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Green tea possesses a range of beneficial effects, including anti-obesity, antioxidant, and anti-inflammatory properties, owing to its biologically active components, primarily catechins such as epicatechin (EC), epicatechin gallate (ECG), epigallocatechin (EGC), and epigallocatechin gallate (EGCG). However, few studies have investigated the four catechin monomers simultaneously, and the molecular mechanisms of their anti-obesity effects have not been fully elucidated. In this study, we investigated the effects of four catechin monomers on the differentiation of 3T3-L1 preadipocytes of mice. Our findings demonstrated that four catechin monomers EC/ECG/EGC/EGCG (12, 25, 50 µM) dose-dependently inhibited the differentiation of 3T3-L1 preadipocytes and reduced triglyceride content. EGCG exhibited the most potent inhibitory effect with an optimal concentration of 50 µM. In addition, transcriptome sequencing and lipidomic analysis of EGCG-treated 3T3-L1 preadipocytes revealed that Ptgs2 and Pim1 were the most differentially expressed genes involved in regulating adipocyte differentiation. The results suggested that EGCG up-regulated the expression of the Pla2g2e gene and down-regulated the expression of the Pla2g4a and Pla2g2a genes via the glycerophospholipid metabolic pathway, which subsequently elevated lysophosphatidylcholine (LPC) levels, influencing the differentiation process of 3T3-L1 preadipocytes. [ABSTRACT FROM AUTHOR]

Objective: To study the effect and mechanism of luteoloside on the differentiation and lipid metabolism of 3T3-L1 preadipocytes. Methods: The cytotoxicity of different concentrations of luteoloside on 3T3-L1 preadipocyte was detected using methyl thiazolyl tetrazolium (MTT) assay. Cell differentiation was induced by the cocktail method, and lipid droplets were observed by oil red O staining. The contents of triglyceride and total cholesterol in differentiated cells, as well as the secretion of related inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-10, leptin (LEP) and Adiponectin (ADPN) were measured. The relative expression levels of peroxisome proliferators activated receptor (PPAR) γ, CCAAT-enhancer-binding proteins (C/EBP)-α, sterol regulatory element-binding protein 1 (SREBP1), PPARα, uncoupling protein 1 (UCP-1), and carnitine palmitoyltransferase 1 (CPT-1) were analyzed by Western blotting. Results: The survival rate of 3T3-L1 preadipocytes decreased with the increase in luteoloside concentration, being above 80% at luteoloside concentrations of 5–60 μg/mL. Treatment with luteoloside significantly inhibited the differentiation of 3T3-L1 preadipocytes and reduced lipid accumulation. Luteoloside down-regulated the contents of TC and TG, the secretion of TNF-α, IL-6 and LEP, but up-regulated the secretion of IL-10 and ADPN. Luteoloside down-regulated the protein expression levels of PPARγ, C/EBP-α and SREBP1, but up-regulated those of PPARα, UCP-1 and CPT-1. Conclusion: Luteoloside can suppression the differentiation and lipid metabolism of 3T3-L1 preadipocytes. The mechanism may be that luteoloside down-regulates the protein expression of PPARγ, C/EBP-α, and SREBP1 to inhibit fat synthesis. Furthermore, it activates PPARα to upregulate downstream proteins and promote fat consumption. Luteoloside may also regulate the secretion of cytokines and promote cellular lipolysis, thus improving lipid metabolism imbalance.

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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.)

Obesity is one of the more complicated diseases, it can induce numerous life‐threatening diseases mainly diabetes mellitus, cardiovascular disease, hypertension, and certain cancers. In this study, we assessed the efficacy of bacoside‐A (a dammarane‐type triterpenoid saponin derived from the plant Bacopa monniera Linn.) on the adipogenesis of 3T3‐L1 preadipocytes. Results of this study illustrated that bacoside‐A decreased the differentiation of 3T3‐L1 cell, as evidenced by diminution of lipid droplets, which contains triglycerides and other lipids. During the differentiation process, transcription factors, which are mainly participating in adipogenesis such us CCAAT/enhancer‐binding protein α (C/EBPα) and C/EBPβ, peroxisome proliferator‐activated receptor‐γ (PPARγ), and sterol regulatory element‐binding protein‐1c (SREBP‐1c), expressions were significantly suppressed by bacoside‐A. In addition, bacoside‐A showed a potent reduction in genes precise to adipocytes such as lipoprotein lipase (LPL), fatty acid synthase (FAS), adipocyte fatty acid‐binding protein (FABP4), and leptin expressions. Further, bacoside‐A stimulated the phosphorylation of acetyl CoA carboxylase (ACC) and AMP‐activated protein kinase (AMPK). These results demonstrated that bacoside‐A has anti‐adipogenic effects by regulating the transcription factors involved in adipocyte differentiation. Therefore, bacoside‐A might be considered as a potent therapeutic agent for alleviating obesity and hyperlipidemia. [ABSTRACT FROM AUTHOR]

Copyright of Food & Machinery is the property of Food & Machinery Editorial Office 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.)

[Objective] β-Sitosterol-modified ginsenoside Rb1 liposomes （β-Rb1-Lip） were prepared to reduce the degradation of Rb1 and enhance the lipid-lowering effects of ginsenoside Rb1. [Methods] β-sitosterol-modified ginsenoside Rb1 liposomes were prepared by the thin-film hydration method. The biosafety of the liposomes was assessed using the MTT assay. The inhibition of lipid droplet accumulation in 3T3-L1 adipocytes by β-Rb1-Lip was investigated using Oil Red O staining and triglyceride （TG） content measurement with an enzyme-linked immunosorbent assay. [Results] The prepared β-Rb1-Lip liposomes had an encapsulation efficiency of 83.74% and an average particle size of 198 nm. The release rate of Rb1 from β-Rb1-Lip was about 80% within 12 hours, demonstrating a good sustained-release effect. Regarding lipid-lowering activity, β-Rb1-Lip at 50 μmol/L showed a significant inhibitory rate of intracellular lipid droplets. Compared to the same concentration of Rb1 monomer, β-Rb1-Lip had a more significant inhibitory effect on the accumulation of lipid droplets in 3T3-L1 cells and did not exhibit cytotoxicity. [Conclusion] β-Rb1-Lip has high encapsulation efficiency, small particle size, and obvious sustained-release characteristics. It can continuously release the active component ginsenoside Rb1, enhance its lipid-lowering efficacy, and reduce the dosage, providing support for the development of Rb1 lipid-lowering products.

