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Cigarette smoke (CS) exposure is a risk factor for chronic obstructive pulmonary disease (COPD). CS exposure impairs the ability of killing pathogens in macrophages, which might be due to the abnormal apoptosis induced by CS. This study explored the effects and mechanisms of cigarette smoke extract (CSE) on the apoptosis of macrophages in vitro. Raw264.7 cells were treated with CSE at different concentrations, and viability and apoptosis of cells was accessed. The protein expression was detected by western blot. The intracellular Ca 2+ level was evaluated by Fluo-4 AM probe assay. CSE induced the apoptosis and increased the expression of cleaved caspase 3, which were attenuated by a caspase inhibitor. CSE increased the expression of CHOP, BiP and P-eif2α, and the inhibitor of endoplasmic reticulum stress (ERS) decreased the apoptosis induced by CSE. Phosphorylation levels of P38, JNK and ERK1/2 were increased following incubation with CSE. Only P38 inhibitor significantly reduced apoptosis induced by CSE, while ERK1/2 inhibitor promoted apoptosis. Phosphorylation of STAT1 at Ser727 was activated by CSE and attenuated by the P38 inhibitor. Finally, CSE increased the level of intracellular Ca 2+ , and calcium chelator partly attenuated the apoptosis and phosphorylation of P38 and STAT1 induced by CSE. CSE induced a caspase 3-dependent apoptosis in Raw264.7 cells via ERS and intracellular Ca 2+ /P38/STAT1 pathway., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Type 2 diabetes, a global health concern has been considered as major risk factor for cardiovascular diseases. Hinokinin, an emerging bioactive lignin, is reported to show wide range of pharmacological activities. However, the protective role and mechanisms of Hinokinin against type 2 diabetes-mediated cardiotoxicity are still remains unknown. An experimental type 2 diabetic mice model was created by treating animals with high fat diet for four weeks and intraperitoneal injection of streptozotocin (35 mg/kg body weight). Post-type 2 diabetic induction, animals orally treated with Hinokinin (20 or 40 mg/kg body weight) for six weeks. The type 2 diabetic mice exhibited a rise in blood glucose level as well as glycated hemoglobin (HbA1c %), decrease in weekly body weights, decrease in food intake, reduction in absolute heart weight, fall in serum insulin level with altered lipid profile and cardiac functional damage. Diabetic mice treated with Hinokinin attenuated hyperglycemia, dyslipidemia and cardiac dysfunction. In addition, Hinokinin ameliorated histological alterations, fibrosis and glycated proteins in HFD/STZ-induced mice. Type 2 diabetic condition in mice exacerbated oxidative stress, inflammatory status and apoptosis. Hinokinin treatment significantly assuaged oxidative stress, inflammation and apoptosis and elevated antioxidant defenses in diabetic heart. The underlying mechanisms for such mitigation involved the modulation of Nrf2/Keap1/ARE pathway, MAPKs (JNK, p38 and ERK 1/2) and TLR4/MyD88/NF-κB mediated inflammatory pathways and mitochondrial-dependent (intrinsic) apoptosis pathway. In conclusion, the results of this study provided clear evidence that Hinokinin protects against HFD/STZ (type 2 diabetes)-induced cardiac injury by alleviating oxidative stress, inflammation and apoptosis., (Copyright © 2021. Published by Elsevier Masson SAS.)

