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Tumor necrosis factor alpha (TNF-α)/hypoxia-treated 3T3-L1 adipocytes have been used to model inflamed and insulin-resistant adipose tissue: this study examines gaps in the model. We tested whether modulating TNF-α/hypoxia treatment time could reduce cell death while still inducing inflammation and insulin resistance. Adipocytes were treated with TNF-α (12 h or 24 h) and incubated in a hypoxic chamber for 24 h. To examine maintenance of the phenotype over time, glucose and FBS were added at 24 h post initiation of treatment, and the cells were maintained for an additional 48 h. Untreated adipocytes were used as a control. Viability, insulin resistance, and inflammation were assessed using Live/Dead staining, RT-qPCR, ELISA, and glucose uptake assays. Treatment for 12 h with TNF-α in the presence of hypoxia resulted in an increase in the percentage of live cells compared to 24 h treated cells. Importantly, insulin resistance and inflammation were still induced in the 12 h treated adipocytes: the expression of the insulin sensitive and inflammatory genes was decreased and increased, respectively. In 72 h treated adipocytes, no significant differences were found in the viability, glucose uptake or insulin-sensitive and inflammatory gene expression. This study provides a modified approach to in vitro odeling adipocyte inflammation and insulin resistance. [ABSTRACT FROM AUTHOR]

Copyright of Shandong Medical Journal is the property of Shandong Medical Health Newspapers and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Flavanones, a class of flavonoids, are abundant in fruits, vegetables, and herbs. They are known to have several biological activities, such as anti-inflammatory and anti-cancer activities, but their effects on obesity remain unclear. Obesity is closely associated with adipocyte differentiation and lipid accumulation in adipose tissue. Therefore, in this study, we examined the effects of flavanone derivatives on adipocyte differentiation and lipid accumulation by using 3T3-L1 cells. Among the 15 flavanone derivatives studied, 4′-phenylflavanone (4PF), with a biphenyl structure, significantly inhibited adipocyte differentiation-related lipid accumulation in 3T3-L1 cells; this inhibition of lipid accumulation was dose-dependent. Gene expression analysis showed that 4PF suppressed the expression of adipogenic marker genes. Although the induction of peroxisome proliferator activator γ2 (Pparγ2), a master regulator of adipocyte differentiation, and its target genes during adipocyte differentiation was attenuated in 4PF-treated cells, 4PF did not directly regulate Pparγ2 gene expression and its activation. In contrast, 4PF suppressed mitotic clonal expansion (MCE), which is associated with changes in the expression of proliferation-related genes at the early stages of adipocyte differentiation. Taken together, these results suggest that 4PF inhibits lipid accumulation because it suppresses MCE during adipocyte differentiation. Thus, our findings may help in the development of anti-obesity drugs. [ABSTRACT FROM AUTHOR]

Background: The leaves of the olive plant (Olea europaea) contain significantly higher levels of phenolic compounds than the fruit and olive oil, acting as a natural supplement for antioxidant activities. Objectives: This research aimed to investigate the effect of this extract on certain inflammatory factors (TNF-α, IL-1, IL-6) and its regulatory role on tissue gene (RBP4) in fibroblast cell lines (3T3-L1). Methods: After culturing adipose tissue fibroblast cells (3T3-L1) under sterile and standard conditions, the cells were exposed to different logarithmic concentrations of the extract. The production of inflammatory cytokines TNF-α, IL-1, and IL-6 was measured using ELISA methods, and the expression level of the RBP4 gene was assessed using a real-time PCR technique. Results: The results showed dose-dependent effects of the extract on the 3T3-L1 cells. The MTT assay indicated that the hydroalcoholic extract of olive plant leaves has a concentration-dependent effect on cell proliferation (∗∗P ≤ 0.05). The DPPH test results showed a significant decrease in the level of active oxygen in the presence of Olive leaf extract (OLE). The catalase and MDA test results also indicated an increase in antioxidant activity in cells treated with the extract. Conversely, the expression levels of inflammatory cytokines IL-1 and IL-6 increased by 0.05 and 0.01 (pg/mL), respectively, while changes in TNF-α and RBP4 gene expression were insignificant (P > 0.05). Conclusions: The results demonstrated that OLE can enhance the efficiency of antioxidant and inflammatory pathways in the cell and significantly reduce the level of active oxygen in the cell. [ABSTRACT FROM AUTHOR]

ATAD3 is a vital ATPase of the inner mitochondrial membrane of pluri‐cellular eukaryotes, with largely unknown functions but early required for organism development as necessary for mitochondrial biogenesis. ATAD3 knock‐down in C. elegans inhibits at first the development of adipocyte‐like intestinal tissue so we used mouse adipocyte model 3T3‐L1 cells to analyze ATAD3 functions during adipogenesis and lipogenesis in a mammalian model. ATAD3 function was studied by stable and transient modulation of ATAD3 expression in adipogenesis‐ induced 3T3‐L1 cells using Knock‐Down and overexpression strategies, exploring different steps of adipocyte differentiation and lipogenesis. We show that (i) an increase in ATAD3 is preceding differentiation‐induced mitochondrial biogenesis; (ii) downregulation of ATAD3 inhibits adipogenesis, lipogenesis, and impedes overexpression of many mitochondrial proteins; (iii) ATAD3 re‐expression rescues the phenotype of ATAD3 KD, and (iv) differentiation and lipogenesis are accelerated by ATAD3 overexpression, but inhibited by expression of a dominant‐negative mutant. We further show that the ATAD3 KD phenotype is not due to altered insulin signal but involves a limitation of mitochondrial biogenesis linked to Drp1. These results demonstrate that ATAD3 is limiting for in vitro mitochondrial biogenesis and adipogenesis/lipogenesis and therefore that ATAD3 mutation/over‐ or under‐expression could be involved in adipogenic and lipogenic pathologies. [ABSTRACT FROM AUTHOR]

The global prevalence of obesity is rising year by year, which has become a public health problem worldwide. Many animal and clinical studies have shown that Lactiplantibacillus plantarum is considered an ideal probiotic and potential supplement for the treatment of obesity. In this study, we aimed to complete the genome sequence of L. plantarum HOM2217, which was isolated from human milk, and study its physiological characteristics and anti-obesity effects in 3T3-L1 cells and rats fed a high-fat diet (HFD) to determine its potential as a starter for functional food products. Whole-genome analysis demonstrated that HOM2217 contained a single circular chromosome of 3,267,529 bp with a GC content of 44.5% and one plasmid (62,350 bp) with a GC content of 38.5%. Compared to the reference strains, HOM2217 demonstrated superior tolerance to gastrointestinal conditions, higher adhesion to intestinal epithelial cell lines, potent antimicrobial activity against Enterobacter cloacae ATCC 13047, and effective cholesterol removal ability in vitro. Treatment with heat-killed HOM2217 significantly reduced lipid accumulation and intracellular triglyceride production in 3T3-L1 adipocytes. Daily treatment of HFD-fed rats with HOM2217 for 7 weeks decreased body weight, body weight gain, and body fat without changes in food intake. HOM2217 also significantly increased the serum high-density lipoprotein cholesterol (HDL-C) level, decreased the serum tumor necrosis factor (TNF-a) and increased short-chain fatty acid (SCFA) (formic acid, acetic acid, and butyric acid) levels in the cecum. Thus, HOM2217 could potentially prevent obesity in rats by inhibiting inflammatory responses and regulating lipid metabolism and SCFAs expression. Therefore, HOM2217 has potential as an alternative treatment for obesity. [ABSTRACT FROM AUTHOR]