Background. Obesity and related metabolic diseases are becoming a worldwide epidemic, leading to increased mortality and heavy medical costs. Our Chinese herbal formula Xiao-Gao-Jiang-Zhuo (XGJZ) has remarkable effects on curing obese patients in the clinic, but the cellular and molecular basis remains unknown. This study aimed to reveal the molecular mechanism involved in adipogenesis in vitro. Methods. Chinese herbal formula XGJZ-containing serum was prepared from XGJZ-treated obesity model rats. The function of XGJZ-containing serum was validated in 3T3-L1 preadipocytes. Oil O staining was performed to determine intracellular lipid accumulation in differentiated 3T3-L1 cells. The expression of pro-adipogenic transcription factors was measured to further validate the adipogenesis of 3T3-L1 adipocytes. The contents of triglyceride (TG), free fatty acid (FFA), and glycerin, along with the activities of lipid metabolism-related enzymes (including FAT, FATP1, DGAT, GPAT, ATGL, and HSL) were measured to study the lipogenesis in 3T3-L1 adipocytes. Results. XGJZ-containing serum inhibited 3T3-L1 differentiation, decreased intracellular lipid accumulation, and suppressed the expression of pro-adipogenic transcription factors in differentiated 3T3-L1 cells. The contents of TG, FFA, and glycerin were decreased when treated with XGJZ-containing serum, which also modulated lipid metabolism-related enzyme activities. The activities of fatty acid transporters (FAT, FATP1) and lipid mobilization enzymes (ATGL, HSL) were promoted, while activities of triglyceride biosynthesis enzymes (DGAT, GPAT) were attenuated in differentiated 3T3-L1 cells. Conclusion. XGJZ-containing serum has inhibitory effects on adipogenesis in 3T3-L1 preadipocytes, affirming the effect of XGJZ in treating obesity. It provides evidence for the mechanism of obesity.

Tetrabromobisphenol A (TBBPA), a widely-used brominated flame retardant, has been revealed to exert endocrine disrupting effects and induce adipogenesis. Given the high structural similarities of TBBPA analogues and their increasing exposure risks, their effects on lipid metabolism are necessary to be explored. Herein, 9 representative TBBPA analogues were screened for their interference on 3T3-L1 preadipocyte adipogenesis, differentiation of C3H10T1/2 mesenchymal stem cells (MSCs) to brown adipocytes, and lipid accumulation of HepG2 cells. TBBPA bis(2-hydroxyethyl ether) (TBBPA-BHEE), TBBPA mono(2-hydroxyethyl ether) (TBBPA-MHEE), TBBPA bis(glycidyl ether) (TBBPA-BGE), and TBBPA mono(glycidyl ether) (TBBPA-MGE) were found to induce adipogenesis in 3T3-L1 preadipocytes to different extends, as evidenced by the upregulated intracellular lipid generation and expressions of adipogenesis-related biomarkers. TBBPA-BHEE exhibited a stronger obesogenic effect than did TBBPA. In contrast, the test chemicals had a weak impact on the differentiation process of C3H10T1/2 MSCs to brown adipocytes. As for hepatic lipid formation test, only TBBPA mono(allyl ether) (TBBPA-MAE) was found to significantly promote triglyceride (TG) accumulation in HepG2 cells, and the effective exposure concentration of the chemical under oleic acid (OA) co-exposure was lower than that without OA co-exposure. Collectively, TBBPA analogues may perturb lipid metabolism in multiple tissues, which varies with the test tissues. The findings highlight the potential health risks of this kind of emerging chemicals in inducing obesity, non-alcoholic fatty liver disease (NAFLD) and other lipid metabolism disorders, especially under the conditions in conjunction with high-fat diets.

Boesenbergia rotunda has been used as an antiobesity agent by suppressing adipogenesis. This study aimed to investigate the biological activity of B. rotunda on preadipocyte cells and to evaluate the effectiveness and safety of using B. rotunda extract in a capsaicin-loaded body-firming formulation. The antiadipogenesis of B. rotunda ethanolic extract was evaluated in 3T3-L1 preadipocyte cells. After the application of the B. rotunda extract-loaded body-firming formulation on the skin of volunteers for 28 d, thigh circumference, melanin index, and skin erythema were investigated. The results showed that the ethanolic extract of B. rotunda was not toxic toward 3T3-L1 cells at concentrations lower than 20 µg/mL, with antiadipogenesis of the B. rotunda extract occurring at a concentration of 1 µg/mL. The B. rotunda extract containing panduratin A was mixed with capsaicin body-firming products and successfully permeated into and through the skin. Applying this formulation to the thighs of the volunteers two times a day for 21 days led to a significant reduction in thigh circumference and melanin index. A slight elevation in skin erythema was observed, but there was no significant increase in redness or pain. In conclusion, the B. rotunda extract contained bioactive compounds that inhibited antiadipogenesis. The formulations containing B. rotunda extract and capsaicin showed potential as effective body-firming products. [ABSTRACT FROM AUTHOR]

The augmentation of adipocytes in the adipose tissues brings disordered pathophysiological conditions, including type 2 diabetes, hyperlipidemia, hypertension, cardiovascular disease, and cancer. The phenolic antioxidant 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) prevents oxidative stress as radical scavenging in cells. However, the role of the disorder as a pharmacologic factor has been poorly understood. This study elucidates the regulatory effects of DHMBA on adipogenesis in mouse 3T3-L1 adipocytes in vitro. The 3T3-L1 preadipocytes were cultured in DMEM containing 10% calf fetal serum in the presence of DHMBA. Culturing with DHMBA repressed the growth of 3T3-L1 preadipocytes cultured in a medium without differentiation factors. Interestingly, when 3T3-L1 preadipocytes were cultured in a medium including differentiation factors containing insulin, DHMBA did not affect the number of cells with the differentiation process of adipogenesis. Culturing with DHMBA (1, 10, or 100 μM) inhibited lipid accumulation in adipocytes and repressed adipogenesis in 3T3-L1 cells. The potent inhibitory effects of DHMBA on adipogenesis were seen at the later stage of culture. Adipogenesis was inhibited by the presence of wortmannin, PD98059, or Bay 11-7082, which are inhibitors of pathways related to insulin signaling pathway. Notably, the suppressive effects of DHMBA on adipogenesis were expressed by the presence of these inhibitors. DHMBA treatment declined the levels of PPARy and C/EBPα related to preadipocyte differentiation and PI3 kinase 100α, Akt, MAPK, phosphor-MAPK, and mTOR implicated in the insulin signaling pathway, leading to adipogenesis promotion. Thus, DHMBA may inhibit adipogenesis via regulating diverse signaling pathways, providing a new strategy for the therapy of obesity.