Background: The active components of Dracocephalum moldavica L. (TFDM) can inhibit myocardial ischemia by inhibiting oxidative stress. However, the effects of TFDM on astrocytes have not been investigated in vitro. The current study aimed to explore whether TFDM protects astrocytes against H 2 O 2 -induced apoptosis through a mitochondria-dependent pathway., Methods: The human glioma cell line U87 was used to investigate the ability of TFDM to protect astrocytes against H 2 O 2 -induced apoptosis. The cell counting kit-8 assay and flow cytometry were used to detect cell viability, apoptosis, MMP, Ca 2+ influx and reactive oxygen species (ROS). Lactate dehydrogenase (LDH) and malonic dialdehyde (MDA) levels were measured by ELISA. In addition, protein and mRNA expression changes were detected by Western blotting and qRT-PCR., Results: TFDM (0.78~200 μg/ml) had limited cytotoxic effects on the viability of U87 cells. Compared with the model group (treated with H2O2 only), cells treated with medium- and high-dose TFDM exhibited reduced MDA concentrations (P < 0.05) and ROS production (P < 0.05) and decreased MMP (P < 0.05) and reduced apoptosis (P < 0.05). The percentage of annexin V-FITC-stained cells was markedly suppressed by TFDM, confirming its anti-apoptotic properties. WB results showed that protein expression of Bcl-2-associated X protein (BAX), Caspase-3, Caspase-9, Caspase-12, and B-cell leukemia/lymphoma 2 (Bcl2) was reduced in the TFDM group compared with that in the model group (P < 0.05) and that expression of these proteins was normalized by TFDM treatment in a dose-dependent manner. According to RT-qPCR results, TFDM pretreatment resulted in reduced mRNA expression of BAX, Caspase-9, Caspase-12, p38MAPK, and CaMKII and increased mRNA expression of mTOR compared with the model group., Conclusions: The current study revealed the protective effects of TFDM on U87 cells under oxidative stress conditions through the inhibition of a mitochondria-dependent pathway that is associated with the CaMKII/P38MAPK/ERK1/2 and PI3K/AKT/mTOR pathways.

Lipopolysaccharide (LPS) can regulate the expression of apoptotic factors, including caspase‑3, Bcl‑2 and Bcl‑2‑associated X protein (Bax). Nitric oxide (NO) plays an important role in apoptosis. N‑acetyl cysteine (NAC) has been shown to exhibit antioxidant effects in vitro. However, the effects of NAC on LPS‑induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the associated mechanisms are not well characterized. The present study explored the effect of NAC on LPS‑induced apoptosis of HUVECs and determined the participation of the p38 mitogen‑activated protein kinase (MAPK) pathway in the process of apoptosis. Cell viability was assessed using the Cell Counting Kit‑8 (CCK‑8) assay. The expression of caspase‑3, Bax, Bcl‑2, phosphorylated (p)‑p38MAPK/total (t‑)p38MAPK and p‑endothelial e nitric oxide synthase (eNOS)/t‑eNOS proteins were determined by western blotting. The expression levels of caspase‑3, Bax and Bcl‑2 mRNA were determined using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). The rate of apoptosis was determined using flow cytometry. An NO detection kit (nitric reductase method) was used to determine NO concentration. The results of CCK‑8 and flow cytometric analyses showed that pretreatment of HUVECs with NAC or p38MAPK inhibitor (SB203580) attenuated LPS‑induced decrease in cell viability and increase in cell apoptosis. RT‑qPCR and western blotting showed that LPS promoted caspase‑3 and Bax expression, but inhibited that of Bcl‑2 in HUVECs; however, these effects were attenuated by pretreatment with NAC or SB203580. LPS stimulation significantly enhanced the expression of p‑p38MAPK protein and reduced the expression of p‑eNOS protein; however, these effects were attenuated by pretreatment with NAC or SB203580. NAC pretreatment attenuated LPS‑induced inhibition of NO synthesis, which was consistent with the effects of SB203580. The results demonstrated that NAC pretreatment alleviated LPS‑induced apoptosis and inhibition of NO production in HUVECs. Furthermore, these effects were proposed to be mediated via the p38MAPK signaling pathway.

Background: To study the effect of parecoxib, a novel cyclooxygenase-2 selective inhibitor, on the radiation response of colorectal cancer (CRC) cells and its underlying mechanisms., Materials and Methods: Both in vitro colony formation and apoptosis assays as well as in vivo mouse xenograft experiments were used to explore the radiosensitizing effects of parecoxib in human HCT116 and HT29 CRC cells., Results: Parecoxib sensitized CRC cells to radiation in vitro with a sensitivity enhancement ratio of 1.32 for HCT116 cells and 1.15 for HT29 cells at a surviving fraction of 0.37. This effect was partially attributable to enhanced apoptosis induction by parecoxib combined with radiation, as illustrated using an in vitro apoptosis assays. Parecoxib augmented the tumor response of HCT116 xenografts to radiation, achieving growth delay more than 20 days and an enhancement factor of 1.53. In accordance with the in vitro results, parecoxib combined with radiation resulted in less proliferation and more apoptosis in tumors than radiation alone. Radiation monotherapy decreased microvessel density (MVD) and microvessel intensity (MVI), but increased the hypoxia level in xenografts. Parecoxib did not affect MVD, but it increased MVI and attenuated hypoxia., Conclusions: Parecoxib can effectively enhance radiation sensitivity in CRC cells through direct effects on tumor cells and indirect effects on tumor vasculature.

Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer-related mortality in females worldwide. The efficacy of chemotherapeutic drugs on breast cancer is hindered by many factors, including that cancer stem-like side population (SP) cells may contribute to tumorigenesis and resistance to chemotherapy. Doxorubicin and gemcitabine are two of the most effective and widely used chemotherapeutic agents for the treatment of various human malignancies. To the best of our knowledge, the effects of doxorubicin and gemcitabine on breast cancer SP cells are poorly understood. In the present study, we examined the potential roles of doxorubicin and gemcitabine in breast cancer main population (MP) and SP cells, and the mechanisms of doxorubicin and gemcitabine. The results showed that doxorubicin and gemcitabine decreasede proliferation, and increased apoptosis in the MCF7 MP and SP cells in vitro. Furthermore, doxorubicin and gemcitabine inhibited tumor growth and improved the survival rate of mice in vivo. Additionally, the mechanisms of doxorubicin and gemcitabine in MCF7 MP and SP cells were determined.

Objective: To investigate the effects of anti-CD44 mAb A3D8 on the cell proliferation of human acute monocytic leukemia cell line THP-1 and its mechanism., Methods: Cell proliferation was assayed with MTT method, the expression of CD33, CD15, CD11b, CD14, Annexin-V, caspase-3 and cell cycle with flow cytometry, and the expression of p-Akt, p-ERK, bcl-2 and p27kip1 with Western blot., Results: A3D8 could remarkably inhibit the proliferation capacity of the THP-1 cells in a dosage- and time-dependent manner. THP-1 differentiation was observed when treated with A3D8 (2.0 µg/ml) for one to six days. Expression of CD33 (68.9 ± 2.0 vs 39.3 ± 1.5), CD15 (61.7 ± 5.5 vs 12.9 ± 2.6), CD11b (67.3 ± 3.8 vs 14.0 ± 2.0) and CD14 (83.0 ± 5.7 vs 8.0 ± 1.0) was significantly increased at day 4 compared with the control group (all P < 0.01). Cell cycle of the THP-1 cells was arrested in G(0)/G(1). Expression of the Annexin-V \[(32.5 ± 2.5)% vs (2.4 ± 0.3)%\] and caspase-3 \[(33.3 ± 2.5)% vs (3.6 ± 0.3)%\] was much higher than that in normal controls (all P < 0.01), and apoptosis was observed in THP-1 cells at day 5. Expression of p-Akt (0.24 ± 0.06 vs 1.20 ± 0.15), p-ERK (0.32 ± 0.05 vs 1.24 ± 0.09), and bcl-2 (0.11 ± 0.05 vs 0.65 ± 0.07) was much lower than that of the controls (all P < 0.01), while p27kip1 (1.08 ± 0.09 vs 0.10 ± 0.02) was significantly increased at day 4 (P < 0.05)., Conclusion: Anti-CD44 antibody can induce the differentiation and apoptosis of THP-1 cell through inhibiting PI3K/AKt and ERK1/2 signaling pathway.

