inflammatory factor via TREM2. The ectopic intramuscular expression of caldecrin cDNA prevents bone resorption in ovariectomized mice, and the administration of caldecrin protein also prevents skeletal muscle destruction in dystrophic mice. In vivo and in vitro studies have indicated that caldecrin is a unique multifunctional protease and a possible therapeutic target for skeletal and inflammatory diseases.
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- 2021
11. Quantitative Structure–Cytotoxicity Relationship of Azulene Amide Derivatives
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Noriyuki Okudaira, Akito Tomomura, Kana Imanari, Hiroshi Sakagami, Yoshihiro Uesawa, Hidetsugu Wakabayashi, Kenjiro Bandow, Junko Nagai, Mineko Tomomura, and Masashi Hashimoto
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Cancer Research ,Azulene amides ,Cell Survival ,Stereochemistry ,Population ,Cell ,Quantitative Structure-Activity Relationship ,Antineoplastic Agents ,Apoptosis ,Azulenes ,Cell Line ,chemistry.chemical_compound ,medicine ,Humans ,Cytotoxic T cell ,Cytotoxicity ,education ,hydrophobicity ,education.field_of_study ,QSAR ,Chemistry ,chemical descriptor ,General Medicine ,Azulene ,Amides ,oral squamous cell carcinoma ,medicine.anatomical_structure ,Oncology ,Cell culture ,molecular shape ,Carcinoma, Squamous Cell ,Mouth Neoplasms ,Lead compound ,apoptosis tumor specificity - Abstract
Background/Aim: Very few studies of anticancer activity of azulene amides led us to investigate the cytotoxicity of 21 N-alkylazulene-1-carboxamides introduced either with 3-methyl [1-7], 7-isopropyl-3-methyl [8-14] or 2-methoxy group [15-21]. Materials and Methods: Tumor-specificity (TS) was calculated by the ratio of mean 50% cytotoxic concentration (CC50) against three normal human oral mesenchymal cells to that against four human oral squamous cell carcinoma (OSCC) cell lines. Potency-selectivity expression (PSE) was calculated by dividing TS value by CC50 value against OSCC cell lines. Apoptosis-inducing activity was evaluated by caspase-3 activation and appearance of subG1 cell population. Results: [8-14] showed higher TS and PSE values, than [1-7] and [15-21]. The most active compound [8-14] induced apoptosis in C9-22 OSCC cells at 4-times higher CC50. Quantitative structure-activity relationship analysis of [1-14] demonstrated that their tumor-specificity was correlated with chemical descriptors that explain the molecular shape and hydrophobicity. Conclusion: 7-Isopropyl-3-methyl-N-propylazulene-1-carboxamide [8] can be a potential candidate of lead compound for manufacturing new anticancer drug.
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- 2019
12. JNK inactivation suppresses osteogenic differentiation, but robustly induces osteopontin expression in osteoblasts through the induction of inhibitor of DNA binding 4 (Id4)
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Brent G. Albertson, Kenjiro Bandow, Muhammad Subhan Amir, Kaori Shima, Toshiaki Nakamura, Joji Kusuyama, Kazuyuki Noguchi, Tetsuya Matsuguchi, Ichiro Semba, and Tomokazu Ohnishi
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musculoskeletal diseases ,0301 basic medicine ,MAP Kinase Signaling System ,Osteocalcin ,Biochemistry ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Immune system ,Osteogenesis ,Genetics ,Animals ,Mitogen-Activated Protein Kinase 9 ,Protein Isoforms ,Bone formation ,Osteopontin ,RNA, Small Interfering ,Molecular Biology ,Cells, Cultured ,Mice, Knockout ,Osteoblasts ,biology ,Chemistry ,JNK Mitogen-Activated Protein Kinases ,Gene Expression Regulation, Developmental ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,biology.protein ,Dual-Specificity Phosphatases ,Mitogen-Activated Protein Kinase Phosphatases ,Inhibitor of Differentiation Proteins ,RNA Interference ,030217 neurology & neurosurgery ,DNA ,Biotechnology - Abstract
Osteoblasts are versatile cells involved in multiple whole-body processes, including bone formation and immune response. Secretory amounts and patterns of osteoblast-derived proteins such as osteopontin (OPN) and osteocalcin (OCN) modulate osteoblast function. However, the regulatory mechanism of OPN and OCN expression remains unknown. Here, we demonstrate that p54/p46 c-jun N-terminal kinase (JNK) inhibition suppresses matrix mineralization and OCN expression but increases OPN expression in MC3T3-E1 cells and primary osteoblasts treated with differentiation inducers, including ascorbic acid, bone morphogenic protein-2, or fibroblast growth factor 2. Preinhibition of JNK before the onset of differentiation increased the number of osteoblasts that highly express OPN but not OCN (OPN-OBs), indicating that JNK affects OPN secretory phenotype at the early stage of osteogenic differentiation. Additionally, we identified JNK2 isoform as being critically involved in OPN-OB differentiation. Microarray analysis revealed that OPN-OBs express characteristic transcription factors, cell surface markers, and cytokines, including glycoprotein hormone α2 and endothelial cell-specific molecule 1. Moreover, we found that inhibitor of DNA binding 4 is an important regulator of OPN-OB differentiation and that dual-specificity phosphatase 16, a JNK-specific phosphatase, functions as an endogenous regulator of OPN-OB induction. OPN-OB phenotype was also observed following LPS from
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- 2019
13. Antitumor Effects and Tumor-specificity of Guaiazulene-3-Carboxylate Derivatives Against Oral Squamous Cell Carcinoma
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Hiroshi Sakagami, Masakazu Fujise, Yoshihiro Uesawa, Noriyuki Okudaira, Kenjiro Bandow, Michito Teratani, Shouta Nakamura, Junko Nagai, Masashi Hashimoto, Yosuke Iijima, and Hidetsugu Wakabayashi
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Cancer Research ,Cell Survival ,Quantitative Structure-Activity Relationship ,Antineoplastic Agents ,Apoptosis ,Azulenes ,Cell Line ,03 medical and health sciences ,chemistry.chemical_compound ,Sesquiterpenes, Guaiane ,0302 clinical medicine ,tumor-specificity ,Cell Line, Tumor ,Guaiazulene ,Cytotoxic T cell ,Humans ,Carboxylate ,Molecular Structure ,Chemistry ,QSAR ,Mesenchymal stem cell ,Cell Cycle ,General Medicine ,guaiazulene-3-carboxylate ,In vitro ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,molecular shape ,Alkoxy group ,Cancer research ,Carcinoma, Squamous Cell ,Amine gas treating ,ionization potential ,Mouth Neoplasms ,OSCC - Abstract
Aim: The aim of this study was to investigate the antitumor potential of guaiazulene-3-carboxylate derivatives against oral malignant cells. Materials and Methods: Twelve guaiazulene-3-carboxylate derivatives were synthesized by introduction of either with alkyl group [1-5], alkoxy group [6, 7], hydroxyl group [8, 9] or primary amine [10-12] at the end of sidechains. Tumor-specificity (TS) was calculated by the ratio of mean 50% cytotoxic concentration (CC50) against 3 human oral mesenchymal cell lines to that against 4 human oral squamous cell carcinoma (OSCC) cell lines. Potency-selectivity expression (PSE) was calculated by dividing TS value by CC50value against OSCC cell lines. Cell cycle analysis was performed by cell sorter. Results: [6, 7] showed the highest TS and PSE values, and induced the accumulation of both subG1 and G2/M cell populations in HSC-2 OSCC cells. Quantitative structure-activity relationship analysis demonstrated that their tumor-specificity was correlated with chemical descriptors that explain the 3D shape, electric state and ionization potential. Conclusion: Alkoxyl guaiazulene-3-carboxylates [6, 7] can be potential candidates of lead compound for developing novel anticancer drugs., Permission Granted by Publisher
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- 2020
14. Protection of Bortezomib-induced Neurotoxicity by Antioxidants
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Takahiro Kaneko, Motohiko Sano, Shunsuke Hino, Norio Horie, Kenjiro Bandow, Shigeru Amano, Hiroshi Sakagami, and Yosuke Iijima
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Cancer Research ,medicine.medical_treatment ,Cellular differentiation ,Antineoplastic Agents ,Apoptosis ,Pharmacology ,PC12 Cells ,Antioxidants ,Bortezomib ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Nerve Growth Factor ,medicine ,Cytotoxic T cell ,Animals ,Humans ,Cytotoxicity ,Cisplatin ,Neurons ,Chemotherapy ,business.industry ,Cell Cycle ,Peripheral Nervous System Diseases ,Cell Differentiation ,General Medicine ,Cell cycle ,Rats ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,Carcinoma, Squamous Cell ,Mouth Neoplasms ,Neurotoxicity Syndromes ,business ,medicine.drug - Abstract
Background/aim Although chemotherapy agents, such as oxaliplatin, cisplatin, paclitaxel and bortezomib frequently cause severe peripheral neuropathy, very few studies have reported the effective strategy to prevent this side effect. In this study, we first investigated whether these drugs show higher neuropathy compared to a set of 15 other anticancer drugs, and then whether antioxidants, such as sodium ascorbate, N-acetyl-L-cysteine, and vitamin B12 have any protective effect against them. Materials and methods Rat PC12 cells were induced to differentiate into neuronal cells by repeated overlay of serum-free medium supplemented with nerve growth factor. The cytotoxic levels of anticancer drugs against four human oral squamous cell carcinoma cell lines, three normal oral cells, and undifferentiated and differentiated PC12 cells were determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Cells were sorted for apoptotic cells (distributed into subG1 phase) and cells at different stages of cell cycle (G1, S and G2/M). Results All 19 anticancer drugs showed higher cytotoxicity against PC12 compared to oral normal cells. Among them, bortezomib showed the highest cytotoxicity against both undifferentiated and differentiated PC12 cell and, committed them to undergo apoptosis. Sodium ascorbate and N-acetyl-L-cysteine, but not vitamin B12, completely reversed the cytotoxicity of bortezomib. Conclusion Bortezomib-induced neuropathy might be ameliorated by antioxidants.
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- 2020
15. Biological Properties of the Aggregated Form of Chitosan Magnetic Nanoparticle
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Yosuke Iijima, Shinsuke Ifuku, Hiroshi Sakagami, Yukari Koga-Ogawa, Tadamasa Nobesawa, René García-Contreras, Satoshi Yokose, Angel David Paulino-Gonzalez, Yumiko Kanda, Giichirou Nakaya, Yasushi Hibino, Osamu Amano, Sachie Nakatani, Daisuke Ueda, Hiroshi Nakajima, Kenjiro Bandow, Akiyoshi Shiroto, Kenji Kobata, Yuko Nagasawa, and Masaji Yamamoto
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Cancer Research ,Biocompatibility ,Population ,General Biochemistry, Genetics and Molecular Biology ,Chitosan ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Drug Delivery Systems ,Humans ,MTT assay ,Viability assay ,Particle Size ,Cytotoxicity ,education ,Magnetite Nanoparticles ,Pharmacology ,education.field_of_study ,Drug Carriers ,Chemistry ,030220 oncology & carcinogenesis ,Drug delivery ,Biophysics ,Nanoparticles ,Iron oxide nanoparticles ,Research Article - Abstract
Background/Aim: Chitosan-coated iron oxide nanoparticles (Chi-NP) have gained attention because of their biocompatibility, biodegradability, low toxicity and targetability under magnetic field. In this study, we investigated various biological properties of Chi-NP. Materials and Methods: Chi-NP was prepared by mixing magnetic NP with chitosan FL-80. Particle size was determined by scanning and transmission electron microscopes, cell viability by MTT assay, cell cycle distribution by cell sorter, synergism with anticancer drugs by combination index, PGE(2) production in human gingival fibroblast was assayed by ELISA. Results: The synthetic process of Chi-NP from FL-80 and magnetic NP increased the affinity to cells, up to the level attained by nanofibers. Upon contact with the culture medium, Chi-NP instantly formed aggregates and interfered with intracellular uptake. Aggregated Chi-NP did not show cytotoxicity, synergism with anticancer drugs, induce apoptosis (accumulation of subG1 cell population), protect the cells from X-ray-induced damage, nor affected both basal and IL-1β-induced PGE(2) production. Conclusion: Chi-NP is biologically inert and shows high affinity to cells, further confirming its superiority as a scaffold for drug delivery.