Aim: The present study is aimed to examine olvanil's effect in preadipocyte cell culture and on a murine model of diet-induced obesity. We hypothesized that olvanil by being a capsaicinoid will reduce the differentiation to mature adipocytes and reduce the weight of fat tissue in the studied mice. Methods: To determine the effect of olvanil on adipogenesis, 3T3-L1 cells were cultured. Oil red staining was performed to determine lipid accumulation, whereas triglycerides were measured by biochemical determination. Expression of PPAR-γ and PREF-1 were measured by RT-PCR. Therefore, male C57BL/6J mice (CICUAL:2018-020B) were fed with a high-fat diet for 12 weeks to develop a murine model of diet-induced obesity (DIO). Animals were separated into 6 experimental groups: control (standard diet), DIO (high-fat diet), DIO + orlistat (10 mg/kg), DIO + olvanil (10 mg/kg), DIO + olvanil (25 mg/kg) and DIO + olvanil (50 mg/kg), olvanil was administered for 4 weeks. Results: Olvanil inhibits adipogenesis, reduces lipid accumulation and triglycerides in 3T3-L1 adipocytes. PPAR-ɣ gene expression was suppressed while PREF-1 was increased in adipocytes treated with olvanil. Whereas protein expression of FABP4 and PPAR-ɣ decreases significantly with olvanil. The results suggest that olvanil can inhibit the differentiation of preadipocytes to adipocytes through the overexpression or maintenance of PREF-1 levels and the suppression of PPAR-ɣ, and FABP4. Therefore, in diet-induced obesity in mice, olvanil decreases fat accumulation in the body and improve lipid profile by decreasing LDL, VLDL and triglycerides in serum. Conclusion: Olvanil inhibits adipocyte differentiation in 3T3-L1 cells and reduces fat accumulation and ameliorate lipid profile in diet-induced obese mice.

The augmentation of adipocytes in the adipose tissues brings disordered pathophysiological conditions, including type 2 diabetes, hyperlipidemia, hypertension, cardiovascular disease, and cancer. The phenolic antioxidant 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) prevents oxidative stress as radical scavenging in cells. However, the role of the disorder as a pharmacologic factor has been poorly understood. This study elucidates the regulatory effects of DHMBA on adipogenesis in mouse 3T3-L1 adipocytes in vitro. The 3T3-L1 preadipocytes were cultured in DMEM containing 10% calf fetal serum in the presence of DHMBA. Culturing with DHMBA repressed the growth of 3T3-L1 preadipocytes cultured in a medium without differentiation factors. Interestingly, when 3T3-L1 preadipocytes were cultured in a medium including differentiation factors containing insulin, DHMBA did not affect the number of cells with the differentiation process of adipogenesis. Culturing with DHMBA (1, 10, or 100 μM) inhibited lipid accumulation in adipocytes and repressed adipogenesis in 3T3-L1 cells. The potent inhibitory effects of DHMBA on adipogenesis were seen at the later stage of culture. Adipogenesis was inhibited by the presence of wortmannin, PD98059, or Bay 11-7082, which are inhibitors of pathways related to insulin signaling pathway. Notably, the suppressive effects of DHMBA on adipogenesis were expressed by the presence of these inhibitors. DHMBA treatment declined the levels of PPARy and C/EBPα related to preadipocyte differentiation and PI3 kinase 100α, Akt, MAPK, phosphor-MAPK, and mTOR implicated in the insulin signaling pathway, leading to adipogenesis promotion. Thus, DHMBA may inhibit adipogenesis via regulating diverse signaling pathways, providing a new strategy for the therapy of obesity. [ABSTRACT FROM AUTHOR]

Chlorogenic acid has been widely reported to profoundly influence preadipocyte differentiation but without consensus being achieved. In the current study, 3 T3-L1 preadipocytes were differentiated using adipogenic cocktail with or without chlorogenic acid. The influences of chlorogenic acid on the mRNA, lipid and phospho-kinase profiles were evaluated using transcriptomic analysis, lipidomic analysis and phospho-kinase array assay. The obtained results showed that chlorogenic acid treatment during 3T3-L1 cells differentiation altered a series of genes transcription, resulted in obvious changes in the contents of various lipids, and inhibited phosphorylation of several adipogenesis-related kinases. Overall, the observed changes in transcriptomic, lipidomic and phospho-kinase profiles in the current study favor the findings that chlorogenic acid may inhibit the differentiation of 3T3-L1 preadipocytes. In conclusion, the findings from this study highlighted the therapeutic potential of chlorogenic acid for the treatment of obesity.

Adipose tissues are closely related to physiological functions and pathological conditions in most organs. Although differentiated 3T3-L1 preadipocytes have been used for in vitro adipose studies, the difference in cellular characteristics of adipogenic differentiation in two-dimensional (2D) culture and three-dimensional (3D) culture remain unclear. In this study, we evaluated gene expression patterns using RNA sequencing and metabolic functions using an extracellular flux analyzer in 3T3-L1 preadipocytes with and without adipogenic induction in 2D culture and 3D culture. In 2D culture, 565 up-regulated genes and 391 down-regulated genes were identified as differentially expressed genes (DEGs) by adipogenic induction of 3T3-L1 preadipocytes, whereas only 69 up-regulated genes and 59 down-regulated genes were identified as DEGs in 3D culture. Ingenuity Pathway Analysis (IPA) revealed that genes associated with lipid metabolism were identified as 2 out of the top 3 causal networks related to diseases and function in 3D spheroids, whereas only one network related to lipid metabolism was identified within the top 9 of these causal networks in the 2D planar cells, suggesting that adipogenic induction in the 3D culture condition exhibits a more adipocyte-specific gene expression pattern in 3T3-L1 preadipocytes. Real-time metabolic analysis revealed that the metabolic capacity shifted from glycolysis to mitochondrial respiration in differentiated 3T3-L1 cells in the 3D culture condition but not in those in the 2D cultured condition, suggesting that adipogenic differentiation in 3D culture induces a metabolic phenotype of well-differentiated adipocytes. Consistently, expression levels of mitochondria-encoded genes including mt-Nd6, mt-Cytb, and mt-Co1 were significantly increased by adipogenic induction of 3T3-L1 preadipocytes in 3D culture compared with those in 2D culture. Taken together, the findings suggest that induction of adipogenesis in 3D culture provides a more adipocyte-specific gene expression pattern and enhances mitochondrial respiration, resulting in more adipocyte-like cellular properties.