Oxidative stress is one of the several mechanisms that induces apoptosis in cells. It has been shown that hydrogen peroxide (H(2)O(2)) induces apoptosis in several kinds of cells; however, the role of H(2)O(2) in the apoptosis of Schwann cells (SCs) is currently unclear. The objective of this study was to determine whether H(2)O(2) is capable of inducing apoptosis in SCs and whether or not such an effect is associated with the activation of mitochondrial pathway. We demonstrated that H(2)O(2) induces apoptosis in SCs, and is associated with increased release of cytochrome c from mitochondria and the activation of caspase-3 and -9 by up-regulation of Bax and down-regulation of Bcl-2. These results suggest a potential role for H(2)O(2) in SC injury by triggering apoptosis via the mitochondrial pathway under oxidative stress., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

The aim of this study was to investigate the effect of rapamycin on cell growth and apoptosis in the myelodysplastic syndrome (MDS) cell line MUTZ-1 and possible mechanism. MUTZ-1 cells were treated with rapamycin, cell proliferation capability was determined with MTT, protein expression including Annexin V/PI, caspase 3, PTEN, p-Akt, p-mTOR and the cell cycle were analyzed with flow cytometry. The results indicated that the proliferation of MUTZ-1 cells was inhibited by rapamycin in concentration-and time-dependent manners (r=0.67, 0.61, 0.72). After treatment with rapamycin for 24-72 hours, cell count in G0/G1 were significantly higher than that of the control (p<0.01), and this effect showed a time-and concentration-dependency (r=0.94, 0.93, 0.92), the cell cycle was blocked in G0/G1 phase. As compared with control group, the proportion of Annexin V+PI-MUTZ-1 cells and the cellular PTEN levels increased in the treated group dramatically and in time-and dose-dependent manners (p<0.01). To the contrary, level of p-mTOR expression markedly decreased as compared with control group (p<0.05). It is concluded that the rapamycin inhibits the proliferation of MUTZ-1 cells, down-regulates the PTEN/PI3K-Akt/mTOR signaling pathway by interaction with mTOR, which induces the apoptosis of mUTZ-1 cells.

Aim of the study: Cardiovascular disease (CVD) seriously endangers human health and is characterized by high mortality and disability. The effectiveness of Dracocephalum moldavica L. in the treatment of CVD has been proven by clinical practice. However, the mechanism by which DML can treat CVD has not been systematically determined. Materials and methods: The active compounds in DML were screened by literature mining and pharmacokinetic analysis. Cytoscape software was used to construct the target-disease interaction network of DML in the treatment of CVD. Gene ontology and signalling pathway enrichment analyses were performed. The key target pathway network of DML compounds was constructed and verified by pharmacological experiments in vitro. A hydrogen glucose deprivation/reoxygenation model was established in H9c2 cells using hypoxia and glucose deprivation for 9 h combined with reoxygenation for 2 h. The model simulated myocardial ischaemic reperfusion injury to investigate the effects of total flavonoids of Cymbidium on cell viability, myocardial injury markers, oxidative stress levels, and reactive oxygen radical levels. Western blot analysis was used to examine NOX-4, Bcl-2/Bax, and PGC-1α protein expression. Results: Twenty-seven active components were screened, and 59 potential drug targets for the treatment of CVD were obtained. Through the compound-target interaction network and the target-disease interaction network, the key targets and key signalling pathways, such as NOX-4, Bcl-2/Bax and PGC-1α, were obtained. TFDM significantly decreased LDH and MDA levels and the production of ROS and increased SOD activity levels in the context of OGD/R injury. Further studies indicated that NOX-4 and Bax protein levels and the p-P38 MAPK/P38 MAPK andp-Erk1/2/Erk1/2 ratios were suppressed by TFDM. The protein expression of Bcl-2 and PGC-1α was increased by TFDM. Conclusions: Our results showed that DML had multicomponent, multitarget and multichannel characteristics in the treatment of CVD. The mechanism may be associated with the following signalling pathways: 1) the NOX-4/ROS/p38 MAPK signalling pathway, which inhibits inflammation and reactive oxygen species (ROS) production, and 2) the Bcl-2/Bax and AMPK/SIRT1/PGC-1α signalling pathways, which inhibit apoptosis. [ABSTRACT FROM AUTHOR]