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- 2020
16. Caldecrin inhibits lipopolysaccharide-induced pro-inflammatory cytokines and M1 macrophage polarization through the immunoreceptor triggering receptor expressed in myeloid cells-2
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Mineko Tomomura, Kenjiro Bandow, Akito Tomomura, and Hiroya Hasegawa
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0301 basic medicine ,Lipopolysaccharides ,Lipopolysaccharide ,Biophysics ,Macrophage polarization ,Biochemistry ,Proinflammatory cytokine ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Osteoclast ,medicine ,Animals ,Receptors, Immunologic ,Receptor ,Molecular Biology ,Membrane Glycoproteins ,biology ,Base Sequence ,Activator (genetics) ,Macrophages ,Serine Endopeptidases ,Cell Polarity ,Cell Biology ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,RAW 264.7 Cells ,chemistry ,Gene Expression Regulation ,RANKL ,030220 oncology & carcinogenesis ,biology.protein ,Cytokines ,Signal transduction ,Inflammation Mediators ,Signal Transduction - Abstract
Caldecrin was previously isolated as a serum calcium-decreasing factor from the pancreas and is known to suppress receptor activator of nuclear factor-κB ligand (RANKL)-induced calcium oscillation pathways in osteoclasts. Here, we explored the effects of caldecrin on lipopolysaccharide (LPS)–Toll-like receptor-4 (TLR-4) signaling pathways in macrophages. Caldecrin inhibited the LPS-induced gene expression of pro-inflammatory cytokines and M1 macrophage polarization in mouse bone marrow macrophages and the RAW264.7 mouse macrophage cell line. Next, we focused on triggering receptor expressed in myeloid cells-2 (TREM-2) as a co-receptor common to RANKL receptor and TLR-4, and established Trem2-KO RAW264.7 cells, in which Trem2 gene was deleted using the CRISPR/Cas9 system. Caldecrin-mediated alterations in pro-inflammatory cytokine expression and M1 macrophage polarization were not observed in Trem2-KO RAW264.7 cells. These results suggest that caldecrin is not only an inhibitor of osteoclast activation but also a negative regulator of LPS-induced inflammatory responses, functioning via TREM-2.
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- 2019
17. Quantitative Structure–Cytotoxicity Relationship of 3-(N-Cyclicamino)chromone Derivatives
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Koichi Takao, Kenjiro Bandow, Yoshihiro Uesawa, Noriyuki Okudaira, Tsukasa Sakatsume, Hiroshi Sakagami, Haixia Shi, Yoshiaki Sugita, Junko Nagai, Mineko Tomomura, and Akito Tomomura
- Subjects
Cancer Research ,Quantitative structure–activity relationship ,010405 organic chemistry ,Biological activity ,General Medicine ,01 natural sciences ,Molecular biology ,Chemical synthesis ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,chemistry.chemical_compound ,Oncology ,chemistry ,Cell culture ,Lipophilicity ,Chromone ,Growth inhibition ,Cytotoxicity - Abstract
BACKGROUND/AIM 4H-1-Benzopyran-4-ones (chromones) provide a backbone structure for the chemical synthesis of potent anticancer drugs. In contrast to 2-(N-cyclicamino)chromones, the biological activity of 3-(N-cyclicamino)chromones has not been reported. In this study, cytotoxicity of 15 3-(N-cyclicamino)chromone derivatives was investigated and subjected to quantitative structure-activity relationship (QSAR) analysis. MATERIALS AND METHODS Cytotoxicity against four human oral squamous cell carcinoma cell lines and three oral normal mesenchymal cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Tumor-specificity (TS) was evaluated as the ratio of mean 50% cytotoxic concentration (CC50) against normal oral cells to that against human oral squamous cell carcinoma cell lines. Potency-selectivity expression (PSE) value was calculated by dividing the TS value by the CC50 against tumor cells. Apoptosis induction was evaluated by morphological observation, western blot analysis and cell-cycle analysis. For QSAR analysis, a total of 3,096 physicochemical, structural and quantum chemical features were calculated from the most stabilized structure optimized using CORINA. RESULTS 3-(4-phenyl-1-piperazinyl)-4H-1-benzopyran-4-one (3a) had the highest tumor specificity, comparable with that of melphalan, without induction of apoptosis. Compound 3a caused cytostatic growth inhibition and had much lower cytotoxicity against human oral keratinocytes compared to doxorubicin. TS of the 15 3-(N-cyclicamino)chromones was correlated with 3D structure and lipophilicity. CONCLUSION Chemical modification of 3a may be a potential choice for designing a new type of anticancer drug.
- Published
- 2018
18. Quantitative Structure–Cytotoxicity Relationship of Pyrano[4,3-b]chromones
- Author
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Yoshihiro Uesawa, Kenjiro Bandow, Noriyuki Okudaira, Haixia Shi, Yuka Kubota, Hiroshi Sakagami, Junko Nagai, Mineko Tomomura, Akito Tomomura, Koichi Takao, and Yoshiaki Sugita
- Subjects
Cancer Research ,Quantitative structure–activity relationship ,Chemistry ,Biological activity ,General Medicine ,01 natural sciences ,Molecular biology ,Chemical synthesis ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Oncology ,Apoptosis ,Cell culture ,030220 oncology & carcinogenesis ,Chromone ,Growth inhibition ,Cytotoxicity - Abstract
BACKGROUND/AIM 4H-1-Benzopyran-4-one (chromone) provides a backbone structure for the chemical synthesis of potent anticancer drugs. Since studies of the biological activity of pyrano[4,3-b]chromones are limited, we investigated a total of 20 pyrano[4,3-b]chromones (10 sets of diastereomers) for their cytotoxicity against four human oral squamous cell carcinoma (OSCC) cell lines and human normal oral cells, and then carried out a quantitative structure-activity relationship (QSAR) analysis. MATERIALS AND METHODS Cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Tumor-specificity (TS) was evaluated by the ratio of mean 50% cytotoxic concentration (CC50) against normal oral cells to that against human OSCC cell lines. Potency-selectivity expression (PSE) value was calculated by dividing the TS value by the CC50 against tumor cells. Apoptosis induction was evaluated by morphological observation, western blot analysis and cell-cycle analysis. For QSAR analysis, a total of 3,072 physicochemical, structural and quantum chemical features were calculated from the most stabilized structure optimized using CORINA. RESULTS 8-Chloro-4,4a-dihydro-3-methoxy-3-methyl-3H,10H-pyrano[4,3-b][1]benzopyran-10-one (16) and 3-ethoxy-4,4a-dihydro-8-methoxy-3H,10H-pyrano[4,3-b][1]benzopyran-10-one (17) had the highest TS, higher than that of 5-flurouracil and melphalan, without induction of apoptosis. Compound 16 induced cytostatic growth inhibition and much lower cytotoxicity against human normal oral keratinocytes compared to doxorubicin. TS of 20 pyrano[4,3-b]chromones was correlated with 3D structure, polarity, ionic potential and electric state. CONCLUSION Chemical modification of 16 may be a potential choice for designing a new type of anticancer drug.
- Published
- 2018
19. Spleen tyrosine kinase influences the early stages of multilineage differentiation of bone marrow stromal cell lines by regulating phospholipase C gamma activities
- Author
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Joji Kusuyama, Tomokazu Ohnishi, Tetsuya Matsuguchi, Seong ChangHwan, Nahoko Eiraku, Ai Kamisono, Kenjiro Bandow, and Muhammad Subhan Amir
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Time Factors ,Stromal cell ,Physiology ,medicine.medical_treatment ,Clinical Biochemistry ,Syk ,Transfection ,Cell Line ,03 medical and health sciences ,Osteogenesis ,Internal medicine ,Enhancer binding ,Adipocytes ,medicine ,Animals ,Humans ,Syk Kinase ,Cell Lineage ,RNA, Messenger ,Phosphorylation ,Adaptor Proteins, Signal Transducing ,GRB2 Adaptor Protein ,Mice, Inbred C3H ,Adipogenesis ,Osteoblasts ,030102 biochemistry & molecular biology ,Phospholipase C gamma ,Chemistry ,Growth factor ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,hemic and immune systems ,Cell Biology ,Cell biology ,RUNX2 ,Phenotype ,030104 developmental biology ,Endocrinology ,Gene Expression Regulation ,RNA Interference ,Signal transduction ,Biomarkers ,Signal Transduction - Abstract
Bone marrow stromal cells (BMSCs) are multipotent cells that can differentiate into adipocytes and osteoblasts. Inadequate BMSC differentiation is occasionally implicated in chronic bone metabolic disorders. However, specific signaling pathways directing BMSC differentiation have not been elucidated. Here, we explored the roles of spleen tyrosine kinase (Syk) in BMSC differentiation into adipocytes and osteoblasts. We found that Syk phosphorylation was increased in the early stage, whereas its protein expression was gradually decreased during the adipogenic and osteogenic differentiation of two mouse mesenchymal stromal cell lines, ST2 and 10T(1/2), and a human BMSC line, UE6E-7-16. Syk inactivation with either a pharmacological inhibitor or Syk-specific siRNA suppressed adipogenic differentiation, characterized by decreased lipid droplet appearance and the gene expression of fatty acid protein 4 (Fabp4), peroxisome proliferator-activated receptor γ2 (Pparg2), CCAAT/enhancer binding proteins α (C/EBPα), and C/EBPβ. In contrast, Syk inhibition promoted osteogenic differentiation, represented by increase in matrix mineralization and alkaline phosphatase (ALP) activity, as well as the expression levels of osteocalcin, runt-related transcription factor 2 (Runx2), and distal-less homeobox 5 (Dlx5) mRNAs. We also found that Syk-induced signals are mediated by phospholipase C γ1 (PLCγ1) in osteogenesis and PLCγ2 in adipogenesis. Notably, Syk-activated PLCγ2 signaling was partly modulated through B-cell linker protein (BLNK) in adipogenic differentiation. On the other hand, growth factor receptor-binding protein 2 (Grb2) was involved in Syk-PLCγ1 axis in osteogenic differentiation. Taken together, these results indicate that Syk-PLCγ signaling has a dual role in regulating the initial stage of adipogenic and osteogenic differentiation of BMSCs.