During the worldwide COVID-19 outbreak, there was an increase in the prevalence of obesity, including childhood obesity, due to which the awareness of obesity and interest in treatment increased. Accordingly, we describe EJF (Euscaphis japonica Kanitz fruit) extract as a candidate for naturally derived antiobesity agents. In this study, we found that EJF is involved in the early stage of adipogenic differentiation in vitro and finally inhibits adipogenesis. We propose two mechanisms for the antiobesity effect of EJF. First, EJF inhibits MDI-induced mitotic clonal expansion (MCE) by inducing cell cycle arrest at the initiation of adipogenic differentiation. The second aims to regulate stability and activation at the protein level of IRS1, which initiates differentiation in the early stage of differentiation. As a result, it was found that the activation of Akt decreased, leading to the inhibition of the expression of adipogenesis-related transcription factors (PPARγ, C/EBPα) and the subsequent suppression of adipogenic differentiation. In summary, we suggest that EJF can inhibit adipogenesis and lipid accumulation by suppressing the early stage of adipogenic differentiation in 3T3-L1 adipocytes. These findings indicate that EJF's functionality could be beneficial in the treatment of obesity, particularly childhood obesity associated with adipocyte hyperplasia. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effect of Grape Seed Proanthocyanidin (GSP) on fat metabolism and adipocytokines in obese rats. Fifty 5-week-old rats were randomly assigned to five groups (n = 10 per group) and given either a basal diet, a high-fat diet, or a high-fat diet supplemented with GSP (25, 50, and 100 mg/d) per group. The experiment lasted for five weeks, including a one-week adaptation period and a four-week treatment period. At the end of the experimental period, serum and adipose tissue samples were collected and analyzed. Additionally, we co-cultured 3T3-L1 preadipocytes with varying concentrations of GSP to explore its effect on adipocyte metabolism. The results demonstrated that GSP supplementation reduced weight, daily gain, and abdominal fat weight coefficient (p < 0.05). It also decreased levels of glucose, cholesterol (TC) (p < 0.05), triglycerides (TG) (p < 0.05), low-density lipoprotein (LDL), cyclooxygenase-2 (COX-2), and interleukin-6 (IL-6) in adipose tissue. Furthermore, GSP addition caused adipocyte crumpling in vitro and reduced the mRNA expression of COX-2, LEP, and TNF-α in adipocytes in vitro. These findings provide compelling evidence for exploring the role of GSP in the prevention and treatment of obesity and related diseases. [ABSTRACT FROM AUTHOR]

Lanthanum (La) as a rare earth element is widely used in agriculture, industry, and medicine. It has been suggested in several studies that La might influence glycolipid metabolism in vivo. In this study, we used 3T3‐L1 preadipocytes as in vitro cell model to elucidate the effects of La(NO3)3 on adipogenesis and the underlying mechanisms. The results showed that La(NO3)3 could inhibit the adipogenic differentiation of 3T3‐L1 preadipocytes, which showed a decrease in lipid accumulation and the downregulation of specific adipogenic transcription factors. La(NO3)3 exerted its inhibitory effect mainly at the early differentiation stage. Furthermore, La(NO3)3 influenced the S‐phase entry and cell cycle process during the mitotic clonal expansion and regulated the phosphorylation of signal transducer and activator of transcription 3 (STAT3) and expressions of the proteins in phosphatidylinositol 3‐kinase (PI3K)/Akt pathway at the early stage of differentiation. Besides, La(NO3)3 upregulated the expressions of wnt10b mRNA and β‐catenin protein and promoted the nucleus translocation of β‐catenin. Additionally, we found that La(NO3)3 could promote the growth of 3T3‐L1 preadipocytes both with and without MDI (3‐isobutyl‐1‐methylxanthine [IBMX], dexamethasone [Dex], and insulin) stimulation. Collectively, these results indicated that La(NO3)3 could inhibit adipogenesis in 3T3‐L1 preadipocytes and influence cell proliferation. Lanthanum (La) as a rare earth element is widely used in many aspects and might influence glycolipid metabolism in vivo. In this study, we used 3T3‐L1 preadipocytes as in vitro cell model to elucidate the effects of La(NO3)3 on adipogenesis. The results showed that La(NO3)3 could inhibit the adipogenic differentiation of 3T3‐L1 preadipocytes and influence the cell cycle process during the mitotic clonal expansion. Also, the wnt/β‐catenin signaling pathway might be involved in the inhibitory effect of La(NO3)3. [ABSTRACT FROM AUTHOR]

Boesenbergia rotunda has been used as an antiobesity agent by suppressing adipogenesis. This study aimed to investigate the biological activity of B. rotunda on preadipocyte cells and to evaluate the effectiveness and safety of using B. rotunda extract in a capsaicin-loaded body-firming formulation. The antiadipogenesis of B. rotunda ethanolic extract was evaluated in 3T3-L1 preadipocyte cells. After the application of the B. rotunda extract-loaded body-firming formulation on the skin of volunteers for 28 d, thigh circumference, melanin index, and skin erythema were investigated. The results showed that the ethanolic extract of B. rotunda was not toxic toward 3T3-L1 cells at concentrations lower than 20 µg/mL, with antiadipogenesis of the B. rotunda extract occurring at a concentration of 1 µg/mL. The B. rotunda extract containing panduratin A was mixed with capsaicin body-firming products and successfully permeated into and through the skin. Applying this formulation to the thighs of the volunteers two times a day for 21 days led to a significant reduction in thigh circumference and melanin index. A slight elevation in skin erythema was observed, but there was no significant increase in redness or pain. In conclusion, the B. rotunda extract contained bioactive compounds that inhibited antiadipogenesis. The formulations containing B. rotunda extract and capsaicin showed potential as effective body-firming products.

Background: Withania somnifera (Solanaceae), generally known as Indian ginseng, is a medicinal plant that is used in Ayurvedic practice for promoting health and longevity. This study aims to identify the bioactive metabolites from Indian ginseng and elucidate their structures. Methods: Withanolides were purified by chromatographic techniques, including HPLC coupled with LC/MS. Chemical structures of isolated withanolides were clarified by analyzing the spectroscopic data from 1D and 2D NMR, and HR-ESIMS experiment. Absolute configurations of the withanolides were established by the application of NMR chemical shifts and ECD calculations. Anti-adipogenic activities of isolates were evaluated using 3T3-L1 preadipocytes with Oil Red O staining and quantitative real-time PCR (qPCR). Results: Phytochemical examination of the roots of Indian ginseng afforded to the isolation of six withanolides (1–6), including three novel withanolides, withasilolides G–I (1–3). All the six compounds inhibited adipogenesis and suppressed the enlargement of lipid droplets, compared to those of the control. Additionally, the mRNA expression levels of Fabp4 and Adipsin, the adipocyte markers decreased noticeably following treatment with 25 μM of 1–6. The active compounds (1–6) also promoted lipid metabolism by upregulating the expression of the lipolytic genes HSL and ATGL and downregulating the expression of the lipogenic gene SREBP1. Conclusion: The results of our experimental studies suggest that the withasilolides identified herein have anti-adipogenic potential and can be considered for the development of therapeutic strategies against adipogenesis in obesity. Our study also provides a mechanistic rationale for using Indian ginseng as a potential therapeutic agent against obesity and related metabolic diseases.