Objectives: To investigate the effects of low‐intensity pulsed ultrasound (LIPUS) on the proliferation, differentiation, and tumor necrosis factor‐α (TNF‐α)‐induced lipolysis of 3T3‐L1 cells, and to explore the feasibility of regulating the release of free fatty acids (FFA) to prevent lipotoxicity. Methods: Different intensities (30, 60, 90, and 120 mW/cm2) of LIPUS were applied to 3T3‐L1 preadipocytes for different durations (5, 10, 15, 20, 25, and 30 minutes). Appropriate parameters for subsequent experiments were selected by assessing cell viability. The effect of LIPUS on the proliferation and differentiation of 3T3‐L1 cells was evaluated by microscope observation, flow cytometry, and lipid content determination. After treated with LIPUS and TNF‐α (50 ng/mL), the degree of lipolysis was assessed by measuring the extracellular FFA content. Quantitative real‐time polymerase chain reaction (qRT‐PCR) was used to detect the mRNA expression of relevant genes. Results: Different parameters of LIPUS significantly enhance the viability of 3T3‐L1 cells (P <.05), with 20 minutes and 30 mW/cm2 as the most suitable settings. After LIPUS treatment, 3T3‐L1 cell proliferation accelerated, apoptosis rate and G1 phase cell proportion decreased, the content of lipid droplets and TG was increased in differentiated cells, while FFA release decreased (P <.05). The expression of PCNA, PPARγ, C/EBPα, Perilipin A mRNA increased, and the expression of TNF‐α, ATGL, HSL mRNA decreased (P <.05). Conclusions: LIPUS could promote the proliferation and differentiation of 3T3‐L1 cells and inhibit TNF‐α‐induced lipolysis, indicating its potential as a therapy for mitigating lipotoxicity caused by decompensated adipocytes. [ABSTRACT FROM AUTHOR]

Objective: The study was designed to test solvent extracts of Rubus suavissimus (RS) for compounds that regulate lipid metabolism and to measure their relative activity as quantitative markers of lipid metabolism-regulating activities based on the spectrum-effect relationship. Method: RS was extracted with 95% ethanol, the different extraction parts were successively extracted with petroleum ether, methylene chloride, ethyl acetate and n-butanol, respectively. The solution of different extraction parts was applied to the induced differentiated 3T3-L1 preadipocytes, and the oil red O staining and triglyceride release from the cells were determined to screen the best metabolism-regulating site. Analysis of different extraction sited by UPLC-Q-TOF-MS/MS technique to separate and identify the components. Based on specific assays, image intensity analysis, grey correlation and partial least squares regressions, the spectrum-effect relationship of the most abundant active components and their ability to regulate lipid metabolism were determined, and the quality markers were screened. Results: Ethyl acetate and n-butanol extraction site was the best metabolism-regulating site. Four components, including ellagic acid, centaurin-3-O-rutinoside, rubusoside and enantio-kauri-16-ene-19-carboxylic acid-13-O-β-D-glucoside were highly associated with metabolism-regulating effects. It could be used as quality markers for regulating lipid metabolism. Conclusion: Investigation of the quality markers of RS extracts based on the spectrum-effect relationship is of great importance for elucidating the pharmacodynamics, screening the core quality markers for therapeutic activity, and ensuring the safety and rational application of traditional medicines. [ABSTRACT FROM AUTHOR]

Interleukin-33 (IL-33), an emerging cytokine within the IL-1 family, assumes a pivotal function in the control of obesity. However, the specific mechanism of its regulation of obesity formation remains unclear. In this study, we found that the expression level of IL-33 increased in visceral adipose tissue in mice fed with a high-fat diet (HFD) compared with that in mice fed with a normal diet (ND). In vitro, we also found the expression level of IL-33 was upregulated during the adipogenesis of 3T3-L1 cells. Functional test results showed that knockdown of IL-33 in 3T3-L1 cells differentiation could promote the accumulation of lipid droplets, the content of triglyceride and the expression of adipogenic–related genes (i.e. PPAR-γ, C/EBPα, FABP4, LPL, Adipoq and CD36). In contrast, overexpression of IL-33 inhibits adipogenic differentiation. Meanwhile, the above tests were repeated after over-differentiation of 3T3-L1 cells induced by oleic acid, and the results showed that IL-33 played a more significant role in the regulation of adipogenesis. To explore the mechanism, transcriptome sequencing was performed and results showed that IL-33 regulated the PPAR signaling pathway in 3T3-L1 cells. Further, Western blot and confocal microscopy showed that the inhibition of IL-33 could promote PPAR-γ expression by inhibiting the Wnt/β-catenin signal in 3T3-L1 cells. This study demonstrated that IL-33 was an important regulator of preadipocyte differentiation and inhibited adipogenesis by regulating the Wnt/β-catenin/PPAR-γ signaling pathway, which provided a new insight for further research on IL-33 as a new intervention target for metabolic disorders. [ABSTRACT FROM AUTHOR]

Objective: To investigate the active components, protein targets, and mechanisms underlying the anti-obesity effects of Hippophae fructus using network pharmacology and molecular docking techniques, and to validate its in vitro anti-obesity efficacy. Methods: The TCMSP platform was utilized to retrieve the active components and targets of Hippophae fructus, and disease targets were collected. Venny 2.0.2 was used to identify the intersection of targets between Hippophae fructus and obesity-related targets. The STRING database was used to establish a drug-target-disease protein interaction (PPI) network. The intersecting targets were analyzed using the David database to perform GO enrichment analysis and KEGG pathway analysis. Cytoscape 3.9.1 was used to construct a network diagram of the components of Hippophae fructus, anti-obesity targets, and related signaling pathways. Autodock Dock 1.5.7 and Pymol 2.2.0 were used to carry out molecular docking between the core targets of Hippophae fructus and its components, followed by visualization. The in vitro anti-obesity effect of Hippophae fructus extract was evaluated through cell experiments using 3T3-L1 cells. Results: A total of 33 active components, 2820 disease targets, and 151 intersection targets of Hippophae fructus were identified. The main active components included flavonoids, vitamins, and sterols, while key targets involved AKT1, TNF, IL6, TP53, VEGFA, CASP3, and others. KEGG pathway enrichment analysis revealed 131 signaling pathways, including those related to malignant tumors, lipid and atherosclerosis, and AGE-RAGE signaling. Molecular docking results demonstrated favorable binding interactions between the core targets and the corresponding active components of Hippophae fructus. The in vitro experiments indicated that Hippophae fructus extract exhibited inhibitory effects on the proliferation of 3T3-L1 pre-adipocytes. Conclusion: This study reveals that Hippophae fructus exerts antiobesity effects through multiple components, targets, and pathways, providing valuable insights for its clinical research and product development. [ABSTRACT FROM AUTHOR]