- Published
- 2017
20. Osteopontin inhibits osteoblast responsiveness through the down-regulation of focal adhesion kinase mediated by the induction of low–molecular weight protein tyrosine phosphatase
- Author
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Tomokazu Ohnishi, Joji Kusuyama, Kenjiro Bandow, Tetsuya Matsuguchi, Kaori Shima, Mitsuhiro Hisadome, and Ichiro Semba
- Subjects
0301 basic medicine ,Cell Physiology ,Platelet-derived growth factor ,PTK2 ,Down-Regulation ,Protein tyrosine phosphatase ,Biology ,Calcitriol receptor ,Focal adhesion ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,stomatognathic system ,Cell Movement ,Cell Adhesion ,medicine ,Animals ,Osteopontin ,Phosphorylation ,Molecular Biology ,Platelet-Derived Growth Factor ,Osteoblasts ,Cell Differentiation ,Osteoblast ,Articles ,3T3 Cells ,Cell Biology ,Molecular Weight ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Focal Adhesion Protein-Tyrosine Kinases ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Nitric Oxide Synthase ,Protein Tyrosine Phosphatases ,Signal transduction ,Signal Transduction - Abstract
Osteopontin (OPN), a major marker of osteogenic differentiation, suppresses osteoblast responses to mechanical stress and cytokines, including HGF and PDGF. These OPN-induced effects are mediated through focal adhesion kinase inactivation by the induction of low–molecular weight protein tyrosine phosphatase., Osteopontin (OPN) is an osteogenic marker protein. Osteoblast functions are affected by inflammatory cytokines and pathological conditions. OPN is highly expressed in bone lesions such as those in rheumatoid arthritis. However, local regulatory effects of OPN on osteoblasts remain ambiguous. Here we examined how OPN influences osteoblast responses to mechanical stress and growth factors. Expression of NO synthase 1 (Nos1) and Nos2 was increased by low-intensity pulsed ultrasound (LIPUS) in MC3T3-E1 cells and primary osteoblasts. The increase of Nos1/2 expression was abrogated by both exogenous OPN overexpression and recombinant OPN treatment, whereas it was promoted by OPN-specific siRNA and OPN antibody. Moreover, LIPUS-induced phosphorylation of focal adhesion kinase (FAK), a crucial regulator of mechanoresponses, was down-regulated by OPN treatments. OPN also attenuated hepatocyte growth factor–induced vitamin D receptor (Vdr) expression and platelet-derived growth factor–induced cell mobility through the repression of FAK activity. Of note, the expression of low–molecular weight protein tyrosine phosphatase (LMW-PTP), a FAK phosphatase, was increased in both OPN-treated and differentiated osteoblasts. CD44 was a specific OPN receptor for LWW-PTP induction. Consistently, the suppressive influence of OPN on osteoblast responsiveness was abrogated by LMW-PTP knockdown. Taken together, these results reveal novel functions of OPN in osteoblast physiology.
- Published
- 2017
21. Hepatocyte growth factor reduces CXCL10 expression in keratinocytes
- Author
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Kenjiro Bandow, Tetsuya Matsuguchi, Joji Kusuyama, Kyoko Kakimoto, Tomokazu Ohnishi, Takuro Kanekura, and Mitsuhiro Hisadome
- Subjects
Keratinocytes ,Vascular Endothelial Growth Factor A ,0301 basic medicine ,Chemokine ,MAP Kinase Signaling System ,Angiogenesis ,Biophysics ,Dermatology ,Biochemistry ,Cell Line ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Structural Biology ,Genetics ,medicine ,Animals ,Humans ,CXCL10 ,Molecular Biology ,integumentary system ,biology ,Hepatocyte Growth Factor ,Tumor Necrosis Factor-alpha ,Cell Biology ,Chemokine CXCL10 ,Vascular endothelial growth factor ,HaCaT ,Vascular endothelial growth factor A ,030104 developmental biology ,Gene Expression Regulation ,chemistry ,030220 oncology & carcinogenesis ,Immunology ,Cancer research ,biology.protein ,Hepatocyte growth factor ,Wound healing ,medicine.drug - Abstract
Keratinocytes secrete vascular endothelial growth factor (VEGF) and angioregulatory chemokines during cutaneous wound healing. Hepatocyte growth factor (HGF) promotes skin re-epithelialization by increasing VEGF expression in keratinocytes. Here, we investigated the regulatory roles of HGF in the expression of genes encoding angiogenic and angiostatic chemokines in keratinocytes and found that HGF specifically inhibits mRNA expression of the angiostatic chemokine CXCL10 in both mouse primary keratinocytes and in the human keratinocyte cell line HaCaT through the MEK/ERK cascade. Furthermore, HGF inhibited tumor necrosis factor-α-induced CXCL10 expression at both mRNA and protein levels in HaCaT cells. Thus, HGF may orchestrate angiogenesis in wounded skin by modulating both VEGF and CXCL10 expression in keratinocytes.
- Published
- 2016
22. QSAR analysis of tumor-specificity of newly synthesized 3-styrylchromone derivatives against human oral squamous cell carcinoma cell lines
- Author
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Junko Nagai, Hiroshi Sakagami, Koichi Takao, Haixia Shi, Yoshihiro Uesawa, Kenjiro Bandow, Yoshiaki Sugita, Kaori Hoshi, Akito Tomomura, and Mineko Tomomura
- Subjects
Quantitative structure–activity relationship ,Cell culture ,Chemistry ,Applied Mathematics ,General Mathematics ,Cancer research ,Basal cell - Published
- 2020
23. Quantitative Structure-Cytotoxicity Relationship of Pyrano[4,3
- Author
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Junko, Nagai, Haixia, Shi, Yuka, Kubota, Kenjiro, Bandow, Noriyuki, Okudaira, Yoshihiro, Uesawa, Hiroshi, Sakagami, Mineko, Tomomura, Akito, Tomomura, Koichi, Takao, and Yoshiaki, Sugita
- Subjects
Chromones ,Doxorubicin ,Cell Line, Tumor ,Carcinoma, Squamous Cell ,Humans ,Quantitative Structure-Activity Relationship ,Antineoplastic Agents ,Apoptosis ,Female ,Mouth Neoplasms ,Child ,Cell Line - Abstract
4H-1-Benzopyran-4-one (chromone) provides a backbone structure for the chemical synthesis of potent anticancer drugs. Since studies of the biological activity of pyrano[4,3-b]chromones are limited, we investigated a total of 20 pyrano[4,3-b]chromones (10 sets of diastereomers) for their cytotoxicity against four human oral squamous cell carcinoma (OSCC) cell lines and human normal oral cells, and then carried out a quantitative structure-activity relationship (QSAR) analysis.Cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Tumor-specificity (TS) was evaluated by the ratio of mean 50% cytotoxic concentration (CC8-Chloro-4,4a-dihydro-3-methoxy-3-methyl-3H,10H-pyrano[4,3-b][1]benzopyran-10-one (Chemical modification of
- Published
- 2018
24. Quantitative Structure-Cytotoxicity Relationship of 3-(
- Author
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Haixia, Shi, Junko, Nagai, Tsukasa, Sakatsume, Kenjiro, Bandow, Noriyuki, Okudaira, Yoshihiro, Uesawa, Hiroshi, Sakagami, Mineko, Tomomura, Akito, Tomomura, Koichi, Takao, and Yoshiaki, Sugita
- Subjects
Keratinocytes ,Chromones ,Doxorubicin ,Cell Line, Tumor ,Cell Cycle ,Carcinoma, Squamous Cell ,Humans ,Quantitative Structure-Activity Relationship ,Antineoplastic Agents ,Apoptosis ,Mouth Neoplasms ,Cell Line - Abstract
4H-1-Benzopyran-4-ones (chromones) provide a backbone structure for the chemical synthesis of potent anticancer drugs. In contrast to 2-(N-cyclicamino)chromones, the biological activity of 3-(N-cyclicamino)chromones has not been reported. In this study, cytotoxicity of 15 3-(N-cyclicamino)chromone derivatives was investigated and subjected to quantitative structure-activity relationship (QSAR) analysis.Cytotoxicity against four human oral squamous cell carcinoma cell lines and three oral normal mesenchymal cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Tumor-specificity (TS) was evaluated as the ratio of mean 50% cytotoxic concentration (CC3-(4-phenyl-1-piperazinyl)-4H-1-benzopyran-4-one (Chemical modification of
- Published
- 2018
25. Quantitative Structure-Cytotoxicity Relationship of 2-(
- Author
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Haixia, Shi, Junko, Nagai, Tsukasa, Sakatsume, Kenjiro, Bandow, Noriyuki, Okudaira, Hiroshi, Sakagami, Mineko, Tomomura, Akito, Tomomura, Yoshihiro, Uesawa, Koichi, Takao, and Yoshiaki, Sugita
- Subjects
Chromones ,Cell Line, Tumor ,Cell Cycle ,Carcinoma, Squamous Cell ,Humans ,Quantitative Structure-Activity Relationship ,Antineoplastic Agents ,Apoptosis ,Mouth Neoplasms ,Drug Screening Assays, Antitumor - Abstract
4H-1-Benzopyran-4-ones (chromones) have provided backbone structure for the chemical synthesis of potent anticancer drugs. In this study, the cytotoxicity of fifteen 2-(N-cyclicamino)chromone derivatives was investigated and subjected to quantitative structure-activity relationship (QSAR) analysis.Cytotoxicity against four human oral squamous cell carcinoma cell lines and three oral normal mesenchymal cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Tumor specificity (TS) was evaluated by ratio of mean 50% cytotoxic concentration (CC7-Methoxy-2-(4-morpholinyl)-4H-1-benzopyran-4-one (Chemical modification of
- Published
- 2018
26. Constitutive activation of p46JNK2 is indispensable for C/EBPδ induction in the initial stage of adipogenic differentiation
- Author
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Kaori Shima, Kenjiro Bandow, Muhammad Subhan Amir, Joji Kusuyama, Tetsuya Matsuguchi, Tomokazu Ohnishi, and Ichiro Semba
- Subjects
0301 basic medicine ,CCAAT-Enhancer-Binding Protein-delta ,medicine.medical_specialty ,Biochemistry ,03 medical and health sciences ,Mice ,Downregulation and upregulation ,Internal medicine ,3T3-L1 Cells ,medicine ,Adipocytes ,Animals ,Mitogen-Activated Protein Kinase 9 ,Molecular Biology ,Transcription factor ,Anthracenes ,Adipogenesis ,biology ,Dose-Response Relationship, Drug ,Kinase ,Promoter ,Cell Differentiation ,Cell Biology ,Activating transcription factor 2 ,Cell biology ,Intracellular signal transduction ,Enzyme Activation ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,biology.protein ,Phosphorylation - Abstract
Adipogenic differentiation plays a vital role in energy homeostasis and endocrine system. Several transcription factors including peroxisome proliferator-activated receptor γ 2 (PPARγ2), CCAAT-enhancer-binding protein (C/EBP) α, β, and δ are important for the process, whereas the stage-specific intracellular signal transduction regulating the onset of adipogenesis remains enigmatic. Here, we explored the functional role of c- jun N-terminal kinases (JNKs) in adipogenic differentiation using in vitro differentiation models of 3T3-L1 cells and primary adipo-progenitor cells. JNK inactivation with either a pharmacological inhibitor or JNK2-specific siRNA suppressed adipogenic differentiation, characterized by decreased lipid droplet appearance and the downregulation of Adiponectin , fatty acid protein 4 ( Fabp4 ), Pparg2 , and C/ebpa expressions. Conversely, increased adipogenesis was observed by the inducible overexpression of p46JNK2 (JNK2-1), whereas it was not observed by that of p54JNK2 (JNK2-2), indicating a distinct role of p46JNK2. The essential role of JNK appears restricted to the early stage of adipogenic differentiation, as JNK inhibition in the later stages did not influence adipogenesis. Indeed, JNK phosphorylation was significantly induced at the onset of adipogenic differentiation. As for the transcription factors involved in early adipogenesis, JNK inactivation significantly inhibited the induction of C/ebpd , but not C/ebpb, during the initial stage of adiogenic differentiation. JNK activation increased C/ebpd mRNA and protein expression through the induction and phosphorylation of activating transcription factor 2 (ATF2) that binds to a responsive element within the C/ebpd gene promoter region. Taken together, these data indicate that constitutive JNK activity is specifically required for the initial stage differentiation events of adipocytes.