BACKGROUND: Sacha inchi is known for its high protein content and medicinal properties. Bioactive peptides have been reported to have therapeutic potential in various human diseases. However, there is a lack of research evaluating the pharmacological value of peptides derived from Sacha inchi. Therefore, this study aimed to investigate the anti‐hepatoma effect of Sacha inchi peptides (SPs) and their underlying mechanism. RESULTS: The study found that treatment with SPs significantly reduced the proliferation of HepG2 cells by inducing apoptosis and arresting the cell cycle at the G0/G1 phase. SPs also induced HepG2 cell apoptosis by increasing the levels of proteins such as Bax, Caspase‐3 and P53. The study identified nine novel peptides in SPs, of which LLEPDVR, ALVEKAKAS and TGDGSLRPY exhibited higher cell proliferative inhibition rates compared to other peptides. CONCLUSION: The findings of this study suggest that Sacha inchi peptides have potential pharmacological effects in the treatment of liver cancer. SPs effectively suppress the cell cycle and facilitate cell apoptosis, indicating their anti‐hepatoma effect. The novel peptides identified in SPs may have therapeutic value for liver cancer treatment. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Simple Summary: ASCT2 is an attractive tumour metabolism target based on its critical role in cancer cell growth. The potential mechanisms of microtubule protein inhibitor C118P in breast cancer remain unknown. Identification of the potential target of C118P is essential. We evaluated the inhibitory effects of C118P on breast cancer. C118P restrained the tumour growth of MDA-MB-231 cells by inducing apoptosis, G2/M phase arrest, and autophagy. Furthermore, ASCT2 was confirmed to be a target of C118P. This is the first report to provide evidence that ASCT2 might be a candidate target of C118P in breast cancer treatment. Remarkably, this study will provide an opportunity for ASCT2 inhibition as a therapeutic strategy. Background: The microtubule protein inhibitor C118P shows excellent anti-breast cancer effects. However, the potential targets and mechanisms of C118P in breast cancer remain unknown. Methods: Real-time cellular analysis (RTCA) was used to detect cell viability. Apoptosis and the cell cycle were detected by flow cytometry. Computer docking simulations, surface plasmon resonance (SPR) technology, and microscale thermophoresis (MST) were conducted to study the interaction between C118P and alanine-serine-cysteine transporter 2 (ASCT2). Seahorse XF technology was used to measure the basal oxygen consumption rate (OCR). The effect of C118P in the adipose microenvironment was explored using a co-culture model of adipocytes and breast cancer cells and mouse cytokine chip. Results: C118P inhibited proliferation, potentiated apoptosis, and induced G2/M cell cycle arrest in breast cancer cells. Notably, ASCT2 was validated as a C118P target through reverse docking, SPR, and MST. C118P suppressed glutamine metabolism and mediated autophagy via ASCT2. Similar results were obtained in the adipocyte–breast cancer microenvironment. Adipose-derived interleukin-6 (IL-6) promoted the proliferation of breast cancer cells by enhancing glutamine metabolism via ASCT2. C118P inhibited the upregulation of ASCT2 by inhibiting the effect of IL-6 in co-cultures. Conclusion: C118P exerts an antitumour effect against breast cancer via the glutamine transporter ASCT2. [ABSTRACT FROM AUTHOR]

Rhizopus arrhizus is a fungus that can cause central nervous system infections in animals, resulting in high morbidity and mortality, but the mechanism of injury is rarely reported. In this study, we investigated the mechanism of Rhizopus arrhizus damage to the central nervous system of mice by observing the clinical neurological symptoms and resolving the pathological changes in the ultrastructure of brain tissues, combined with the alteration of apoptosis-related genes and immunohistochemistry (IHC). The results showed that all the mice in the treated group died, the brain pyknosis of neurons, there were black mycelium aggregates around the blood vessels, and apoptotic vesicles were produced. The RT-qPCR results showed that, compared with the control group, the relative transcriptome levels of Caspase 8 and BcL-2 genes were significantly increased (P < 0.05), the relative transcriptome level of Caspase 9 gene was highly significant (P < 0.01), the relative transcriptome level of Caspase 3 and Bax gene was significantly decreased (P < 0.05), and the ratio of Bcl-2/Bax was significantly increased (P < 0.05) in the brains of the treated group. TUNEL staining showed that the rate of neuronal apoptosis in the treated group of mice was extremely significantly higher than that in the control group (P < 0.01). This study shows that Rhizopus arrhizus strain XMLO1 causes brain damage by triggering neuronal apoptosis. This study provided a theoretical basis for revealing the mechanism of Rhizopus arrhizus infection. • After infection with Rhizopus arrhizus brain neurons produce apoptotic bodies. • R. arrhizus infection can lead to an increase in brain basophils. • Infection with R. arrhizus can cause changes in apoptotic genes and proteins. [ABSTRACT FROM AUTHOR]