As a major cause of mortality, cardiovascular disease is associated with obesity and diabetes. However, the molecular mechanism by which diabetes-obesity causes cardiovascular complications is largely unknown. In this study, the crosstalk mediated by 3T3-L1 preadipocytes and mouse retina microvascular endothelial cells (mRMECs) was determined after co-culturing performed with a Transwell system or measuring exosome uptake by mRMECs. CCK-8 assays, EdU incorporation assays, TUNEL staining, and ELISAs were used to evaluate the functions of mRMECs. Related protein markers were analyzed by western blotting. Our results showed that LINC00968 levels were significantly elevated in the exosomes derived from H-Glu-induced 3T3-L1 preadipocytes. Both H-Glu treatment and co-culture with 3T3-L1 cells damaged mRMECs, as indicated by lower rates of proliferation and higher rates of apoptosis and cell adhesion molecule expression, as well as by induced inflammation and oxidative stress, which were enhanced by combined H-Glu and co-culture treatment. Furthermore, H-Glu and co-culture treatment increased LINC00968 expression in mRMECs, and the exosomes collected from 3T3-L1 cells had a similar effect. Functionally, LINC00968 inhibition protected mRMECs against the effects of H-Glu and co-culture treatment, while LINC00968 played the opposite role. LINC00968 was found to target miR-361–5p, and TRAF3 was identified as a target gene of miR-361–5p. Finally, miR-361–5p overexpression alleviated the effects of LINC00968 on H-Glu-induced mRMEC dysfunction in vitro. In conclusion, our results indicated that in an H-glu environment, adipocyte exosomes damage microvascular endothelial cells via a LINC00968/miR-361–5p/TRAF3 signaling pathway, which could possibly serve as a target for treating diabetes-obesity-triggered microvascular complications. [ABSTRACT FROM AUTHOR]

Corn peptide (CP) is a small, natural, biologically active peptide obtained by protease-catalysed hydrolysis of corn. CP exerts antihypertensive, hypoglycaemic, antihyperlipidemic, antioxidant, and antitumor effects, as well as prevents cardiovascular and cerebrovascular diseases. Although CP plays a role in preventing obesity-related diseases, its role in reducing obesity has not yet been determined. In this study, we analysed the inhibitory effects of CP on lipid droplet accumulation in 3T3-L1 preadipocytes and high-fat diet (HFD)-induced C57BL/6J Obese Mice. The results show that CP could inhibit preadipocyte differentiation and oil accumulation in 3T3-L1 preadipocytes. Oral CP administration reduced serum triglyceride (TG) content, epididymal fat weight, abnormal liver fat droplet accumulation, and C/EBPα expression. Furthermore, combination of CP administration and exercise reduced body, liver, and adipose tissue weights; decreased serum total cholesterol (TC), triglyceride and low-density lipoprotein (LDL) levels; and inhibited hepatic lipid droplet accumulations and epididymal fat cell hypertrophy. Additionally, this combination inhibited the expression of transcription factors, C/EBPα, C/EBPβ, and PPARγ, and adipogenic factors, FABP4 in mice. In conclusion, oral administration of CP inhibited lipid droplet accumulation and counteracted HFD-induced obesity in mice. [ABSTRACT FROM AUTHOR]

Maternal body mass index is associated with breast milk (BM) fatty acid composition. This study investigated the effects of BM omega (n)-6:n-3 polyunsaturated fatty acids (PUFAs) from non-obese women and women with obesity on the process of adipogenesis in 3T3-L1 preadipocytes. BM samples were collected from non-obese women (BMNO) and women with obesity (BMO) at one month postpartum. The fatty acid composition was measured, and BMNO and BMO groups with the lowest (Q1) and highest (Q4) quartiles of n-6:n-3 PUFA ratios were identified. 3T3-L1 preadipocytes were differentiated in the presence or absence of BM. Lipid accumulation and the expression of genes involved in lipogenesis and lipolysis were measured. Treatment with BMNO containing high (vs. low) n-6:n-3 PUFA ratios significantly increased the mRNA expression of lipogenic genes (acetyl-CoA carboxylase, fatty acid synthase, and stearoyl-CoA desaturase); however, there was no effect when cells were treated with BMO (with either low or high n-6:n-3 PUFA ratios). Treatment with BMO (high n-6:n-3 PUFA ratio) caused larger lipid droplets. Our findings demonstrated that BMNO with a high n-6:n-3 PUFA ratio was associated with a higher expression of lipogenic genes, while BMO with a high n-6:n-3 PUFA ratio showed larger lipid droplets, suggesting adipocyte dysfunction. These findings may have implications in the BM-mediated programming of childhood obesity. [ABSTRACT FROM AUTHOR]

Obesogens are a subset of endocrine disruptor chemicals (EDCs) that cause obesity. The typical EDC 4‐nonylphenol (4‐NP) has been identified as an obesogen. However, the in vitro effects of 4‐NP on adipogenesis remain unclear. In this study, 3T3‐L1 preadipocytes and C3H/10T1/2 mesenchymal stem cells (MSCs) were used to investigate the influence of 4‐NP on adipogenesis. The differentiation protocols for 3T3‐L1 preadipocytes and C3H/10T1/2 MSCs took 8 and 12 days, respectively, beginning at Day 0. In differentiated 3T3‐L1 preadipocytes, 20 μM 4‐NP decreased cell viability on Days 4 and 8. Exposure to 4‐NP inhibited triglyceride (TG) accumulation and adipogenic marker expression on Days 0–8, but the inhibitory effects were weaker on Days 2–8. The protein expression of pSTAT3 or STAT3 decreased on Days 0–8 and 2–8. Conversely, 4‐NP promoted TG accumulation and the adipogenic marker expression in C3H/10T1/2 adipocytes. The opposing effects were attributed to physiological differences between the two cell lines. The 3T3‐L1 preadipocytes are dependent on mitotic clonal expansion (MCE) to drive differentiation, while C3H/10T1/2MSCs and human preadipocytes are not. Additionally, 4‐NP downregulated β‐catenin expression in C3H/10T1/2 adipocytes. Accordingly, we hypothesized that 4‐NP promotes adipogenesis. The role of the canonical Wnt pathway in the promotion of adipogenesis by 4‐NP requires further validation. This study provides new insights into the mechanisms and appropriate risk management of 4‐NP. This study aimed to investigate the effects of 4‐nonylphenol (4‐NP) on adipogenesis in vitro. The results showed that 4‐NP inhibited 3T3‐L1 preadipocyte differentiation but promoted C3H/10T1/2 mesenchymal stem cell (MSC) adipogenesis. The opposing effects were likely due to physiological differences between the two cell lines. The 3T3‐L1 preadipocytes are dependent on MCE to drive differentiation, whereas C3H/10T1/2 MSCs are not. We found that 4‐NP promoted adipogenesis, and new insights into the mechanisms have been provided. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effect of Grape Seed Proanthocyanidin (GSP) on fat metabolism and adipocytokines in obese rats. Fifty 5-week-old rats were randomly assigned to five groups (n = 10 per group) and given either a basal diet, a high-fat diet, or a high-fat diet supplemented with GSP (25, 50, and 100 mg/d) per group. The experiment lasted for five weeks, including a one-week adaptation period and a four-week treatment period. At the end of the experimental period, serum and adipose tissue samples were collected and analyzed. Additionally, we co-cultured 3T3-L1 preadipocytes with varying concentrations of GSP to explore its effect on adipocyte metabolism. The results demonstrated that GSP supplementation reduced weight, daily gain, and abdominal fat weight coefficient (p < 0.05). It also decreased levels of glucose, cholesterol (TC) (p < 0.05), triglycerides (TG) (p < 0.05), low-density lipoprotein (LDL), cyclooxygenase-2 (COX-2), and interleukin-6 (IL-6) in adipose tissue. Furthermore, GSP addition caused adipocyte crumpling in vitro and reduced the mRNA expression of COX-2, LEP, and TNF-α in adipocytes in vitro. These findings provide compelling evidence for exploring the role of GSP in the prevention and treatment of obesity and related diseases.