- Published
- 2017
27. Low-intensity pulsed ultrasound (LIPUS) inhibits LPS-induced inflammatory responses of osteoblasts through TLR4–MyD88 dissociation
- Author
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Kyoko Kakimoto, Yasuyuki Fujii, Kenjiro Bandow, Tetsuya Matsuguchi, Joji Kusuyama, Juna Nakao, and Tomokazu Ohnishi
- Subjects
Lipopolysaccharides ,Transcriptional Activation ,musculoskeletal diseases ,medicine.medical_specialty ,Histology ,Physiology ,Ultrasonic Therapy ,Endocrinology, Diabetes and Metabolism ,p38 mitogen-activated protein kinases ,Lymphocyte Antigen 96 ,Low-intensity pulsed ultrasound ,CCL2 ,Models, Biological ,Cell Line ,Mice ,Internal medicine ,medicine ,Animals ,Ultrasonics ,RNA, Messenger ,Protein kinase B ,Inflammation ,Osteoblasts ,Kinase ,Chemistry ,Gene Expression Profiling ,Cell Membrane ,RANK Ligand ,NF-kappa B ,Osteoblast ,Cell biology ,Mice, Inbred C57BL ,Toll-Like Receptor 4 ,Endocrinology ,medicine.anatomical_structure ,Myeloid Differentiation Factor 88 ,TLR4 ,lipids (amino acids, peptides, and proteins) ,Chemokines ,Signal transduction ,Signal Transduction - Abstract
Previous reports have shown that osteoblasts are mechano-sensitive. Low-intensity pulsed ultrasound (LIPUS) induces osteoblast differentiation and is an established therapy for bone fracture. Here we have examined how LIPUS affects inflammatory responses of osteoblasts to LPS. LPS rapidly induced mRNA expression of several chemokines including CCL2, CXCL1, and CXCL10 in both mouse osteoblast cell line and calvaria-derived osteoblasts. Simultaneous treatment by LIPUS significantly inhibited mRNA induction of CXCL1 and CXCL10 by LPS. LPS-induced phosphorylation of ERKs, p38 kinases, MEK1/2, MKK3/6, IKKs, TBK1, and Akt was decreased in LIPUS-treated osteoblasts. Furthermore, LIPUS inhibited the transcriptional activation of NF-κB responsive element and Interferon-sensitive response element (ISRE) by LPS. In a transient transfection experiment, LIPUS significantly inhibited TLR4-MyD88 complex formation. Thus LIPUS exerts anti-inflammatory effects on LPS-stimulated osteoblasts by inhibiting TLR4 signal transduction.
- Published
- 2014
28. Low intensity pulsed ultrasound (LIPUS) maintains osteogenic potency by the increased expression and stability of Nanog through spleen tyrosine kinase (Syk) activation
- Author
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Kenjiro Bandow, Ichiro Semba, Tetsuya Matsuguchi, Nathan S. Makarewicz, Joji Kusuyama, Kaori Shima, Tomokazu Ohnishi, and Changhwan Seong
- Subjects
0301 basic medicine ,Homeobox protein NANOG ,Stromal cell ,Syk ,Mice ,03 medical and health sciences ,Paracrine signalling ,0302 clinical medicine ,SOX2 ,Osteogenesis ,medicine ,Animals ,Syk Kinase ,rho-Associated Kinases ,Osteoblasts ,biology ,Chemistry ,SOXB1 Transcription Factors ,Mesenchymal stem cell ,Gene Expression Regulation, Developmental ,Cell Differentiation ,Mesenchymal Stem Cells ,Osteoblast ,Nanog Homeobox Protein ,Cell Biology ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Ultrasonic Waves ,030220 oncology & carcinogenesis ,embryonic structures ,Osteocalcin ,biology.protein ,biological phenomena, cell phenomena, and immunity - Abstract
Mesenchymal stem cells (MSCs) are a powerful tool for cell-based, clinical therapies like bone regeneration. Therapeutic use of cell transplantation requires many cells, however, the expansion process needed to produce large quantities of cells reduces the differentiation potential of MSCs. Here, we examined the protective effects of low intensity pulsed ultrasound (LIPUS) on the maintenance of osteogenic potency. Primary osteoblastic cells were serially passaged between 2 and 12 times with daily LIPUS treatment. We found that LIPUS stimulation maintains osteogenic differentiation capacity in serially passaged cells, as characterized by improved matrix mineralization and Osteocalcin mRNA expression. Decreased expression of Nanog, Sox2, and Msx2, and increased expression of Pparg2 from serial passaging was recovered in LIPUS-stimulated cells. We found that LIPUS stimulation not only increased but also sustained expression of Nanog in primary osteoblasts and ST2 cells, a mouse mesenchymal stromal cell line. Nanog overexpression in serially passaged cells mimicked the recuperative effects of LIPUS on osteogenic potency, highlighting the important role of Nanog in LIPUS stimulation. Additionally, we found that spleen tyrosine kinase (Syk) is an important signaling molecule to induce Nanog expression in LIPUS-stimulated cells. Syk activation was regulated by both Rho-associated kinase 1 (ROCK1) and extracellular ATP in a paracrine manner. Interestingly, the LIPUS-induced increase in Nanog mRNA expression was regulated by ATP-P2X4-Syk Y323 activation, while the improvement of Nanog protein stability was controlled by the ROCK1-Syk Y525/526 pathway. Taken together, these results indicate that LIPUS stimulation recovers and maintains the osteogenic potency of serially passaged cells through a Syk-Nanog axis.
- Published
- 2019
29. Synthesis and biological evaluation of some new mono Mannich bases with piperazines as possible anticancer agents and carbonic anhydrase inhibitors
- Author
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Claudiu T. Supuran, Yusuf Özkay, Halise Inci Gul, Kenjiro Bandow, Mehtap Tugrak, Hiroshi Sakagami, İlhami Gülçin, Anadolu Üniversitesi, Eczacılık Fakültesi, Farmasötik Kimya Anabilim Dalı, and Özkay, Yusuf
- Subjects
Chalcone ,Carbonic Anhydrase I ,Cytotoxicity ,Antineoplastic Agents ,Apoptosis ,Carbonic Anhydrase II ,01 natural sciences ,Biochemistry ,Piperazines ,Carbonic Anhydrase ,Mannich Bases ,Structure-Activity Relationship ,chemistry.chemical_compound ,Neoplasms ,Carbonic anhydrase ,Drug Discovery ,Tumor Cells, Cultured ,Humans ,Phenol ,Carbonic Anhydrase Inhibitors ,Molecular Biology ,Cell Proliferation ,Biological evaluation ,Molecular Structure ,biology ,010405 organic chemistry ,Organic Chemistry ,Combinatorial chemistry ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,chemistry ,biology.protein - Abstract
4th Symposium on Biotransformations for Pharmaceutical and Cosmetic Industry -- JUN 25-27, 2018 -- Trzebnica, POLAND, WOS: 000479184600049, PubMed ID: 31288135, New mono Mannich bases, (2-(4-hydroxy-3-((4-substituephenylpiperazin-1-yl)methyl)benzylidene)-2,3-dihydro-1H-inden-1-one), were prepared to evaluate their cytotoxic/anticancer properties and also their inhibitory effects on human carbonic anhydrase I and II isoenzymes (hCA I and II). Amine part was changed as [N-phenylpiperazine (1),N-benzylpiperazine (2), 1-(2-fluorophenyl)piperazine (3), 1-(4-fluorophenyl)piperazine (4), 1-(2-methoxyphenyl)piperazine (5)]. The structure of the synthesized compounds was characterized by H-1 NMR, C-13 NMR and HRMS spectra. Cytotoxicity results of the series pointed out that the compound 4 had the highest tumor selectivity value (TS: 59.4) possibly by inducing necrotic cell death in series. Additionally, all compounds synthesized showed a good inhibition profile towards hCA I and II isoenzymes with the Ki values between 29.6 and 58.4 nM and 38.1-69.7 nM, respectively. These values were lower than the reference compound AZA. However, it seems that the compounds 4 and 2 can be considered as lead compounds of CA studies with the lowest Ki values in series for further designs., Ataturk University BAP Office, The authors are thankful to Dr. C. Kazaz and Dr. B. Anil (Ataturk University) for NMRs and Ataturk University BAP Office for their financial support.
- Published
- 2019
30. 10. Low-Intensity Pulsed Ultrasound (LIPUS) Stimulation Helps to Maintain the Differentiation Potency of Mesenchymal Stem Cells by Induction in Nanog Protein Transcript Levels and Phosphorylation
- Author
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Joji Kusuyama, Chang Hwan Seong, Tomokazu Ohnishi, Tetsuya Matsuguchi, and Kenjiro Bandow
- Subjects
0301 basic medicine ,Homeobox protein NANOG ,Mesenchymal stem cell ,General Medicine ,Biology ,Low-intensity pulsed ultrasound ,Stem cell marker ,Molecular biology ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Cell culture ,Myocyte ,Phosphorylation ,Orthopedics and Sports Medicine ,Surgery ,Subculture (biology) - Abstract
Mesenchymal Stem Cells (MSCs) are pluripotent cells that can be differentiated as osteoblasts, adipocytes, myocytes or chondrocytes depending on the culture condition. However, MSCs are known to lose their differentiation potency after long-term culture. Development of a new cell culture method to maintain their stemness is required for successful application of MSCs. Here, we revealed that low-intensity pulsed ultrasound (LIPUS) stimulation was useful for maintaining the MSC stemness as LIPUS inhibited the loss of osteogenic differentiation potency of osteo-progenitor cells induced by serial subculture. LIPUS also increased the transcriptional and phosphorylation level of Nanog, a crucial stem cell marker gene in a MSC cell line. We also found that LIPUS induced the secretion of extracellular adenosine triphosphate (ATP) in MSC. The treatments of the conditioned medium from LIPUS-stimulated MSC and exogenous ATP promoted Nanog expression. Thus, LIPUS may maintain the long-term differentiation potency of MSCs and osteo-progenitor cells by induction in Nanog transcript level and phosphorylation.