Context: Genistein, one of the isoflavones derived from soybean seeds, has been reported to exert multiple bioactivities. However, the mechanism of its action on the central nervous system is not fully understood. Objective: To investigate the cytoprotection of genistein and its molecular mechanism against H2O2-induced cell death in primary rat cortical neurons. Materials and methods: Genistein (0.01, 0.1, and 1 μM) were added into the primary rat neurons 24 h before and co-cultured with 500 μM H2O2 for 1 h. Neuronal injury was assessed by MTT, lactate dehydrogenase (LDH) assay, and Hoechst33258 staining. Intracellular reactive oxygen species (ROS) generation induced by H2O2 was determined. Neuronal apoptosis was evaluated by Bcl-2/Bax ratio as well as by caspase-9 and caspase-3 activities. The protein levels and phosphorylation of NF-κB/p65, IκB, JNK, and ERK were detected by western blots. Results: Genistein pretreatment attenuated H2O2-mediated neuronal viability loss, nuclear condensation, and ROS generation in a concentration-dependent manner. Genistein exerted anti-apoptotic effects by reversing the apoptotic factors Bcl-2 and Bax ratio, along with the suppression of caspase-9 and caspase-3 activities. In addition, genistein down-regulated the expression of NF-κB/p65, and suppressed the phosphorylation of p65 and IκB. Genistein also inhibited H2O2-induced activation of the MAPK-signaling pathway including JNK and ERK. Discussion and conclusion: The results indicated that genistein effectively protects cortical neurons against oxidative stress at least partly via inactivation of NF-κB as well as MAPK-signaling pathways, and suggested the possibility of this antioxidant for the prevention and treatment of stroke. [ABSTRACT FROM AUTHOR]

PURPOSE: Propranolol was found by chance very effective in treating hemangioma, yet the therapeutic mechanism remain unclear. This study was aimed to investigate the therapeutic mechanism of propranolol on hemangioma. METHODS: The plasma propranolol concentrations at 2,6,10,24 h were assayed by reversed phase high performance liquid chromatography(RP-HPLC)after oral administration of propranolol(l mg/kg*d) once a day(qd, n=6) or twice a day (bid, n=6); The hemangioma-derived stem cells(HemSCs) were isolated from Chinese IH patients. The effect of propranolol on proliferation, apoptosis rate of HemSCs was observed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and bi -fluorescence flow cytometry (FCM).With RT-PCR, real-time PCR, Elisa and Western blot, expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) was observed after treatment with different concentrations (0, 0.02, 0.2, 2.20 μM) of PRN at different times. For in vivo study, HemSCs were treated with PRN, and then subcutaneously injected into BALB/c -nu mice with human umbilical vein endothelial cell (HUVEC). The grafts were harvested after 1 week for HE staining and immunohistological staining. SPSS 17.0 software package was used for statistical analysis. RESULTS: The peak propranolol concentration reached at 2h in both qd group (207.8 ±33.9 ng/mL) and bid group (155.2 ±40.6 ng/mL),the elimination half-life (t1/2) was longer than 6h.The mean concentration of lOh was significantly decreased in qd group, while that was elevated in bid group. The proliferation and total apoptosis rate of HemSCs was not obviously affected by PRN in normal concentration; but the expression of VEGF and bFGF in HemSC was suppressed by PRN. In vivo model,the mean microvessel density (MVD) was significantly decreased after HemSCs were treated by PRN. CONCLUSION: PRN can inhibit HemSC to express VEGF and bFGF, but can't inhibit proliferation or induce apoptosis of HemSCs. Supported by National Natural Science Foundation of China (81070846) and Youth Science Fund of Guangxi Zhuang Autonomous Region (0832044). [ABSTRACT FROM AUTHOR]