Obesity is the excessive accumulation of body fat resulting from impairment in energy balance mechanisms. In this study, we aimed to investigate the mechanism whereby GABA (γ-aminobutyric acid) prevents high-fat diet-induced obesity, and whether it induces lipolysis and browning in white adipose tissue (WAT), using high-fat diet (HFD)-fed obese mice and 3T3-L1 adipocytes. We demonstrated that GABA substantially inhibits the body mass gain of mice by suppressing adipogenesis and lipogenesis. Consistent with this result, histological analysis of WAT demonstrated that GABA decreases adipocyte size. Moreover, we show that GABA administration decreases fasting blood glucose and improves serum lipid profiles and hepatic lipogenesis in HFD-fed obese mice. Furthermore, Western blot and immunofluorescence analyses showed that GABA activates protein kinase A (PKA) signaling pathways that increase lipolysis and promote uncoupling protein 1 (UCP1)-mediated WAT browning. Overall, these results suggest that GABA exerts an anti-obesity effect via the regulation of lipid metabolism. [ABSTRACT FROM AUTHOR]

Fat browning, which converts white adipose tissue to brown, has attracted attention as a promising strategy for the treatment of obesity. Betanin (BT) has been reported to have potential anti-obesity activity. 3T3-L1 cells were differentiated for 7 days during BT treatment. The BT concentration range for the study was determined using an MTT assay, and lipid accumulation was evaluated by Oil-Red-O staining. The expression of protein level was analyzed by Western blot. Immunofluorescence images were performed with confocal microscopy to visually show the amount and location of thermogenesis factor uncoupling protein1 (UCP1) and mitochondria. qRT-PCR was performed to evaluate mRNA expression. BT inhibited lipid accumulation and increased the expression of UCP1, peroxisome-proliferator-activated receptor gamma (PPARγ), and PPARγ coactivator-1 alpha (PGC-1α). In addition, the increases in beige adipocyte-specific markers were observed, supporting BT-mediated browning of the fat tissue. The UCP1 was localized in the inner membrane of the mitochondria, and its expression was associated with mitochondrial activation. Consistent with this, the mRNA expression of mitochondrial biogenesis markers increased in 3T3-L1 cells after BT treatment. Immunofluorescence staining also indicated an increased number of mitochondria and UCP1, respectively. Moreover, BT inhibited lipogenesis and enhanced lipolysis and fatty acid oxidation. This mechanism has been suggested to be mediated by an adenosine monophosphate-activated protein kinase (AMPK) pathway. BT induces fat browning and regulates lipid metabolism via the AMPK-mediated pathway in 3T3-L1 cells, suggesting that BT can be a promising candidate for controlling obesity. [ABSTRACT FROM AUTHOR]

Natural polyphenol-rich plant resources, such as agricultural waste, were proven to diminish insulin resistance and weight gain in rats on a high-fat diet. To test whether date seed polyphenol pills (DSPPs) might lower adipose tissue accumulation by precisely affecting adipocytes, we explored the impacts of DSPPs on cell proliferation, differentiation, and lipolysis in 3T3-L1 cells. We utilized tablets made commercially from date seed polyphenols that were mostly composed of epicatechin (45.9 g/kg). The total polyphenol and antioxidant capacities of the digested and non-digested DSPPs were also evaluated. DSPPs at doses of 25, 50, and 100 µg/mL hindered the proliferation of both pre-confluent preadipocytes and mature post-confluent adipocytes. DSPPs decreased the quantity of viable cells in completely developed adipocytes. Treatment with 100 µg/mL of DSPPs decreased the basal lipolysis of completely differentiated adipocytes but modestly boosted epinephrine-induced lipolysis. A significant transcription factor for the adipogenic gene, the peroxisome proliferator-activated receptor (PPAR), was repressed by DSPPs, which significantly decreased lipid buildup. The total polyphenol and antioxidant capacities were also increased after digestion with a good bubble Pearson correlation between both. DSPPs may have anti-obesity and anti-diabetic characteristics by inhibiting adipocyte development and basal lipolysis, which could be commercially industrialized. [ABSTRACT FROM AUTHOR]

Background and Objectives: This study evaluated the in vitro anti-adipogenic and anti-inflammatory properties of black cumin (Nigella sativa L.) seed extract (BCS extract) as a potential candidate for developing herbal formulations targeting metabolic disorders. Materials and Methods: We evaluated the BCS extract by assessing its 2,2-diphenyl-1-picrohydrazyl (DPPH) radical scavenging activity, levels of prostaglandin E2 (PGE2) and nitric oxide (NO), and mRNA expression levels of key pro-inflammatory mediators. We also quantified the phosphorylation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB) and mitogen-activated protein kinases (MAPK) signaling molecules. To assess anti-adipogenic effects, we used differentiated 3T3-L1 cells and BCS extract in doses from 10 to 100 μg/mL. We also determined mRNA levels of key adipogenic genes, including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/BEPα), adipocyte protein 2 (aP2), lipoprotein lipase (LPL), fatty acid synthase (FAS), and sterol-regulated element-binding protein 1c (SREBP-1c) using real-time quantitative polymerase chain reaction (qPCR). Results: This study showed a concentration-dependent DPPH radical scavenging activity and no toxicity at concentrations up to 30 μg/mL in Raw264.7 cells. BCS extract showed an IC50 of 328.77 ± 20.52 μg/mL. Notably, pre-treatment with BCS extract (30 μg/mL) significantly enhanced cell viability in lipopolysaccharide (LPS)-treated Raw264.7 cells. BCS extract treatment effectively inhibited LPS-induced production of PGE2 and NO, as well as the expression of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS), interleukin (IL)-1β and IL-6, possibly by limiting the phosphorylation of p38, p65, inhibitory κBα (I-κBα), and c-Jun N-terminal kinase (JNK). It also significantly attenuated lipid accumulation and key adipogenic genes in 3T3-L1 cells. Conclusions: This study highlights the in vitro anti-adipogenic and anti-inflammatory potential of BCS extract, underscoring its potential as a promising candidate for managing metabolic disorders. [ABSTRACT FROM AUTHOR]

Summary: High amounts of raspberries are wasted every year, which are rich source of bioactive compounds. Thus, raspberry waste was used to obtain the essential oil by maceration and the oleoresin by supercritical fluid extraction (SFE). Higher amounts of total phenolics (185 mg GAE g−1) and flavonoids (11.0 mg QE g−1) were obtained in oleoresins than in essential oil (131 mg GAE g−1 and 9.0 mg QE g−1, respectively). DPPH radical scavenging was 1350 μm TE g−1 for oleoresins and 890 μm TE g−1 for essential oil. SFE allowed the extraction of compounds not reported before in red raspberry such as a pinocembrin and farnesol. Oleoresin at 20 μg mL−1 induced a decrease in lipid accumulation during the 3T3‐L1 cell differentiation process, but no after cells were differentiated. Red raspberry waste is an important source of bioactives that can be extracted using green technologies to generate high commercial value by‐products. [ABSTRACT FROM AUTHOR]