- Published
- 2016
31. LPS-induced chemokine expression in both MyD88-dependent and -independent manners is regulated by Cot/Tpl2-ERK axis in macrophages
- Author
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Kenjiro Bandow, Tetsuya Matsuguchi, Mitsuo Shamoto, Joji Kusuyama, Tomokazu Ohnishi, and Kyoko Kakimoto
- Subjects
Lipopolysaccharides ,LPS ,Macrophage ,Biophysics ,CCL7 ,Biochemistry ,Cell Line ,Mice ,Cot/Tpl2 ,Structural Biology ,Genetics ,Animals ,CXCL10 ,Extracellular Signal-Regulated MAP Kinases ,Molecular Biology ,DNA Primers ,Base Sequence ,biology ,Chemistry ,Macrophages ,Cell Biology ,Molecular biology ,Mice, Inbred C57BL ,ERK ,CXCL2 ,CXCL3 ,Chemokine ,TRIF ,Mitogen-activated protein kinase ,Myeloid Differentiation Factor 88 ,biology.protein ,CXCL9 ,Chemokines ,Signal transduction - Abstract
LPS signaling is mediated through MyD88-dependent and -independent pathways, activating NF-κB, MAP kinases and IRF3. Cot/Tpl2 is an essential upstream kinase in LPS-mediated activation of ERKs. Here we explore the roles of MyD88 and Cot/Tpl2 in LPS-induced chemokine expression by studying myd88−/− and cot/tpl2−/− macrophages. Among the nine LPS-responsive chemokines examined, mRNA induction of ccl5, cxcl10, and cxcl13 is mediated through the MyD88-independent pathway. Notably, Cot/Tpl2-ERK signaling axis exerts negative effects on the expression of these three chemokines. In contrast, LPS-induced gene expression of ccl2, ccl7, cxcl2, cxcl3, ccl8, and cxcl9 is mediated in the MyD88-dependent manner. The Cot/Tpl2-ERK axis promotes the expression of the first four and inhibits the expression of the latter two. Thus, LPS induces expression of multiple chemokines through various signaling pathways in macrophages.
- Published
- 2012
32. Search of Neuroprotective Polyphenols Using the 'Overlay' Isolation Method
- Author
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Kenjiro Bandow, Misaki Horiuchi, Hiroshi Sakagami, Akito Tomomura, Haixia Shi, Takaaki Oizumi, Mineko Tomomura, and Tomohiro Fujisawa
- Subjects
Cell Culture Techniques ,Pharmaceutical Science ,Epigallocatechin gallate ,Resveratrol ,Pharmacology ,PC12 Cells ,Catechin ,Culture Media, Serum-Free ,overlay method ,Analytical Chemistry ,chemistry.chemical_compound ,0302 clinical medicine ,Blood serum ,hormesis ,Nerve Growth Factor ,Stilbenes ,Drug Discovery ,bamboo leaf extract ,Cytotoxic T cell ,Neurons ,NGF ,Cell Differentiation ,PC12 ,differentiation ,taxanes ,Neuroprotective Agents ,Chemistry (miscellaneous) ,030220 oncology & carcinogenesis ,Molecular Medicine ,Taxoids ,neuroprotection ,Sasa ,Curcumin ,amyloid-β peptide ,Neuroprotection ,Article ,lcsh:QD241-441 ,03 medical and health sciences ,lcsh:Organic chemistry ,Animals ,Physical and Theoretical Chemistry ,Amyloid beta-Peptides ,Plant Extracts ,Organic Chemistry ,Polyphenols ,Rats ,Plant Leaves ,polyphenol ,Nerve growth factor ,chemistry ,Polyphenol ,030217 neurology & neurosurgery - Abstract
Previous studies of the neuroprotective activity of polyphenols have used ununiform culture systems, making it difficult to compare their neuroprotective potency. We have established a new and simple method for preparing differentiated PC12 cells by removing the toxic coating step. Cells were induced to differentiate with the nerve growth factor (NGF) in a serum-free medium, without a medium change, but with a one-time overlay supplementation of NGF. The optimal inoculation density of the cells was 6&ndash, 12 ×, 103 cells/cm2, and the presence of serum inhibited the differentiation. Neuroprotective activity could be quantified by the specific index (SI) value, that is, the ratio of the 50% cytotoxic concentration to the 50% effective concentration. Alkaline extract from the leaves of Sasa senanensis Rehder (SE), having had hormetic growth stimulation, showed the highest SI value, followed by epigallocatechin gallate. The SI value of curcumin and resveratrol was much lower. This simple overly method, that can prepare massive differentiated neuronal cells, may be applicable for the study of the differentiation-associated changes in intracellular metabolites, and the interaction between neuronal cells and physiological factors.
- Published
- 2018
33. Cot/Tpl2 regulates IL-23 p19 expression in LPS-stimulated macrophages through ERK activation
- Author
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Tipayaratn Musikacharoen, Kenjiro Bandow, Tetsuya Matsuguchi, Kyoko Kakimoto, Norika Chiba, and Tomokazu Ohnishi
- Subjects
Lipopolysaccharides ,MAPK/ERK pathway ,Lipopolysaccharide ,Physiology ,Mice, Transgenic ,Biology ,Biochemistry ,Mice ,chemistry.chemical_compound ,Proto-Oncogene Proteins ,Extracellular ,Animals ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Protein kinase A ,Receptor ,Antigen-presenting cell ,Kinase ,Macrophages ,Cell Differentiation ,General Medicine ,MAP Kinase Kinase Kinases ,Interleukin-12 ,Cell biology ,Mice, Inbred C57BL ,chemistry ,Interleukin-23 Subunit p19 ,Cancer research ,TLR4 - Abstract
We have previously reported that a serine/threonine protein kinase, Cot/Tpl2, is a negative regulator of Th1-type immunity through inhibiting IL-12 expression in antigen presenting cells (APCs) stimulated by Toll-like receptor (TLR) ligands. We here show that Cot/Tpl2(-/-) macrophages produce significantly less IL-23, an important regulator of Th17-type response, than the wild-type counterparts in response to lipopolysaccharide (LPS), which is a ligand for TLR4. The decreased IL-23 production in Cot/Tpl2(-/-) macrophages is, at least partly, regulated at the transcriptional level, as the LPS-mediated IL-23 p19 mRNA induction was significantly less in Cot/Tpl2(-/-) macrophages. Chemical inhibition of extracellular signal-regulated kinase (ERK) activity similarly inhibited IL-23 expression in LPS-stimulated wild-type macrophages. As Cot/Tpl2 is an essential upstream component of the ERK activation pathway of LPS, it is suggested that Cot/Tpl2 positively regulates IL-23 expression through ERK activation. These results indicate that Cot/Tpl2 may be involved in balancing Th1/Th17 differentiation by regulating the expression ratio of IL-12 and IL-23 in APCs.
- Published
- 2010
34. Involvement of Cot/Tp12 in Bone Loss during Periodontitis
- Author
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Tomokazu Ohnishi, Tetsuya Matsuguchi, A. Okamoto, Kyoko Kakimoto, Kenjiro Bandow, and Norika Chiba
- Subjects
Lipopolysaccharides ,Male ,medicine.medical_specialty ,Lipopolysaccharide ,MAP Kinase Signaling System ,Periodontal Ligament ,Alveolar Bone Loss ,Osteoclasts ,Mice ,chemistry.chemical_compound ,Proto-Oncogene Proteins ,Internal medicine ,medicine ,Animals ,Periodontitis ,Receptor ,General Dentistry ,Dental alveolus ,Mice, Knockout ,Mitogen-Activated Protein Kinase Kinases ,Osteoblasts ,biology ,Tumor Necrosis Factor-alpha ,business.industry ,RANK Ligand ,Cell Differentiation ,MAP Kinase Kinase Kinases ,medicine.disease ,Pathophysiology ,Resorption ,Mice, Inbred C57BL ,Osteopenia ,Endocrinology ,chemistry ,Cyclooxygenase 2 ,RANKL ,biology.protein ,Cytokines ,business - Abstract
Periodontitis causes resorption of alveolar bone, in which RANKL induces osteoclastogenesis. The binding of lipopolysaccharide to Toll-like receptors causes phosphorylation of Cot/Tp12 to activate the MAPK cascade. Previous in vitro studies showed that Cot/Tp12 was essential for the induction of RANKL expression by lipopolysaccharide. In this study, we examined whether Cot/Tp12 deficiency reduced the progression of alveolar bone loss and osteoclastogenesis during experimental periodontitis. We found that the extent of alveolar bone loss and osteoclastogenesis induced by ligature-induced periodontitis was decreased in Cot/Tp12-deficient mice. In addition, reduction of RANKL expression was observed in periodontal tissues of Cot/Tp12-deficient mice with experimental periodontitis. Furthermore, we found that Cot/Tp12 was involved in the induction of TNF-α mRNA expression in gingiva of mice with experimental periodontitis. Our observations suggested that Cot/Tp12 is essential for the progression of alveolar bone loss and osteoclastogenesis in periodontal tissue during experimental periodontitis mediated through increased RANKL expression.
- Published
- 2010
35. Reduction of orthodontic tooth movement by experimentally induced periodontal inflammation in mice
- Author
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Atsuko Okamoto, Tomohiro Fukunaga, Tomokazu Ohnishi, Kyoko Kakimoto, Tetsuya Matsuguchi, Norika Chiba, Shouichi Miyawaki, Kenjiro Bandow, and Aya Maeda
- Subjects
Male ,Tooth Movement Techniques ,medicine.medical_treatment ,Acid Phosphatase ,Alveolar Bone Loss ,Osteoclasts ,Dentistry ,Mice, Inbred Strains ,Oral hygiene ,Dinoprostone ,Mice ,Osteoclast ,Alveolar Process ,Maxilla ,Animals ,Medicine ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Periodontitis ,Ligature ,General Dentistry ,Dental alveolus ,Tartrate-resistant acid phosphatase ,Osteoblasts ,biology ,Tartrate-Resistant Acid Phosphatase ,business.industry ,RANK Ligand ,3T3 Cells ,medicine.disease ,Molar ,Isoenzymes ,Mice, Inbred C57BL ,Disease Models, Animal ,medicine.anatomical_structure ,Cyclooxygenase 2 ,RANKL ,biology.protein ,Stress, Mechanical ,business ,Biomarkers ,Prostaglandin E - Abstract
Orthodontic therapy is known to have an aggravating effect on the progression of destructive periodontitis if oral hygiene is not maintained. However, it is largely unknown how active periodontitis affects the velocity of orthodontic tooth movement. In this study, we examined the effect of periodontal inflammation on orthodontic tooth movement using a mouse model. Orthodontic force was applied on the maxillary first molar of mice, with or without ligature wire to induce experimental periodontitis. The distance moved by the first molar was significantly reduced by the ligature-induced experimental periodontitis. Tartrate-resistant acid phosphatase staining revealed that the number of osteoclasts present during orthodontic treatment was lower in the pressure zone of alveolar bone in the presence of periodontal inflammation. Consistently, the expression level of receptor activator of nuclear factor-kappaB ligand (RANKL) in the pressure zone was decreased in the ligature group. By contrast, experimental periodontitis increased the expression of cyclooxygenase-2 mRNA in the periodontal tissues, while in vitro treatment with prostaglandin E(2) decreased extracellular signal-regulated kinase phosphorylation and RANKL expression induced by mechanical stress in osteoblasts. Taken together, these results suggest that the orthodontic force-induced osteoclastogenesis in alveolar bone was inhibited by the accompanying periodontal inflammation, at least partly through prostaglandin E(2), resulting in reduced orthodontic tooth movement.