Abnormal adipocyte growth, distinguished by an increase in cell numbers and cellular differentiation, is regarded as a major pathological characteristic of obesity. Thus, inhibition of adipogenic differentiation in adipocytes could prevent obesity. Recently, certain probiotic stains have been reported to regulate lipid metabolism in vitro and/or in vivo. In this backdrop, this study aimed to investigate basic probiotic properties and potential antiobesity characteristics of mouse 3T3-L1 preadipocytes. Six lactic acid bacteria (LAB) strains were prescreened for their cholesterol-lowering activity, antioxidant activity, and survival at low pH and in a solution containing bile salts. These six strains were investigated for antiadipogenic activity by employing 3T3-L1 mouse preadipocytes. 3T3-L1 cells were treated with selected strains during the differentiation process. Lactobacillus johnsonii 3121 and Lactobacillus rhamnosus 86 were found to be more capable of reducing triglyceride and lipid accumulation, as compared to control group, which are fully differentiated 3T3-L1 adipocytes. These strains also inhibited adipocyte differentiation by downregulating the adipogenic transcription factor in 3T3-L1 adipocytes. Taken together, these results indicate that L. johnsonni 3121 and L. rhamnosus 86 could potentially act as probiotic bacteria and prevent fat accumulation by regulating adipogenesis-related markers. [ABSTRACT FROM AUTHOR]

E Se tea, processed by the tender leaves of Malus toringoides (Rehd.) Hughes, has been traditionally used as healthy tea to prevent hyperglycemic, hyperlipidemia and hypertensive disease. This paper firstly performed to explore its bioactive compounds and the possible protective mechanism on lipid accumulation. As a result, 13 flavonoids were isolated and identified, and phlorizin (2) was the most abundant compound in E Se tea. All the flavonoids could reduce TG level in 3T3-L1 preadipocytes. Oil red O staining results also verified that these flavonoids could inhibit the production of lipid droplet. Among them, compounds 2 and 6 have the strongest inhibitory effects on lipid accumulation. In addition, the synergistic effects between compound 2 and other compounds were further evaluated. Compared with individual flavonoid, most of combinations significantly reduced lipid accumulation, especially compounds 2 and 6. Western blotting analysis suggested compounds 2 and 6 could promote the phosphorylation of AMPK and ACC proteins. Furthermore, molecular docking results suggested that compounds 2 and 6 tightly bonded to AMPK by hydrogen bonding and electrostatic interaction. All the results suggested there was a significant synergistic effect between compounds 2 and 6 for inhibiting the lipid accumulation in 3T3-L1 preadipocytes. Therefore, E Se tea could be served as healthy tea to regulate lipid metabolism. [Display omitted] • The 13 Flavonoids were isolated and identified from the extract of E Se tea. • The inhibitory effects of 13 flavonoids on lipid accumulation were evaluated in adipocytes. • Synergistic inhibitory effects of flavonoid combinations were evaluated on lipid accumulation. • Molecular docking technology and western blotting were determined the mechanism. [ABSTRACT FROM AUTHOR]

Tetrabromobisphenol A (TBBPA), a widely-used brominated flame retardant, has been revealed to exert endocrine disrupting effects and induce adipogenesis. Given the high structural similarities of TBBPA analogues and their increasing exposure risks, their effects on lipid metabolism are necessary to be explored. Herein, 9 representative TBBPA analogues were screened for their interference on 3T3-L1 preadipocyte adipogenesis, differentiation of C3H10T1/2 mesenchymal stem cells (MSCs) to brown adipocytes, and lipid accumulation of HepG2 cells. TBBPA bis(2-hydroxyethyl ether) (TBBPA-BHEE), TBBPA mono(2-hydroxyethyl ether) (TBBPA-MHEE), TBBPA bis(glycidyl ether) (TBBPA-BGE), and TBBPA mono(glycidyl ether) (TBBPA-MGE) were found to induce adipogenesis in 3T3-L1 preadipocytes to different extends, as evidenced by the upregulated intracellular lipid generation and expressions of adipogenesis-related biomarkers. TBBPA-BHEE exhibited a stronger obesogenic effect than did TBBPA. In contrast, the test chemicals had a weak impact on the differentiation process of C3H10T1/2 MSCs to brown adipocytes. As for hepatic lipid formation test, only TBBPA mono(allyl ether) (TBBPA-MAE) was found to significantly promote triglyceride (TG) accumulation in HepG2 cells, and the effective exposure concentration of the chemical under oleic acid (OA) co-exposure was lower than that without OA co-exposure. Collectively, TBBPA analogues may perturb lipid metabolism in multiple tissues, which varies with the test tissues. The findings highlight the potential health risks of this kind of emerging chemicals in inducing obesity, non-alcoholic fatty liver disease (NAFLD) and other lipid metabolism disorders, especially under the conditions in conjunction with high-fat diets. [Display omitted] • TBBPA-BHEE, TBBPA-MHEE, TBBPA-BGE, and TBBPA-MGE could promote 3T3-L1 adipogenesis. • TBBPA analogs had a weak impact on the differentiation of C3H10T1/2 MSCs to brown adipocytes. • TBBPA-MAE promoted TG accumulation in HepG2 cells in the presence or absence of OA. • TBBPA-BHEE and TBBPA-MAE showed stronger obesogenic effects than TBBPA. • TBBPA analogs may induce tissue-specific perturbation in lipid metabolism. [ABSTRACT FROM AUTHOR]