Background: Gremlin-1 (GREM1) and Gremlin-2 (GREM2) are bone morphogenetic protein antagonists that play important roles in organogenesis, tissue differentiation, and tissue homeostasis. Although GREM1 has been reported to be involved in promoting various cancers, little has been reported about effects of GREM2 on cancer. Recently, it has been reported that GREM2 can inhibit adipogenesis in adipose-derived stromal/stem cells. However, as an inhibitor of adipogenesis, the role of GREM2 in cancer progression is not well understood yet. Methods: Pre-adipocyte 3T3-L1 cells overexpressing mock or Grem2 were established using a lentiviral transduction system and differentiated into adipocytes-mock and adipocytes-Grem2, respectively. To investigate the effect of adipocyte-Grem2 on breast cancer cells, we analyzed the proliferative and invasion abilities of spheroids using a 3D co-culture system of breast cancer cells and adipocytes or conditioned medium (CM) of adipocytes. An orthotopic breast cancer mouse model was used to examine the role of adipocytes-Grem2 in breast cancer progression. Results: Grem2 overexpression suppressed adipogenesis of 3T3-L1 cells. Proliferative and invasion abilities of spheroids formed by co-culturing MTV/TM-011 breast cancer cells and adipocytes-Grem2 were significantly reduced compared to those of spheroids formed by co-culturing MTV/TM-011 cells and adipocytes-mock. Compared to adipocytes-mock, adipocytes-Grem2 showed decreased mRNA expression of several adipokines, notably IL-6. The concentration of IL-6 in the CM of these cells was also decreased. Proliferative and invasive abilities of breast cancer cells reduced by adipocytes-Grem2 were restored by IL-6 treatment. Expression levels of vimentin, slug, and twist1 in breast cancer cells were decreased by treatment with CM of adipocytes-Grem2 but increased by IL-6 treatment. In orthotopic breast cancer mouse model, mice injected with both MTV/TM-011 cells and adipocytes-Grem2 showed smaller primary tumors and lower lung metastasis than controls. However, IL-6 administration increased both the size of primary tumor and the number of metastatic lung lesions, which were reduced by adipocytes-Grem2. Conclusions: Our study suggests that GREM2 overexpression in adipocytes can inhibit adipogenesis, reduce the expression and secretion of several adipokines, including IL-6, and ultimately inhibit breast cancer progression. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Immunology is the property of Medical Periodical Society of Jilin and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The present study was aimed to evaluate the anti-adipogenic activity of piperine (PIP) and epigallocatechin gallate (EGCG) in 3T3-L1 cells. In cytotoxicity studies, PIP and EGCG showed IC50 values of 260 and 218 µM respectively and in combination (20 µM each) did not show cytotoxicity. Treatment with PIP and EGCG (20 µM each) significantly (p<.01) inhibited cell differentiation, lipid droplets deposition and enhanced glycerol release in 3T3-L1 cells. The secreted level of leptin was decreased but adiponectin level was increased in treated 3T3-L1 cells than untreated cells. In molecular expression studies, key adipogenic genes PPAR-γ, SREBP-1c, FAS, Fab-4, C/EBP-α and HMG-CoA reductase were markedly down-regulated but UCP-1 was up-regulated in treated 3T3-L1 cells and the same trend was observed in expression levels of selected proteins. In conclusion, our results demonstrated a combination of PIP and EGCG exhibited strong anti-adipogenic and lipid lowering effect than individual treatments due to synergism. [ABSTRACT FROM AUTHOR]

The signaling pathway of fatty acids in the context of obesity is an extensively explored topic, yet their primary mechanism of action remains incompletely understood. This study aims to examine the effect of docosahexaenoic acid (DHA) on some crucial aspects of adipogenesis in differentiating 3T3-L1 cells, using palmitic acid-treated (PA), standard differentiated, and undifferentiated adipocytes as controls. Employing 60 µM DHA or PA, 3T3-L1 preadipocytes were treated from the onset of adipogenesis, with negative and positive controls included. After eight days, we performed microscopic observations, cell viability assays, the determination of adiponectin concentration, intracellular lipid accumulation, and gene expression analysis. Our findings demonstrated that DHA inhibits adipogenesis, lipolysis, and glucose uptake by suppressing peroxisome proliferator-activated receptor gamma (Pparg) and G-protein coupled receptor 120 (Gpr120) gene expression. Cell cytotoxicity was ruled out as a causative factor, and β-oxidation involvement was suspected. These results challenge the conventional belief that omega-3 fatty acids, acting as Pparg and Gpr120 agonists, promote adipogenesis and enhance insulin-dependent glucose cell flux. Moreover, we propose a novel hypothesis suggesting the key role of the co-repressor G protein pathway suppressor 2 in mediating this process. Additional investigations are required to elucidate the molecular mechanisms driving DHA's anti-adipogenic effect and its broader health implications. [ABSTRACT FROM AUTHOR]

Obesity is a major risk factor for a variety of diseases and contributes to chronic inflammation. Resveratrol is a naturally occurring antioxidant that can reduce adipogenesis. In this study, the antiadipogenic and anti-inflammatory activities of resveratrol-enriched rice were investigated in 3T3-L1 adipocyte cells. Cotreatment of dexamethasone and isobutylmethylxanthin upregulated adipogenic transcription factors and signaling pathways. Subsequent treatment of adipocytes with rice seed extracts suppressed the differentiation of 3T3-L1 by downregulating adipogenic transcription factors (peroxisome proliferator-activated receptor γ and CCAAT/enhancer-binding protein α) and signaling pathways (extracellular signal-regulated kinase 1/2 and protein kinase B Akt), this was especially observed in cells treated with germinated resveratrol-enriched rice seed extract (DJ526_5). DJ526_5 treatment also markedly reduced lipid accumulation in the cells and expression of adipogenic genes. Lipopolysaccharide (LPS)-induced inflammatory cytokines (prostaglandin-endoperoxide synthase 2 (COX-2), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6) decreased in cells treated with DJ526_5. Collectively, DJ526_5 exerts antiadipogenic effects by suppressing the expression of adipogenesis transcription factors. Moreover, DJ526_5 ameliorates anti-inflammatory effects in 3T3-L1 adipocytes by inhibiting the activation of phosphorylation NF-κB p65 and ERK ½ (MAPK). These results highlight the potential of resveratrol-enriched rice as an alternative obesity-reducing and anti-inflammatory agent. [ABSTRACT FROM AUTHOR]

Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox regulation. The redox activity of APE1/Ref-1 is involved in inflammatory responses and regulation of DNA binding of transcription factors related to cell survival pathways. However, the effect of APE1/Ref-1 on adipogenic transcription factor regulation remains unknown. In this study, we investigated the effect of APE1/Ref-1 on the regulation of adipocyte differentiation in 3T3-L1 cells. During adipocyte differentiation, APE1/Ref-1 expression significantly decreased with the increased expression of adipogenic transcription factors such as CCAAT/enhancer binding protein (C/EBP)-α and peroxisome proliferator-activated receptor (PPAR)-γ, and the adipocyte differentiation marker adipocyte protein 2 (aP2) in a time-dependent manner. However, APE1/Ref-1 overexpression inhibited C/EBP-α, PPAR-γ, and aP2 expression, which was upregulated during adipocyte differentiation. In contrast, silencing APE1/Ref-1 or redox inhibition of APE1/Ref-1 using E3330 increased the mRNA and protein levels of C/EBP-α, PPAR-γ, and aP2 during adipocyte differentiation. These results suggest that APE1/Ref-1 inhibits adipocyte differentiation by regulating adipogenic transcription factors, suggesting that APE1/Ref-1 is a potential therapeutic target for regulating adipocyte differentiation. [ABSTRACT FROM AUTHOR]