- Published
- 2009
36. Oxidative stress causes alveolar bone loss in metabolic syndrome model mice with type 2 diabetes
- Author
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Tomokazu Ohnishi, Miho Machigashira, Kyoko Kakimoto, Tetsuya Matsuguchi, Takashi Matsuyama, and Kenjiro Bandow
- Subjects
Keratinocytes ,Male ,medicine.medical_specialty ,Antioxidant ,Nitric Oxide Synthase Type III ,medicine.medical_treatment ,Alveolar Bone Loss ,Gingiva ,Mice, Inbred Strains ,Stimulation ,Mandible ,medicine.disease_cause ,Antioxidants ,Mice ,chemistry.chemical_compound ,Internal medicine ,medicine ,Animals ,Enzyme Inhibitors ,Hydrogen peroxide ,Cells, Cultured ,Dental alveolus ,Metabolic Syndrome ,chemistry.chemical_classification ,Periodontitis ,Reactive oxygen species ,Insulin ,Free Radical Scavengers ,Hydrogen Peroxide ,medicine.disease ,Acetylcysteine ,Mice, Inbred C57BL ,Disease Models, Animal ,Oxidative Stress ,Endocrinology ,Diabetes Mellitus, Type 2 ,chemistry ,Periodontics ,Reactive Oxygen Species ,Oxidative stress - Abstract
Background and Objective: Alveolar bone loss is caused by a host response to periodontal pathogens, and its progression is often enhanced by systemic conditions such as insulin resistance.Alveolar bone dehiscence has been observed in KK-Ay mice, which are metabolic syndrome model mice with type 2 diabetes. The aim of this study was to investigate inducements responsible for alveolar bone dehiscence in the KK-Ay mice. Material and Methods: The expression of endothelial nitric oxide synthase in the mandibles of mice was detected using immunohistochemical staining and the reverse transcription–polymerase chain reaction. After administration of N-acetylcysteine, an antioxidant, to KK-Ay mice, alveolar bone loss and the expression of endothelial nitric oxide synthase protein in gingival keratinocytes and of hydrogen peroxide concentrations in plasma, were analyzed. The effect of hydrogen peroxide on endothelial nitric oxide synthase expression in keratinocytes was examined using cultured keratinocytes. Results: The expression of endothelial nitric oxide synthase was decreased in gingival keratinocytes from KK-Ay mice compared with gingival keratinocytes from control mice. Administration of N-acetylcysteine to the mice restored endothelial nitric oxide synthase expression in the gingival keratinocytes, suppressed the alveolar bone loss and decreased the hydrogen peroxide concentrations in plasma without the improvement of obesity or diabetes. In vitro, stimulation with hydrogen peroxide decreased the expression level of endothelial nitric oxide synthase in cultured keratinocytes, which was restored by the addition of N-acetylcysteine. Conclusion: Reactive oxygen species, such as hydrogen peroxide, are responsible for the alveolar bone loss accompanied by decreased endothelial nitric oxide synthase expression in KK-Ay mice. Therefore, we propose a working hypothesis that the generation of oxidative stress is an underlying systemic condition that enhances alveolar bone loss in periodontitis occurring as a complication of diabetes.
- Published
- 2009
37. Dehydroepiandrosterone increased oxidative stress in a human cell line during differentiation
- Author
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Kohji Aoyama, Kimiko Izumo, Tetsuya Matsuguchi, Masaharu Komatsu, Minoru Takeuchi, Toru Takeuchi, Masahisa Horiuchi, and Kenjiro Bandow
- Subjects
endocrine system ,medicine.medical_specialty ,Antioxidant ,Cell Survival ,HL60 ,medicine.medical_treatment ,Dehydroepiandrosterone ,HL-60 Cells ,Stimulation ,Cell Growth Processes ,Glucosephosphate Dehydrogenase ,medicine.disease_cause ,Biochemistry ,chemistry.chemical_compound ,Internal medicine ,polycyclic compounds ,medicine ,Humans ,Glucose-6-phosphate dehydrogenase ,Dimethyl Sulfoxide ,RNA, Messenger ,skin and connective tissue diseases ,Chemistry ,Superoxide ,NADPH Oxidases ,Cell Differentiation ,General Medicine ,Glutathione ,Oxidative Stress ,Endocrinology ,Reactive Oxygen Species ,human activities ,hormones, hormone substitutes, and hormone antagonists ,Oxidative stress - Abstract
Dehydroepiandrosterone (DHEA), a reversible inhibitor of glucose-6-phosphate dehydrogenase (G6PD), is increasingly taken as an antioxidative and anti-ageing supplement. This study investigated the effects of DHEA on the expression of G6PD and on the state of oxidative stress in a human promyelocytic leukaemia cell line, HL60, during the differentiation to neutrophil-like cell. This study differentiated HL60 with dimethyl sulfoxide (DMSO) in the presence (DMSO-HL60/DHEA) or absence (DMSO-HL60) of DHEA. During the differentiation, activity, mRNA and protein levels of G6PD were increased. DHEA increased these levels further. DHEA by itself suppressed the production of superoxide from DMSO-HL60 upon stimulation with phorbol myristate acetate (PMA). However, DMSO-HL60/DHEA stimulated with PMA in the absence of DHEA produced superoxide and 8-oxo-deoxyguanosine more than PMA-stimulated DMSO-HL60. After addition of H(2)O(2), the ratio of reduced glutathione to oxidized glutathione was lower in DMSO-HL60/DHEA than in DMSO-HL60. These findings indicate that DHEA acts both as an antioxidant and as a pro-oxidant.
- Published
- 2009
38. Hepatocyte growth factor/scatter factor stimulates migration of muscle precursors in developing mouse tongue
- Author
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Ichiro Semba, Yasushi Daikuhara, Masato Tamura, Kenjiro Bandow, and Tomokazu Ohnishi
- Subjects
medicine.medical_specialty ,Physiology ,Myoblasts, Skeletal ,Cellular differentiation ,Blotting, Western ,Clinical Biochemistry ,Basic fibroblast growth factor ,Fluorescent Antibody Technique ,Mice ,Phosphatidylinositol 3-Kinases ,chemistry.chemical_compound ,Tongue ,Cell Movement ,Internal medicine ,medicine ,Animals ,Myocyte ,RNA, Messenger ,Muscle, Skeletal ,Myogenin ,Hepatocyte Growth Factor ,Reverse Transcriptase Polymerase Chain Reaction ,Myogenesis ,Skeletal muscle ,Cell Differentiation ,Cell Biology ,Blotting, Northern ,Embryo, Mammalian ,Cell biology ,Enzyme Activation ,Endocrinology ,medicine.anatomical_structure ,Matrix Metalloproteinase 9 ,chemistry ,Fibroblast Growth Factor 2 ,Hepatocyte growth factor ,C2C12 ,Signal Transduction ,medicine.drug - Abstract
Hepatocyte growth factor (HGF) stimulates the migration of myogenic cells during the development of skeletal muscles. The inactivation of HGF genes or that of its receptor, c-met, in mice causes hypoplasia of skeletal muscle organs, such as the tongue. Basic fibroblast growth factor (FGF-2) also induces migration of skeletal myoblasts. A comparison of the functions of HGF and FGF-2 in myogenesis revealed the crucial effect of HGF in the development of skeletal muscles. Unlike FGF-2, HGF induced migration of myoblasts from the developing mouse tongue. The differences between the activities of HGF and FGF-2 were determined by comparing their effects on the expression of matrix metalloproteinase-9 (MMP-9) in myoblasts, C2C12 cells, cultured in collagen-coated dishes. The results showed that HGF, but not FGF-2, stimulated MMP-9 expression, and that the stimulation was mediated through the activation of phosphoinositide 3-kinase (PI3K) which was not associated with FGF-2 signal transduction. Nevertheless, both growth factors exerted almost the same effect on the reduction of myogenin expression in, and on the proliferation of, C2C12 cells, suggesting that HGF, rather than FGF-2, plays a crucial role in the generation of skeletal muscles, including the tongue. Moreover, the specific role of HGF through the PI3K signal pathway is the induction of MMP-9 expression in, and the migration of, myoblasts.
- Published
- 2004
39. Induction of CXCL2 and CCL2 by pressure force requires IL-1β-MyD88 axis in osteoblasts
- Author
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Tetsuya Matsuguchi, Kyoko Kakimoto, Kenjiro Bandow, Joji Kusuyama, Shouichi Miyawaki, Aya Maeda, and Tomokazu Ohnishi
- Subjects
MAPK/ERK pathway ,Chemokine ,Histology ,Physiology ,Endocrinology, Diabetes and Metabolism ,p38 mitogen-activated protein kinases ,Chemokine CXCL2 ,Interleukin-1beta ,CCL2 ,Real-Time Polymerase Chain Reaction ,Bone resorption ,Bone remodeling ,Neutralization Tests ,Pressure ,Animals ,RNA, Messenger ,Cells, Cultured ,Chemokine CCL2 ,Osteoblasts ,biology ,Chemistry ,Cell biology ,Enzyme Activation ,Mice, Inbred C57BL ,CXCL2 ,Mitogen-activated protein kinase ,Myeloid Differentiation Factor 88 ,biology.protein ,Cancer research ,Mitogen-Activated Protein Kinases ,Signal Transduction - Abstract
Mechanical stresses including pressure force induce chemokine expressions in osteoblasts resulting in inflammatory reactions and bone remodeling. However, it has not been well elucidated how mechanical stresses induce inflammatory chemokine expressions in osteoblasts. IL-1β has been identified as an important pathogenic factor in bone loss diseases, such as inflammatory arthritis and periodontitis. Myeloid differentiation factor 88 (MyD88) is an essential downstream adaptor molecule of IL-1 receptor signaling. This study was to examine the gene expression profiles of inflammatory chemokines and the role of MyD88 in osteoblasts stimulated by pressure force. Pressure force (10g/cm(2)) induced significant mRNA increases of CXCL2, CCL2, and CCL5, as well as prompt phosphorylation of MAP kinases (ERK, p38 and JNK), in wild-type primary osteoblasts. The CXCL2 and CCL2 mRNA increases and MAP kinase phosphorylation were severely impaired in MyD88(-/-) osteoblasts. Constitutive low-level expression of IL-1β mRNA was similarly observed in both wild-type and MyD88(-/-) osteoblasts, which was not altered by pressure force stimulation. Notably, neutralization of IL-1β with a specific antibody significantly impaired pressure force-induced mRNA increases of CXCL2 and CCL2, as well as MAP kinase phosphorylation, in wild-type osteoblasts. Furthermore, pre-treatment with recombinant IL-1β significantly enhanced MAP kinase phosphorylation and mRNA increases of CXCL2 and CCL2 by pressure force in wild-type but not MyD88(-/-) osteoblasts. These results have suggested that the activation of MyD88 pathway by constitutive low-level IL-1β expression is essential for pressure force-induced CXCL2 and CCL2 expression in osteoblasts. Thus MyD88 signal in osteoblasts may be required for bone resorption by pressure force through chemokine induction.