Abstract Aims/Introduction The putative tumor suppressor gene, KBTBD11, might play a role in tumorigenesis, and is associated with cellular apoptosis and proliferation in colorectal cancer cells. However, the function of Kbtbd11 during adipogenesis is unknown. The aim of the present study was to investigate the role of Kbtbd11 in the differentiation of 3T3‐L1 preadipocytes. Materials and Methods For the fasting–refeeding protocol, mice were subjected to fasting for 24 h, followed by a chow diet for 12 h. Adenovirus infection methods were used to examine the effect of Kbtbd11, and 3T3‐L1 cells were analyzed with Oil Red O staining and real‐time polymerase chain reaction. Results The white adipose tissue expression of Kbtbd11 messenger ribonucleic acid (mRNA) was significantly higher in the re‐fed state than in the fasted state. Kbtbd11 mRNA levels were markedly increased in epididymal white adipose tissue of diet‐induced obesity mice compared with those in the mice fed a chow diet. In addition, Kbtbd11 mRNA expression was increased in a differentiation‐dependent manner in 3T3‐L1 cells. Knockdown of Kbtbd11 mRNA through the infection with adenoviral vectors remarkably inhibited triglyceride accumulation and adipocyte differentiation in 3T3‐L1 cells. In contrast, the overexpression of Kbtbd11 promoted the differentiation of 3T3‐L1 adipocytes. Conclusions The present findings show that Kbtbd11 expression might be involved in nutritional regulation and is increased in obese adipose tissue. In addition, Kbtbd11 appears to be required for the differentiation of adipocytes in 3T3‐L1 cells. Collectively, these results show a novel link between the expression of Kbtbd11 and fat accumulation, and suggest that Kbtbd11 is a new therapeutic target for obesity.

Rooibos (Aspalathus linearis) is a South African fynbos plant, well-known for its strong anti-oxidant capacity and use in cosmetic products. The anti-ageing properties of fermented and green (unfermented) rooibos were investigated using a cell culture model designed to evaluate the involvement of mitochondrial dysfunction in the age-related decline in preadipocyte function. Mitochondrial dysfunction was induced through long-term exposure of 3T3-L1 preadipocytes to ethidium bromide (EtBr). Mitochondrial DNA (mtDNA) depleted (ρ0) 3T3-L1 preadipocytes showed a significantly reduced rate of proliferation, delayed cell cycle progression (G0/G1 phase arrest), decreased mitochondrial membrane potential (ΔΨm), and increased glucose utilization and lactate production. Treatment with rooibos stimulated cell growth, attenuated G0/G1 phase arrest, improved ΔΨm, and increased glucose utilization and lactate production. Cellular ATP was significantly improved and AMPK activation was observed. The results obtained indicate that rooibos, particularly green rooibos, exhibit effects which preserve the functional capacity of preadipocytes exposed to ageing related insults.

Citrus maxima (Burm.) Merr (CMBM) is a traditional digestive tea in China, which is generally considered to have lipid-lowering effect. Here, a bioassay-guided identification of anti-obesity ingredients from CMBM was performed and four compounds were obtained, among which marmin exhibited excellent anti-proliferative activity on 3T3-L1 preadipocytes. Marmin could effectively promote the apoptosis of 3T3-L1 and inhibited its preadipocytes growth through induction of G2/M cell cycle arrest. The ROS levels and apoptotic bodies were increased after marmin treatment on 3T3-L1 preadipocytes, consistent with the results of flow cytometry. Moreover, The RT-PCR analysis revealed that marmin could significantly increase the mRNA expression levels of p53 and the ratio Bax/Bcl-2, but not the p21 mRNA expression. The Western Blotting results showed that marmin could suppress the phosphorylation of p65, extracellular signal- regulated protein kinase 1/2 (ERK1/2) and increase the phosphorylation of c-Jun N-terminal kinase (JNK), which indicated that marmin could induce 3T3-L1 preadipocytes apoptosis by inhibiting nuclear factor kappa-B (NF-κB) and altering mitogen-activated protein kinases (MAPKs) pathway.

Long non-coding RNAs (lncRNAs) have emerged as integral regulators of pathophysiological processes, but their specific roles and mechanisms in adipose tissue development remain largely unknown. Here, through microarray analysis, co-expression, and tissue specific analysis of adipocyte tissues after fasting for 72 h, we found that Lnc-FR332443 expression was dramatically decreased, as well as the expression of Runx1. The UCSC database and Ensembl database indicated that Lnc-FR332443 is the antisense lncRNA of Runx1. Lnc-FR332443 and Runx1 are highly enriched in adipose tissue and downregulated during adipogenic differentiation. Adipose tissue-specific knockdown of Lnc-FR332443 increased fat mass in vivo, and specific knockdown of Lnc-FR332443 in 3T3-L1 preadipocytes promoted adipogenic differentiation. In this process, Runx1 expression was decreased when Lnc-FR332443 was downregulated in adipocytes or 3T3-L1 preadipocytes, and vice versa, when Lnc-FR332443 was upregulated, the expression of Runx1 was increased. However, overexpression of Runx1 decreased the expression of the adipocyte cell marker genes PPARγ, C/EBPα and FABP4 significantly, while not affected the expression of Lnc-FR332443. Mechanistically, Lnc-FR332443 positively regulates Runx1 expression in mouse adipocytes and suppresses adipocyte differentiation by attenuating the phosphorylation of MAPK-p38 and MAPK-ERK1/2 expression. Thus, this study indicated that Lnc-FR332443 inhibits adipogenesis and which might be a drug target for the prevention and treatment of obesity.

Purposes: Nuciferine, a main aporphine alkaloid component found in lotus leaf (Nelumbo nucifera), has been demonstrated to possess the property of reducing fat mass and alleviating dyslipidemia in vivo. The purpose of this study is to explore the effects of nuciferine on the proliferation and differentiation of 3T3-L1 cells and further investigate the possible underlying molecular mechanisms.Methods: 3T3-L1 preadipocytes were treated with 0∼20 μM nuciferine for 24∼120 h, the cell viability was assessed using CCK8. 3T3-L1 preadipocytes and human primary preadipocytes were then induced differentiation and the effects of nuciferine on the lipid metabolism in differentiating and fully differentiated adipocytes were observed by the methods of intracellular triglyceride (TG) assay, Oil Red O staining, RT-qPCR and western blot. Transient transfection and dual luciferase reporter gene methods were used to assess the effects of nuciferine on FAS promoter activities.Results: Nuciferine inhibited the proliferation of 3T3-L1 preadipocytes in a dose- and time-dependent manner. 20 μM nuciferine significantly attenuated lipid accumulation and reduced intracellular TG contents by 47.2, 59.9 and 55.4% on the third, sixth and ninth day of preadipocytes differentiation, respectively (all p < 0.05). Moreover, the mRNA levels of PPARγ, C/EBPα, C/EBPβ, FAS, ACC, HSL and ATGL were notably decreased by 39.2∼92.5% in differentiating preadipocytes when treated with 5∼20 μM nuciferine (all p < 0.05). In fully differentiated adipocytes treated with 20 μM nuciferine for 48 h, the mRNA levels of FAS, ACC and SREBP1 were remarkably downregulated by 22.6∼45.2% compared with the controls (0 μM) (all p < 0.05), whereas the expression of adipokines FGF21 and ZAG were notably promoted by nuciferine. Similarly, in fully differentiated human primary adipocytes, the mRNA levels of FAS, ACC, SREBP1 were decreased and the expression of FGF21 and ZAG were elevated after treated with nuciferine (all p < 0.05). Further mechanism studies showed that 2.5∼20 μM nuciferine significantly decreased FAS promoter activities in 3T3-L1 preadipocytes.Conclusion: Nuciferine inhibited the proliferation and differentiation of 3T3-L1 preadipocytes. The inhibitory effects of nuciferine on adipogenesis might be due to the downregulation of PPARγ, C/EBPα and C/EBPβ, which led to the reduction of intracellular lipid accumulation in 3T3-L1 cells and by downregulating the expression of critical lipogenic enzymes, especially of FAS, which was achieved by inhibiting the FAS promoter activities. Besides, nuciferine promoted the expression of adipokines in fully differentiated adipocytes.