Obesity is a disease in which fat is abnormally or excessively accumulated in the body, and many studies have been conducted to overcome it with various techniques. In this study, we evaluated whether micro-current stimulation (MCS) can be applied to prevent obesity by regulating the adipogenesis through 3T3-L1 cells and ob/ob mice. To specify the intensity of MCS, Oil Red O staining was conducted with various intensities of MCS. Based on these, subsequent experiments used 200 and 400 μA for the intensity of MCS. The expressions of insulin signaling pathway-related proteins, including phosphorylation of IGF-1 and IR, were decreased in all MCS groups, and in turn, downstream signals such as Akt and ERK were decreased. In addition, MCS reduced the nucleus translocation of PPAR-γ and decreased the protein expression of C/EBP-α. In the ob/ob mouse model, MCS reduced body weight gain and abdominal adipose tissue volume. In particular, the concentration of triglycerides in serum was also decreased. Taken together, our findings showed that MCS inhibited lipid accumulation by regulating insulin signaling in 3T3-L1, and it was effective at reducing body weight and adipose tissue volume in ob/ob mice. These suggest that MCS may be a useful treatment approach for obesity. [ABSTRACT FROM AUTHOR]

Polycyclic aromatic hydrocarbons (PAHs) are one of the most prevalent classes of environmental pollutants. Some evidence shows that PAHs could be involved in human obesity. However, little is known about the distribution patterns of PAHs in human adipose tissue (AT) and the role of PAHs on adipogenesis/lipogenesis. The aims of this pilot study were to determine concentrations of 16 PAHs defined as high-priority pollutants in the plasma and adipose tissue of French and Polish bariatric patients, as well as their correlation with body mass index (BMI), plasma and AT adipokines expression levels. We finally investigated the role of naphthalene on cell proliferation, viability, and differentiation in 3T3-L1 preadipocytes. The concentration of most PAHs was similar in the three types of AT and it was significantly higher in AT as compared to plasma, suggesting bioaccumulation. Polish patients had higher PAH levels in AT than French ones. Only the concentration of naphthalene in AT was positively correlated with the BMI and serum or adipose chemerin, adiponectin and resistin expression, in French but not in Polish patients, who had significantly higher BMIs. Moreover, naphthalene exposure increased the cell proliferation of 3T3-L1 preadipocytes and lipogenesis, and increased the expression of genes involved in adipogenesis after cell differentiation. Taken together, PAHs and more particularly naphthalene could be an obesogenic molecule and increase the risk of obesity. [ABSTRACT FROM AUTHOR]

The present study explores the molecular mechanism of anti-obesity effect of total lutein oxidized products (LOPs) in high-fat diet-induced obese C57BL/6 mice. Total LOPs (50, 100, 200 mg/kg b.wt.) were intubated for 22 weeks. Lipid profile, peroxides, AST, ALT, GST activities, lipid metabolizing enzymes/molecules, 3T3-L1 cell differentiation, TGF-β1 and p38 MAPK expressions were studied. Total LOPs at 100 mg/kg b.wt. reduced plasma and hepatic cholesterol, triglycerides, phospholipids, ALT and AST activities. FAS and CPT activities were inhibited in the liver homogenate, and epididymal WAT (eWAT). Total LOPs at 200 mg/kg b.wt. reduced HMGCoA:Mevalonate in eWAT, lipid peroxides in eWAT and liver. GST activity was high in the liver and eWAT. Total LOPs (100μg/ml) suppressed adipogenesis and decreased the accumulation of lipid droplets from two to eight days. Adiponectin was upregulated, PPAR γ, lipoprotein lipase, TGF-β1, and p38 MAPK were down-regulated. The results have shown that total LOPs are potent anti-obese molecules. ABBREVIATIONS: AIN 93G: American institute of nutrition 93 for growth; ALT: alanine transaminase; AST: aspartate transaminase; CEBPα: ccaat/enhancer binding protein α; C-DNA: complementary deoxyribonucleic acid; CPT: carnitine palmitoyl transferase; GST: glutathione-S-transferase; ELISA: enzyme-linked immune sorbent assay; FAS: fatty acid synthase; HDL: high-density lipoprotein; HFD: high-fat diet; HMG CoA: β-Hydroxy β-methylglutaryl-Coenzyme A; LDL: low-density lipoprotein; LOPs: total lutein oxidized products; LPL: lipoprotein lipase; MDA-Malonaldehyde; MTT: 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyl tetrazolium bromide; MVA: mevalonate; p38MAPK: p38 mitogen-activated protein kinases; RNA: ribonucleic acid; RTq-PCR: quantitative reverse transcription polymerase chain reaction; PPAR-γ: peroxisome proliferator-activated receptor-gamma; TGF-β1: transforming growth factor beta 1; WAT: white adipose tissue. [ABSTRACT FROM AUTHOR]

The study aimed to compare the effect of 100 µM docosahexaenoic acid and oleic acid alone, and as a combination in immature and already mature 3T3-L1. The cells were subjected to adipogenic induction for 2 days and then were maintained in a high glucose culture medium supplemented with 10 µg/mL of insulin for additional 16 days. The treatment with docosa-hexaenoic acid and oleic acid lasted 9 days in total, as half of the cells were treated during induction (from day 1 to 9), and the rest--after reaching maturity (day 9--18). The palmitic fatty acid was used as a positive control. Applied in immature adipocytes, docosahexaenoic acid prevented intracellular neutral lipid deposition and reduced lipolysis upon well-preserved glucose utilization. At the same time, oleic acid alone or combined with docosahexaenoic acid increased lipid accumulation and slightly attenuated lipolysis only related to the palmitic fatty acid-treated group. However, free fatty acids application in mature adipocytes did not cause any beneficial effect. They elevated lipolysis rate compared to the induced control, which in vivo might be a predisposing factor for ectopic fat accumulation. The results emphasized that free fatty acids' impact on adipocyte metabolism was highly affected by their maturation stage and should be considered in anti-obesity strategies based on free fatty acids signaling modulations. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Salix pseudolasiogyne (Salicaceae), the "weeping willow," has been used in traditional Korean medicine to treat pain and fever due to its high concentrations of salicylic acid and salicin. The present study investigated bioactive compounds from S. pseudolasiogyne twigs to discover bioactive natural products. Phytochemical investigation of the ethanol (EtOH) extract of S. pseudolasiogyne twigs followed by liquid chromatography–mass spectrometry (LC/MS)-based analysis led to the isolation of two salicin derivatives, salicortinol and salicortin, the structures of which were determined by interpretation of their NMR spectra and data from the LC/MS analysis. To the best of our knowledge, this is the first report of salicortinol isolated from S. pseudolasiogyne. The isolated compounds were evaluated for their anti-adipogenic effects in 3T3-L1 cells. Both salicortinol and salicortin were found to significantly inhibit adipocyte differentiation in 3T3-L1 cells. In particular, salicortin exhibited a strong inhibitory effect on lipid accumulation. Furthermore, salicortin inhibited the expression of lipogenic and adipogenic transcription factors, including FASN, FABP4, C/EBPα, C/EBPβ, and PPARγ, without inducing cytotoxicity. These results suggest that salicortin could be a potential therapeutic compound for the prevention or treatment of metabolic disorders such as obesity. [ABSTRACT FROM AUTHOR]