- Published
- 2014
40. AMP-activated protein kinase (AMPK) activity negatively regulates chondrogenic differentiation
- Author
-
Joji Kusuyama, Kenjiro Bandow, Tetsuya Matsuguchi, Kyoko Kakimoto, and Tomokazu Ohnishi
- Subjects
Transcriptional Activation ,Histology ,Physiology ,Endocrinology, Diabetes and Metabolism ,Molecular Sequence Data ,AMP-Activated Protein Kinases ,Chondrocyte ,Sodium Selenite ,AMP-activated protein kinase ,Genes, Reporter ,medicine ,Animals ,Humans ,Phosphorylation ,RNA, Small Interfering ,Protein kinase A ,Luciferases ,Aggrecan ,Cells, Cultured ,G alpha subunit ,biology ,Base Sequence ,Chemistry ,Activator (genetics) ,Kinase ,Stem Cells ,Transferrin ,AMPK ,Cell Differentiation ,Extremities ,SOX9 Transcription Factor ,Ribonucleotides ,Aminoimidazole Carboxamide ,Embryo, Mammalian ,Metformin ,Cell biology ,Mice, Inbred C57BL ,Protein Subunits ,medicine.anatomical_structure ,Glucose ,Gene Knockdown Techniques ,biology.protein ,Cattle ,Chondrogenesis - Abstract
Chondrocytes are derived from mesenchymal stem cells, and play an important role in cartilage formation. Sex determining region Y box (Sox) family transcription factors are essential for chondrogenic differentiation, whereas the intracellular signal pathways of Sox activation have not been clearly elucidated. AMP-activated protein kinase (AMPK) is a serine-threonine kinase generally regarded as a key regulator of cellular energy homeostasis. It is known that the catalytic alpha subunit of AMPK is activated by upstream AMPK kinases (AMPKKs) including liver kinase B1 (LKB1). We have previously reported that AMPK is a negative regulator of osteoblastic differentiation. Here, we have explored the role of AMPK in chondrogenic differentiation using in vitro culture models. The phosphorylation level of the catalytic AMPK alpha subunit significantly decreased during chondrogenic differentiation of primary chondrocyte precursors as well as ATDC-5, a well-characterized chondrogenic cell line. Treatment with metformin, an activator of AMPK, significantly reduced cartilage matrix formation and inhibited gene expression of sox6, sox9, col2a1 and aggrecan core protein (acp). Thus, chondrocyte differentiation is functionally associated with decreased AMPK activity.
- Published
- 2014
41. Functional involvement of dual specificity phosphatase 16 (DUSP16), a c-Jun N-terminal kinase-specific phosphatase, in the regulation of T helper cell differentiation
- Author
-
Kenjiro Bandow, Joji Kusuyama, Yasunobu Yoshikai, Tetsuya Matsuguchi, Tipayaratn Musikacharoen, Onishi Tomokazu, and Kyoko Kakimoto
- Subjects
Lymphocyte ,Immunology ,Mice, Transgenic ,GATA3 Transcription Factor ,Biochemistry ,Gene Expression Regulation, Enzymologic ,Histones ,Interleukin 21 ,Interferon-gamma ,Mice ,Th2 Cells ,Antigen ,Dual-specificity phosphatase ,medicine ,Animals ,IL-2 receptor ,Molecular Biology ,Regulation of gene expression ,Mice, Inbred BALB C ,CD40 ,biology ,c-jun ,JNK Mitogen-Activated Protein Kinases ,Acetylation ,Cell Differentiation ,Cell Biology ,Th1 Cells ,Molecular biology ,medicine.anatomical_structure ,Immunoglobulin G ,biology.protein ,Dual-Specificity Phosphatases ,Mitogen-Activated Protein Kinase Phosphatases ,Female ,Interleukin-4 ,T-Box Domain Proteins - Abstract
Naïve CD4(+) T helper (Th) cells differentiate into distinct subsets of effector cells (Th1, Th2, Th17, and induced regulatory T cells (iTreg)) expressing different sets of cytokines upon encounter with presented foreign antigens. It has been well established that Th1/Th2 balance is critical for the nature of the following immune responses. Previous reports have demonstrated important roles of c-Jun N-terminal kinase (JNK) in Th1/Th2 balance, whereas the regulatory mechanisms of JNK activity in Th cells have not been elucidated. Here, we show that dual specificity phosphatase 16 (DUSP16, also referred to as MKP-M or MKP-7), which preferentially inactivates JNK, is selectively expressed in Th2 cells. In the in vitro differentiation assay of naïve CD4(+) cells, DUSP16 expression is up-regulated during Th2 differentiation and down-regulated during Th1 differentiation. Chromatin immunoprecipitation revealed the increased acetylation of histone H3/H4 at the dusp16 gene promoter in CD4(+) T cells under the Th2 condition. Adenoviral transduction of naïve CD4(+) T cells with DUSP16 resulted in increased mRNA expression of IL-4 and GATA-3 in Th2 and decreased expression of IFNγ and T-bet in Th1 differentiation. In contrast, transduction of a dominant negative form of DUSP16 had the reverse effects. Furthermore, upon immunization, T cell-specific dusp16 transgenic mice produced antigen-specific IgG2a at lower amounts, whereas DN dusp16 transgenic mice produced higher amounts of antigen-specific IgG2a accompanied by decreased amounts of antigen-specific IgG1 and IgE than those of control mice. Together, these data suggest the functional role of DUSP16 in Th1/Th2 balance.
- Published
- 2011
42. Molecular mechanisms of the inhibitory effect of lipopolysaccharide (LPS) on osteoblast differentiation
- Author
-
Joji Kusuyama, Aya Maeda, Kenjiro Bandow, Mitsuo Shamoto, Kyoko Kakimoto, Tomokazu Ohnishi, and Tetsuya Matsuguchi
- Subjects
Lipopolysaccharides ,medicine.medical_specialty ,Cellular differentiation ,Biophysics ,Core Binding Factor Alpha 1 Subunit ,Biology ,Biochemistry ,Osteocytes ,Mice ,Osteogenesis ,Internal medicine ,Proto-Oncogene Proteins ,medicine ,Animals ,Sp7 Transcription Factor ,Molecular Biology ,Transcription factor ,Mice, Knockout ,Osteoblasts ,ATF4 ,Osteoblast ,Cell Differentiation ,Cell Biology ,MAP Kinase Kinase Kinases ,Activating Transcription Factor 4 ,Cell biology ,RUNX2 ,Mice, Inbred C57BL ,Toll-Like Receptor 4 ,medicine.anatomical_structure ,Endocrinology ,Myeloid Differentiation Factor 88 ,TLR4 ,Transcription Factors - Abstract
Osteoblasts express Toll like receptor (TLR) 4 and produce osteoclast-activating cytokines in response to the stimulation by lipopolysaccharide (LPS). It has recently been reported that LPS exerts an inhibitory effect on osteoblast differentiation into osteocytes. However, the molecular mechanisms of this inhibitory effect remain ambiguous. The downstream signals of TLR4 are mediated by adaptor molecules including myeloid differentiation factor 88 (MyD88), leading to the activation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinases (ERKs), whose activation by LPS requires the upstream serine/threonine kinase, Cot/Tpl2. To determine the signal molecules responsible for the inhibitory effects of LPS on osteoblast differentiation, we examined the in vitro differentiation of the primary osteoblasts from myd88(-/-) and cot/tpl2(-/-) mice. The matrix mineralization by the wild-type and cot/tpl2(-/-) osteoblasts was significantly inhibited by LPS, whereas that of myd88(-/-) was not affected. During differentiation, LPS suppressed the mRNA expression of runt related transcription factor 2 (Runx2), osterix (Sp7), and activating transcription factor 4 (ATF4) in the wild-type, but not in the myd88(-/-) osteoblasts. The inhibitory effect of LPS on the mRNA expression of these transcription factors was absent in the early phase but partially impaired in the late phase of differentiation in the cot/tpl2(-/-) osteoblasts. Thus, the inhibitory effect of LPS on osteoblast differentiation is Myd88-dependent, whereas the degree of its requirement for Cot/Tpl2 varies depending on the differentiation phase.
- Published
- 2010
43. Osteoblast differentiation is functionally associated with decreased AMP kinase activity
- Author
-
Shinichiro Kawamoto, Tomokazu Ohnishi, Tetsuya Matsuguchi, Kyoko Kakimoto, Norika Chiba, Takayuki Kasai, Eiichi Nagaoka, Kenjiro Bandow, and Hiraku Suzuki
- Subjects
musculoskeletal diseases ,Physiology ,Cell Survival ,Sialoglycoproteins ,Clinical Biochemistry ,Osteocalcin ,Gene Expression ,Core Binding Factor Alpha 1 Subunit ,AMP-Activated Protein Kinases ,Transfection ,Cell Line ,Mice ,Calcification, Physiologic ,stomatognathic system ,3T3-L1 Cells ,medicine ,Adipocytes ,Animals ,Hypoglycemic Agents ,Integrin-Binding Sialoprotein ,Osteopontin ,Phosphorylation ,Protein kinase A ,Cells, Cultured ,Osteoblasts ,biology ,Kinase ,AMPK ,Osteoblast ,Cell Differentiation ,Cell Biology ,Ribonucleotides ,Alkaline Phosphatase ,Aminoimidazole Carboxamide ,Metformin ,Cell biology ,RUNX2 ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Glucose ,Biochemistry ,biology.protein ,Signal transduction - Abstract
Osteoblasts, originating from mesenchymal stem cells, play a pivotal role in bone formation and mineralization. Several transcription factors including runt-related transcription factor 2 (Runx2) have been reported to be essential for osteoblast differentiation, whereas the cytoplasmic signal transduction pathways controlling the differentiation process have not been fully elucidated. AMP-activated protein kinase (AMPK) is a serine-threonine kinase generally regarded as a key regulator of cellular energy homeostasis, polarity, and division. Recent lines of evidence have indicated that the activity of the catalytic alpha subunit of AMPK is regulated through its phosphorylation by upstream AMPK kinases (AMPKKs) including LKB1. Here, we explored the role of AMPK in osteoblast differentiation using in vitro culture models. Phosphorylation of AMPKalpha was significantly decreased during osteoblastic differentiation in both primary osteoblasts and MC3T3-E1, a mouse osteoblastic cell line. Conversely, the terminal differentiation of primary osteoblasts and MC3T3-E1 cells, represented by matrix mineralization, was significantly inhibited by glucose restriction and stimulation with metformin, both of which are known activators of AMPK. Matrix mineralization of MC3T3-E1 cells was also inhibited by the forced expression of a constitutively active form of AMPKalpha. Metformin significantly inhibited gene expression of Runx2 along with osteoblast differentiation markers including osteocalcin (Ocn), bone sialo protein (Bsp), and osteopontin (Opn). Thus, our present data indicate that differentiation of osteoblasts is functionally associated with decreased AMPK activity.