Due to its propensity to boost energy expenditure, browning of white fat is emerging as an intriguing and prospective target for therapeutic intervention in obesity. Here, we report that L-dihydroxyphenylalanine (L-Dopa), used as a gold standard therapy in Parkinson's disease, induces browning in 3T3-L1 adipocytes by increasing the expression levels of beige-specific marker genes such as Cd137, Cited1, Cidea, Tbx1, Prdm16, and Ucp1. In addition, exposure to L-Dopa induces a remarkable increase in the expressions of proteins involved in thermogenesis in white adipocytes. L-Dopa treatment also regulates 3T3-L1 adipocytes by markedly increasing protein expressions of p-AMPK, p-HSL, CPT1, ACOX1, and PPARα while decreasing FAS, ACC, C/EBPα, and PPARγ, suggesting enhanced lipolysis and fatty acid oxidation as well as reduced lipogenesis and adipogenesis, respectively. Molecular docking studies elucidated that L-Dopa binds to dopamine receptor D1 (DRD1) and β3-AR, thereby predicting the potential receptor candidates that activate protein kinase A (PKA), the master regulator of lipid metabolism. Mechanistic studies indicate that the browning potential of L-Dopa in 3T3-L1 white adipocytes is mediated by DRD1 and β3-AR activation, which consequently stimulates the PKA/p38 MAPK/ERK signaling pathway. In conclusion, L-Dopa appears to be a promising therapeutic candidate in the fight against obesity due to its inherent role in the browning of 3T3-L1 adipocytes via both the dopaminergic and adrenergic pathways. To our knowledge, this is the first report that demonstrates the browning potential of L-Dopa in white adipocytes. Our results may assist to expand the understanding on the contradictory findings in literature, related to the association between L-Dopa and weight loss observed in Parkinson's disease patients. [ABSTRACT FROM AUTHOR]

We investigated the effects of derhamnosylmaysin (DM) on adipogenesis and lipid accumulation in 3T3-L1 adipocytes. Our data showed that DM inhibited lipid accumulation and adipocyte differentiation in 3T3-L1 cells. Treatment of 3T3-L1 adipocytes with DM decreased the expression of major transcription factors, such as sterol regulatory element-binding protein-1c (SREBP-1c), the CCAAT-enhancer-binding protein (CEBP) family, and peroxisome proliferator-activated receptor gamma (PPARγ), in the regulation of adipocyte differentiation. Moreover, the expression of their downstream target genes related to adipogenesis and lipogenesis, including adipocyte fatty acid-binding protein (aP2), lipoprotein lipase (LPL), stearyl-CoA-desaturase-1 (SCD-1), acetyl-CoA carboxylase (ACC), and fatty acid synthase (FAS), was also decreased by treatment with DM during adipogenesis. Additionally, DM attenuated insulin-stimulated phosphorylation of Akt. These results first demonstrated that DM inhibited adipogenesis and lipogenesis through downregulation of the key adipogenic transcription factors SREBP-1c, the CEBP family, and PPARγ and inactivation of the major adipogenesis signaling factor Akt, which is intermediated in insulin. These studies demonstrated that DM is a new bioactive compound for antiadipogenic reagents for controlling overweight and obesity. [ABSTRACT FROM AUTHOR]

Background: MicroRNAs (MiRNAs) are known to participate in preadipocyte differentiation, but the manner in which miR-146a-5p participates in this process remains unclear. This study was performed to examine the participation of miR-146a-5p in 3T3-L1 cell differentiation. Material and Methods: miR-146a-5p expression was upregulated and down-regulated to examine effects on 3T3-L1 cell differentiation. Bioinformatics analysis was performed to predict its target genes, and the signaling pathway it regulates was identified by qRT-PCR and Western blotting. The expression of miR-146a-5p in epididymal adipose tissue from obese mice and in an obese mouse adipose cell model was examined by qRT-PCR. Results: 3T3-L1 cells differentiated into mature adipocytes successfully, as verified by increased areas of intracellular lipid droplets and elevated expression of mature adipocyte markers, and these cells had elevated miR-146a-5p expression. The intracellular lipid droplet and triglyceride contents and the expression of mature adipocyte markers were significantly increased in miR-146a-5p–overexpressing 3T3-L1 cells and markedly decreased in miR-146a-5p–inhibited 3T3-L1 cells. ErbB4 was a predicted target gene of miR-146a-5p. In miR-146a-5p–overexpressing 3T3-L1 cells, ErbB4 expression and ERK1/2 phosphorylation were decreased and the expression of PPAR-γ was increased; the opposite was observed in miR-146a-5p–inhibited 3T3-L1 cells. In addition, miR-146a-5p expression was significantly increased in the mouse epididymal adipose tissue and adipose cell model. Conclusions: Upregulated miR-146a-5p expression was related to 3T3-L1 cell differentiation. MiR-146a-5p promoted 3T3-L1 cell differentiation by targeting ErbB4 and via the ERK1/2/PPAR-γ signaling pathway. [ABSTRACT FROM AUTHOR]

Isoacteoside is a caffeoyl phenylethanoide glycoside found in various plant parts, such as the flower of Magnolia denudata. In particular, magnolia has been studied for its anti-obesity, anticancer, and anti-inflammatory effects. However, isoacteoside has not been extensively studied, except for its anti-inflammatory effects. In this study, the anti-obesity effects of isoacteoside were investigated in 3T3-L1 mouse pre-adipocytes. Isoacteoside treatment did not induce cytotoxicity in 3T3-L1 cells up to a concentration of 1000 μM. The anti-obesity effects on 3T3-L1 cells were confirmed using oil red O staining. In addition, the expression of obesity-related proteins and genes, such as peroxisome proliferator-activated acceptor gamma (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBPα), and perilipin (PLIN1), was determined by western blotting and qRT-PCR assays to confirm the anti-obesity effects of isoacteoside. The results of this study suggest that isoacteoside, a natural substance isolated from plant extracts, is not highly toxic to normal cells, such as pre-adipocytes, and displays anti-obesity effects in vitro. [ABSTRACT FROM AUTHOR]

Copyright of Tianjin Medical Journal is the property of Tianjin Medical Journal and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Blue honeysuckle (BH; Lonicera caerulea L.), which is a traditional medicinal plant, is known to be a rich source of anthocyanins and phenolic acids due to its strong antioxidant and anti-inflammatory activities. Its anti-obesity effects, which are a result of attenuating abnormal lipid and glucose metabolisms, have also been reported. Aim: The purpose of this study is to investigate the effect of BH on genes and proteins that are involved in the adipocyte differentiation using 3T3-L1 cells. Methods: The effects of the water extracts of the BH were examined on adipogenesis and lipolysis using a biochemical and molecular analysis of the 3T3-L1 cells. Results: Cyanidin-3-O-glucoside (C3G) from the BH extract was determined in order to contain 1.67 mg/g by the high-performance liquid chromatography analysis. The lipid accumulation in the adipocytes was reduced, which ranged from 58 to 26% in the BH (500 and 1,000 µg/mL) compared to the control group. The lipolysis that was measured by the glycerol content was not affected by the BH at 1,000 μg/mL. The BH downregulated the expression of the main transcription factors of the adipogenesis pathway, such as peroxisome proliferator-activated receptor γ, 1, 2, adipose differentiation-related protein, CCAAT/enhancer binding protein α, and acetyl CoA carboxylase, while increasing the expression of the Adenosine monophosphate (AMP)-activated protein kinase α. Conclusion: These findings suggest that the BH is a good source of C3G, and it could be effective in regard to inhibiting the adipogenesis as opposed to the lipolysis, which indicates the potential for natural anti-obesity ingredients. [ABSTRACT FROM AUTHOR]

Culturing cells on bio-gels are believed to provide a more in vivo-like extracellular matrix. 3T3-L1 cells cultured on Matrigel® significantly alteregd their proliferation and differentiation as compared to growth on tissue culture-coated polystyrene surfaces. Growth on a 250-μm thick layer of Matrigel® facilitated the formation of cellular aggregates of 3T3-L1 cells. Differentiation of 3T3-L1 cells cultured on Matrigel® demonstrated increased levels of mRNA levels for key adipogenic transcription factors (PPARγ, C/EBPα, SREBP1), lipogenic markers (FAS, FABP4, LPL, PLIN1) and markers of adipocyte maturity (LEP), compared to cells cultured directly on a polystyrene tissue culture surface. The gene expression of extracellular matrix proteins (FN1, COL1A1, COL4A1, COL6, LAM) was decreased in 3T3-L1 cells cultured on Matrigel®. Furthermore, growth on Matrigel® increased lipid accumulation in 3T3-L1 cells in the presence and absence of rosiglitazone, a thiazolidinedione routinely used to optimize differentiation in these cells. These changes in adipocyte gene expression and lipid accumulation patterns may be a result of the increased cell–cell and cell–ECM interactions occurring on the Matrigel®, a scenario that is more reflective of an in vivo model. Taken together, our data advance the understanding of the value of culturing 3T3-L1 cells on Matrigel®. [ABSTRACT FROM AUTHOR]

In the search for new drugs against obesity, the chronic disease that threatens human health worldwide, several works have focused on the study of estrogen homologs because of the role of estrogen receptors (ERs) in adipocyte growth. The isoflavone equol, an ERβ agonist, has shown beneficial metabolic effects in in vivo and in vitro assays; however, additional studies are required to better characterize its potential for body weight control. Here, we showed that the treatment of 3T3-L1 cells with 10 μM of S-equol for the first three days of the adipocyte differentiation protocol was able to prevent cells becoming semi-rounded and having a lipid droplet formation until the seventh day of culture; moreover, lipid accumulation was reduced by about 50%. Congruently, S-equol induced a reduction in mRNA expression of the adipogenic markers C/EBPα and PPARγ, and adipokines secretion, mainly Adiponectin, Leptin, Resistin, and MCP-1, while the release of PAI-1 was augmented. Moreover, it also reduced the expression of ERα and attenuated the subexpression of ERβ associated with adipogenesis. Altogether, our data suggested that S-equol binding to ERβ affects the transcriptional program that regulates adipogenesis and alters adipocyte functions. Future efforts will focus on studying the impact of S-equol on ER signaling pathways. [ABSTRACT FROM AUTHOR]

Obesity is caused by excessive fat cells and the overgrowth of adipocytes and is a major risk factor for several chronic illnesses. Aronia melanocarpa fruit is rich in anthocyanins and polyphenols and has protective effects against various diseases. In this study, we examined the effect of Aronia extract (Aronia bioactive fraction, ABF®) on the biomarkers of the adipogenic pathway during adipocyte differentiation of 3T3-L1 cells. Lipid accumulation was verified by Oil Red O staining. mRNA and protein expression of lipoprotein lipase (LPL), CCAAT/enhancer-binding protein α (C/EBPα), peroxisome proliferator-activated receptor γ (PPARγ), fatty acid-binding protein 2 (FABP2), and fatty acid synthase (FAS) were assayed by RT-qPCR and Western blot analyses. Adiponectin and leptin secretion were measured using enzyme-linked immunosorbent assays. ABF® treatment downregulated lipid accumulation based on Oil Red O staining. ABF®-treated cells exhibited decreased mRNA and protein expression of LPL, C/EBPα, PPARγ, FABP2, and FAS. Moreover, ABF® treatment significantly increased adiponectin secretion and decreased leptin secretion. In conclusion, ABF® has anti-adipogenic effects on the differentiation of 3T3-L1 cells and may be used as an anti-obesity nutraceutical. [ABSTRACT FROM AUTHOR]

Agastache rugosa, or Korean mint, is an herb used as a spice, food additive and traditional medicinal ingredient. It has desirable effects, such as its antibacterial, antifungal and antioxidant properties. A. rugosa contains many phenolic compounds studied for their various health benefits, with the primary components being tilianin. A. rugosa extract (ARE), which was extracted with ethanol and freeze-dried, contained 21.14 ± 0.15 mg/g of tilianin with a total polyphenol content of 38.11 ± 0.88 mg/g. Next, the antiadipogenic effect of A. rugosa and tilianin was clarified using 3T3-L1 cells, which differentiate into adipocytes and develop lipid droplets. 3T3-L1 cells were treated with ARE or tilianin and lipid accumulation (%) was calculated through oil red O staining. Tilianin elicited dose-dependent decrease in lipid accumulation (% of positive control) (30 μM 92.10 ± 1.19%; 50 μM 69.25 ± 1.78%; 70 μM 54.86 ± 1.76%; non-differentiation 18.10 ± 0.32%), assessed by oil-red-O staining, whereas ARE treatments caused consistent diminution in lipid accumulation regardless of dose (100 μM 86.90 ± 4.97%; 200 μM 87.25 ± 4.34%; 400 μM 88.54 ± 2.27%; non-differentiation 17.96 ± 1.30%), indicating that both compounds have anti-obesity effects on adipocytes. Treatment with ARE lowered the mRNA (PPARγ; C/EBPα; FABP4; SREBP1; ACC; FAS) and protein (PPARγ; C/EBPα; SREBP1) levels of adipogenesis and lipogenesis-related factors. Tilianin showed a greater effect on the mRNA levels compared with ARE. Thus, tilianin and ARE may have anti-adipogenic and anti-lipogenic effects on 3T3-L1 cells and be possible candidates of obesity-related supplements. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the therapeutic efficacy and underlying molecular mechanisms of Bauhiniastatin-1 (BSTN1) to alleviate adiposity in diet-induced obese rodent model and in 3T3-L1 cells. Methods: BSTN1 was purified and confirmed through HPLC. In-vitro experiments such as MTT assay, Oil Red-O (ORO) stain, cellular lipid content, glycerol release and RT-PCR analysis were performed in 3T3-L1 cells in the presence and absence of BSTN1. In animal experiments, rats were divided into Group-I: normal pellet diet-fed, Group-II: HFD-fed, Groups-III, IV and V: HFD-fed BSTN1 (1.25, 2.5, and 5 mg/kg.b.wt./day/rat)-treated and Group-VI: HFD-fed Orlistat-treated. The rats were fed either normal diet or high fat diet (HFD) for 18 weeks and water ad-libitum. BSTN1 was orally administered from 13th week onwards to the selected HFD-fed groups. Body composition parameters, biochemical assays, histopathology examination and western blot analysis were performed to identify the predicted targets related to obesity. Molecular docking studies threw light on the binding interactions of BSTN1 against PPAR-γ, FAS and AMPK. Results: BSTN1 at 20 μM significantly (p < 0.001) inhibited adipocyte differentiation and lipid accumulation in 3T3-L1 cells. A conspicuous down-regulation in the mRNA expression levels of PPAR-γ, FAS and SREBP1 was observed but AMPK expression remained unchanged in BSTN1 treated 3T3-L1 cells. A substantial decrease in body weight gain, fat percent, total body fat, serum and liver lipid profile (except high-density lipoprotein), glucose, insulin and insulin resistance in BSTN1 treated rats was noticed in a dose dependent manner. In BSTN1 (5 mg/kg.b.wt.)-treated groups significantly (p < 0.01) elevated plasma adiponectin level but reduced leptin level as well as fall in serum AST and ALT were noticed. Further, the disturbed structural integrity and architecture of adipose and hepatic tissues due to high fat diet feeding were considerably recovered with BSTN1 treatment. Down-regulation in the protein expression level of PPAR-γ and activation of AMPK through phosphorylation was observed in BSTN1 treated rats than the untreated. Molecular docking studies revealed strong binding interactions of BSTN1 against PPAR-γ and AMPK and thus supported the experimental results. Conclusion: Taken together, the results suggest that BSTN1 could be a promising pharmacological molecule in the treatment of obesity and dyslipidemia. [ABSTRACT FROM AUTHOR]