- Published
- 2009
44. JNK activity is essential for Atf4 expression and late-stage osteoblast differentiation
- Author
-
Akio Masuda, Tetsuya Matsuguchi, Kyoko Kakimoto, Tomokazu Ohnishi, Kenjiro Bandow, and Norika Chiba
- Subjects
musculoskeletal diseases ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Cellular differentiation ,Sialoglycoproteins ,Osteocalcin ,Bone Matrix ,Core Binding Factor Alpha 1 Subunit ,Cell Line ,Substrate Specificity ,Mice ,Calcification, Physiologic ,Internal medicine ,medicine ,Animals ,Integrin-Binding Sialoprotein ,Mitogen-Activated Protein Kinase 9 ,Orthopedics and Sports Medicine ,Osteopontin ,Sp7 Transcription Factor ,Transcription factor ,Protein Kinase Inhibitors ,Osteoblasts ,biology ,ATF4 ,Osteoblast ,Cell Differentiation ,Ascorbic acid ,Activating Transcription Factor 4 ,Phosphoric Monoester Hydrolases ,Cell biology ,Clone Cells ,RUNX2 ,Isoenzymes ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Endocrinology ,Gene Expression Regulation ,biology.protein ,Transcription Factors - Abstract
Osteoblasts differentiate from mesodermal progenitors and play a pivotal role in bone formation and mineralization. Several transcription factors including runt-related transcription factor 2 (RUNX2), Osterix (OSX), and activating transcription factor4 (ATF4) are known to be crucial for the process, whereas the upstream signal transduction controlling the osteoblast differentiation sequence is largely unknown. Here, we explored the role of c-jun N-terminal kinase (JNK) in osteoblast differentiation using in vitro differentiation models of primary osteoblasts and MC3T3-E1 cells with ascorbic acid/beta-glycerophosphate treatment. Terminal osteoblast differentiation, represented by matrix mineralization, was significantly inhibited by the inactivation of JNK with its specific inhibitor and exogenous overexpression of MKP-M (MAP kinase phosphatase isolated from macrophages), which preferentially inactivates JNK. Conversely, enhanced mineral deposition was observed by inducible overexpression of p54(JNK2), whereas it was not observed by the overexpression of p46(JNK1) or p46(JNK2), indicating a distinct enhancing role of p54(JNK2) in osteoblast differentiation. Inactivation of JNK significantly inhibited late-stage molecular events of osteoblast differentiation, including gene expression of osteocalcin (Ocn) and bone sialoprotein (Bsp). In contrast, earlier differentiation events including alkaline phosphatase (ALP) activation and osteopontin (Opn) expression were not inhibited by JNK inactivation. Although the expression levels of two transcription factor genes, Runx2 and Osx, were not significantly affected by JNK inactivation, induction of Atf4 mRNA during osteoblast differentiation was significantly inhibited. Taken together, these data indicate that JNK activity is specifically required for the late-stage differentiation events of osteoblasts.
- Published
- 2008
45. Force-induced IL-8 from periodontal ligament cells requires IL-1beta
- Author
-
Aya Maeda, Kazuto Kuroe, Kyoko Kakimoto, Tetsuya Matsuguchi, A. Okamoto, Shouichi Miyawaki, Kenjiro Bandow, and Kazuhisa Soejima
- Subjects
0301 basic medicine ,Dental Stress Analysis ,Chemokine ,Tooth Movement Techniques ,MAP Kinase Signaling System ,Periodontal Ligament ,Interleukin-1beta ,Osteoclasts ,Bone remodeling ,03 medical and health sciences ,0302 clinical medicine ,stomatognathic system ,Shear stress ,Pressure ,Periodontal fiber ,Humans ,Interleukin 8 ,Autocrine signalling ,General Dentistry ,Cells, Cultured ,biology ,Chemistry ,Interleukin-8 ,030206 dentistry ,Anatomy ,Gingival Crevicular Fluid ,In vitro ,Cell biology ,030104 developmental biology ,Mitogen-activated protein kinase ,biology.protein ,Bone Remodeling ,Stress, Mechanical ,Shear Strength - Abstract
During orthodontic tooth movement, mechanical stresses induce inflammatory reactions in the periodontal ligament (PDL). We hypothesized that chemokines released from PDL cells under mechanical stress regulate osteoclastogenesis, and investigated the profiles and mechanisms of chemokine expression by human PDL cells in response to mechanical stress. In vitro, shear stress and pressure force rapidly increased the gene and protein expressions of IL-8/CXCL8 by PDL cells. Consistently, amounts of IL-8 in the gingival crevicular fluid of healthy individuals increased within 2 to 4 days of orthodontic force application. The PDL cells constitutively expressed low levels of IL-1β, which were not further increased by mechanical stress. Interestingly, neutralization of IL-1β abolished IL-8 induction by mechanical stresses, indicating that IL-1β is essential for IL-8 induction, presumably though autocrine or paracrine mechanisms. Finally, experiments with signal-specific inhibitors indicated that MAP kinase activation is essential for IL-8 induction.
- Published
- 2007
46. I–2 Low Intensity Pulsed Ultrasound (LIPUS) Helps to Maintain the Undifferentiated Status of Mesenchymal Stem Cells
- Author
-
Tetsuya Matsuguchi, Kenjiro Bandow, Joji Kusuyama, Tomokazu Ohnishi, Kyoko Kakimoto, and Chang Hwan Seong
- Subjects
business.industry ,Mesenchymal stem cell ,Medicine ,Orthopedics and Sports Medicine ,Surgery ,General Medicine ,Low-intensity pulsed ultrasound ,business ,Regenerative medicine ,Cell biology - Abstract
Mesenchymal Stem Cells (MSCs) are pluripotent cells with the capability of self-renewal. It has been established that MSCs can transform into osteoblasts, adipocytes or chondrocytes depending on the culture condition. Regenerative medicine is considered to become an essential therapy in the field of
- Published
- 2015
47. Low-intensity pulsed ultrasound (LIPUS) induces RANKL, MCP-1, and MIP-1beta expression in osteoblasts through the angiotensin II type 1 receptor
- Author
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Tetsuya Matsuguchi, Sadahiro Iwabuchi, Kazuto Kuroe, Kyoko Kakimoto, Kenjiro Bandow, Yoshiaki Nishikawa, Tomokazu Ohnishi, and Kazuhisa Soejima
- Subjects
MAPK/ERK pathway ,medicine.medical_specialty ,Time Factors ,Physiology ,Ultrasonic Therapy ,Clinical Biochemistry ,Tetrazoles ,Bone healing ,Low-intensity pulsed ultrasound ,Mechanotransduction, Cellular ,Receptor, Angiotensin, Type 1 ,Bone remodeling ,Mice ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Phosphorylation ,Receptor ,Chemokine CCL4 ,Extracellular Signal-Regulated MAP Kinases ,Chemokine CCL5 ,Chemokine CCL2 ,Angiotensin II receptor type 1 ,Osteoblasts ,biology ,Chemistry ,Biphenyl Compounds ,RANK Ligand ,Osteoprotegerin ,Cell Differentiation ,Cell Biology ,Angiotensin II ,Cell biology ,Up-Regulation ,Endocrinology ,RANKL ,Chemokines, CC ,biology.protein ,Benzimidazoles ,Stress, Mechanical ,Angiotensin II Type 1 Receptor Blockers - Abstract
Constant mechanical stress is essential for the maintenance of bone mass and strength, which is achieved through the cooperative functions of osteoblasts and osteoclasts. However, it has not been fully elucidated how these cell types mediate mechanical signals. Low-intensity pulsed ultrasound (LIPUS) therapy is a recently developed method for application of mechanical stress, and is used clinically to promote bone fracture healing. In the present study, we applied LIPUS to osteoblasts at different stages of maturation and analyzed their chemokine and cytokine expression. In comparison with their immature counterparts, mature osteoblasts expressed significantly higher levels of mRNAs for the receptor activator of nuclear factor kappa B ligand (RANKL), monocyte chemoattractant protein (MCP)-1, and macrophage-inflammatory protein (MIP)-1beta after a few hours of LIPUS treatment. Intriguingly, protein and mRNA expression of angiotensin II type 1 receptor (AT1), a known mechanoreceptor in cardiomyocytes, was detected in osteoblasts, and the level of expression increased significantly during cell maturation. Furthermore, LIPUS-induced extracellular signal-regulated kinase (ERK) phosphorylation and RANKL/chemokine expression was abrogated by a specific AT1 inhibitor. Thus, AT1 may play one of the essential roles in bone metabolism as a mechanoreceptor of osteoblasts.
- Published
- 2006
48. Mature hepatocyte growth factor/scatter factor on the surface of human granulocytes is released by a mechanism involving activated factor Xa
- Author
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Kyoko Kakimoto, Kenjiro Bandow, Charles J. Lowenstein, Tomokazu Ohnishi, Yasushi Daikuhara, and Tetsuya Matsuguchi
- Subjects
Serum ,Cell type ,Immunology ,Cell ,Granulocyte ,Biology ,Nitric Oxide ,Cell Degranulation ,Cell Line ,Thromboplastin ,In vivo ,medicine ,Immunology and Allergy ,Animals ,Humans ,Blood Coagulation ,Wound Healing ,Cell-Free System ,Hepatocyte Growth Factor ,Cell Membrane ,Serum Albumin, Bovine ,Molecular biology ,In vitro ,medicine.anatomical_structure ,Coagulation ,Factor Xa ,Hepatocyte growth factor ,Cattle ,medicine.drug ,Granulocytes ,Protein Binding - Abstract
Serum hepatocyte growth factor (HGF) is rapidly increased in patients suffering from various tissue injuries including arterial occlusive diseases. However, the cellular sources of the HGF increase remain largely unknown. In the present study, we showed that bioactive mature HGF is constitutively present on the surface of granulocytes in human peripheral blood. Exogenously added 125I-labeled iodo-HGF efficiently bound to granulocyte surface, whereas only a scarce amount of HGF mRNA was detected in granulocytes, indicating that the mature HGF on granulocytes is likely to be derived from other cell types. Interestingly, treatment of granulocytes with human serum rapidly induced the release of the cell surface-associated HGF. In vivo, thromboplastin injection into mice increased HGF release from transplanted human granulocytes, which was inhibited by the pretreatment with DX9065a, a specific inhibitor of factor Xa. Furthermore, DX9065a also inhibited the serum-induced HGF release from human granulocytes in vitro, suggesting that the HGF-releasing factor(s) in serum is associated with factor Xa activation. Thus, human granulocytes may function as a transporter of HGF in the peripheral blood, releasing HGF at the injured sites caused by blood coagulation, where HGF may promote tissue repair.
- Published
- 2006
49. I–2 Osteoblast Responses to Low-Intensity Pulsed Ultrasound (LIPUS) are Attenuated by Osteopontin Expression
- Author
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Joji Kusuyama, Kenjiro Bandow, Tetsuya Matsuguchi, Tomokazu Ohnishi, Kyoko Kakimoto, and Mitsuo Shamoto
- Subjects
medicine.anatomical_structure ,biology ,business.industry ,biology.protein ,Medicine ,Orthopedics and Sports Medicine ,Surgery ,Osteoblast ,General Medicine ,Osteopontin ,Low-intensity pulsed ultrasound ,business ,Cell biology - Published
- 2014
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