Obesity is a complex disease spreading in the world. In our previous studies, chlorogenic acid (CGA) and caffeine had ever been reported to reduce the body weight gain and fat accumulation in mice. This study investigated the anti‐obesity effect of CGA and caffeine on 3T3‐L1 cells. According to triglyceride (TG) assay and Oil‐Red O staining, 40 μg/ml CGA and 160 μg/ml caffeine reduced TG content. Moreover, CGA + caffeine inhibited the mRNA expression of major adipogenic markers, PPAR‐γ2, and C/EBPα in the metaphase and anaphase stages of differentiation induction (Day 2 and 4). CGA + caffeine improved P‐AMPK/AMPK accompanied by decreasing the expression of GPDH and FAS to depress the lipid synthesis, increasing the mRNA expression of ACO and CAT to promote fatty acid oxidation and up‐regulated the expression of hydrolysis‐related enzyme adipose TG lipase (ATGL) and P‐HSL/HSL. Furthermore, CGA + caffeine improved the expression of Glut4 which promoted the glucose transport. Taken together, these data demonstrated CGA + caffeine inhibited 3T3‐L1 cells differentiation in the middle and late stages and reduced the fat accumulation through AMPK pathway by regulating the fat metabolism‐related enzyme in 3T3‐L1 cells to attenuates adipogenesis. Practical applications: The aim of this study was to elucidate the potential role of chlorogenic acid and caffeine in the treatment of obesity. [ABSTRACT FROM AUTHOR]

Liver fibrosis, with the characterization of progressive accumulation of extracellular matrix (ECM), is the common pathologic feature in the process of chronic liver disease. Hepatic stellate cells (HSCs) which are activated and differentiate into proliferative and contractile myofibroblasts are recognized as the main drivers of fibrosis. Obesity-related adipocytokine dysregulation is known to accelerate liver fibrosis progression, but the direct fibrogenic effect of mature adipocytes on HSCs has been rarely reported. Therefore, the purpose of this study was to explore the fibrogenic effect of adipocyte 3T3-L1 cells on hepatic stellate LX-2 cells. The results showed that incubating LX-2 cells with the supernatant of 3T3-L1 adipocytes triggered the expression of ECM related proteins, such as α-smooth muscle actin (α-SMA), type I collagen (CO-I), and activated TGF β/Smad2/3 signaling pathway in LX-2 cells. In addition, 3T3-L1 cells inhibited insulin sensitivity, activated endoplasmic reticulum stress and autophagy to promote the development of fibrosis. These results supported the notion that mature adipocytes can directly activate hepatic stellate cells, and the establishment of an in vitro model of adipocytes on HSCs provides an insight into screening of drugs for liver diseases, such as nonalcoholic fatty liver disease. [ABSTRACT FROM AUTHOR]

4-Thiazolidinones and related derivatives are regarded as privileged structures in medicinal chemistry and a source of new drug-like compounds. To date it is known that thiazolidinones are able to induce CYP1A1 activity in 3T3-L1 cells. Therefore, to extend the knowledge of the mechanism of thiazolidinones in the cell, four chemically synthesized heterocycles were tested on 3T3-L1 cells. The 3T3-L1 cells were exposed to Les-2194, Les-3640, Les-5935, and Les-6166. Our study showed that 1 μM βNF, Les-2194, and Les-6166 decreased the expression of Ahr mRNA. In turn, βNF, Les-2194, and Les-3640 increased the Cyp1a1 mRNA expression at the same time interval. On the other hand, Les-5935 was found to decrease the Cyp1a1 mRNA expression. Interestingly, the expression of Cyp1a2 mRNA was activated only by βNF and Les-2194. The expression of Cyp1b1 mRNA in the 3T3 cell line increased after the βNF and Les-2194 treatment but declined after the exposure to Les-5935 and Les-6166. Moreover, the Les-2194 and Les-5935 compounds were shown to increase the activity of EROD, MROD, and PROD. Les-3640 increased the activity of EROD and decreased the activity of PROD. In turn, the treatment with Les-6166 resulted in an increase in the activity of EROD and a decrease in the activity of MROD and PROD in the 3T3-L1 cells. [ABSTRACT FROM AUTHOR]

Amomum tsao-ko (Zingiberaceae), an important traditional medicinal herb, possesses many biological activities, including anti-inflammatory effects. Though the anti-obesity properties of the crude ethanol extract of A. tsao-ko fruits have been reported, the anti-adipogenic properties of its phytochemical constituents have not been reported. Therefore, in the present study, we isolated the active constituents of A. tsao-ko and investigated their anti-adipogenic effects. The bioassay-guided isolation of the phytochemicals from the ethanol extract of A. tsao-ko fruits identified four bioactive compounds, comprising one fatty acid (1), one sesquiterpene alcohol (2), and two phenolic compounds (3 and 4). Their structures were elucidated by a combination of 1D and/or 2D nuclear magnetic resonance and mass spectrometry. The anti-adipogenic activities of the four compounds evaluated by Oil Red O staining in 3T3-L1 cells revealed that the treatment with the isolated compounds 1 and 3 reduced the lipid accumulation in 3T3-L1 adipocytes more strongly than the compounds 2 and 4, in a dose-dependent manner. [ABSTRACT FROM AUTHOR]