Maternal body mass index is associated with breast milk (BM) fatty acid composition. This study investigated the effects of BM omega (n)-6:n-3 polyunsaturated fatty acids (PUFAs) from non-obese women and women with obesity on the process of adipogenesis in 3T3-L1 preadipocytes. BM samples were collected from non-obese women (BMNO) and women with obesity (BMO) at one month postpartum. The fatty acid composition was measured, and BMNO and BMO groups with the lowest (Q1) and highest (Q4) quartiles of n-6:n-3 PUFA ratios were identified. 3T3-L1 preadipocytes were differentiated in the presence or absence of BM. Lipid accumulation and the expression of genes involved in lipogenesis and lipolysis were measured. Treatment with BMNO containing high (vs. low) n-6:n-3 PUFA ratios significantly increased the mRNA expression of lipogenic genes (acetyl-CoA carboxylase, fatty acid synthase, and stearoyl-CoA desaturase); however, there was no effect when cells were treated with BMO (with either low or high n-6:n-3 PUFA ratios). Treatment with BMO (high n-6:n-3 PUFA ratio) caused larger lipid droplets. Our findings demonstrated that BMNO with a high n-6:n-3 PUFA ratio was associated with a higher expression of lipogenic genes, while BMO with a high n-6:n-3 PUFA ratio showed larger lipid droplets, suggesting adipocyte dysfunction. These findings may have implications in the BM-mediated programming of childhood obesity.

Stevioside, the primary sweetener in stevia, is a glycoside with numerous beneficial biological activities. However, its anti-adipogenic effects on tissue differentiation and adipose tissues remain to be thoroughly investigated. In this study, the anti-adipogenic effects of stevioside during the differentiation of 3T3-L1 cells and epididymal adipose tissues of db/db mice were investigated by measuring the lipid droplets stained with Oil Red O and an immunoblot assay. Immunoblot analysis revealed that stevioside downregulated the expression of peroxisome proliferator-activated receptor-gamma (PPARγ), sterol regulatory element-binding protein-1c (SREBP-1c), CCAAT/enhancer-binding protein alpha (C/EBPα), and fatty acid synthase (FAS). Additionally, the protein expression of carnitine palmitoyltransferase 1 (CPT1), silent mating type information regulation 2 homolog 1 (SIRT1), and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) increased following treatment with stevioside. Furthermore, stevioside increased the phosphorylation of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), both in vitro and in vivo. The activity of AMPK in stevioside-treated 3T3-L1 cells was further confirmed using agonists and antagonists of AMPK signaling. Our data indicate that stevioside ameliorates anti-adipogenic effects and promotes β-oxidation in adipocytes by activating AMPK-mediated signaling. The results of this study clearly demonstrated the inhibitory effect of stevioside on the differentiation of adipocytes and the reduction of lipid accumulation in the epididymal adipose tissues of db/db mice.

The present study aimed to investigate the function and mechanisms of PAR2 in preadipocyte differentiation. This study found that the expression level of PAR2 was increased during 3T3-L1 mouse preadipocyte differentiation towards adipocytes. In addition, PAR2 overexpression significantly stimulated the expression of adipogenic proteins including ACC1, PPARγ, and SREBF1. Moreover, PAR2 overexpression increased the content of triglyceride (TG) in 3T3-L1 preadipocytes. Knockdown of PAR2 suppressed 3T3-L1 preadipocyte differentiation and adipogenesis. Mechanistically, PAR2 promoted 3T3-L1 preadipocyte differentiation and TG production through activation of the PI3K/AKT signalling pathway and MAT2A gene expression. The research sheds light on the adipogenic effects of PAR2 and its underlying mechanisms. Thus, PAR2 may have therapeutic significance for obesity. [ABSTRACT FROM AUTHOR]

Purpose: Inhibition of Notch signaling has been recently demonstrated to promote beige adipocyte biogenesis. However, most γ-secretase inhibitors (GSIs) used to achieve pharmacological inhibition of Notch signaling are at the basic research or preclinical stage, limiting the translation of fundamental findings into clinical practice. This present study aimed to evaluate the potential of several clinical candidates of GSIs as browning agents for the treatment of obesity. Methods: Seven GSIs that are clinical candidates for the treatment of Alzheimer's disease or cancer were selected and their impacts on Notch inhibition as well as promoting beige biogenesis were compared using in vitro culture of 3T3-L1 preadipocytes. Results: Four compounds (i.e.RO4929097, PF-03084014, LY3039478, and BMS-906024) that efficiently inhibited the expression of Notch target genes in 3T3-L1 preadipocytes were identified. Moreover, these compounds were optimized for dose-dependent effects at three gradient concentrations (0.5, 1, and 10 μM) to promote beige adipogenesis and mitochondrial biogenesis in 3T3-L1 preadipocytes without causing severe cytotoxicity. Conclusions: Our findings not only highlight the potential of cross-therapeutic application of these GSIs for obesity treatment via inhibition of γ-secretase-mediated processing of Notch signaling, but also provide important experimental evidence to support further design and development of clinically translatable Notch-inhibiting drug delivery systems. [ABSTRACT FROM AUTHOR]

Background: The incidence of obesity has soared over the last several decades. There is mounting evidence suggesting that the increased presence of environmental endocrine disruptors (EEDs), including nonylphenol (NP), plays an important role in the incidence of lipid metabolism disorders. The aim of this work was to determine whether chronical exposure to NP could induce obesity and lipid metabolism disorders, both in vivo in Sprague–Dawley rats, and in vitro in 3T3-L1 preadipocytes. Forty rats (n = 10 per group) were gavaged with NP in corn oil at dose levels of 0.02 μg/kg/day (low dose, L), 0.2 μg/kg/day (middle dose, M), and 2.00 μg/kg/day (high dose, H) or corn oil alone (vehicle control, C) for 180 days. In vitro study, 3T3-L1 preadipocytes were exposed to NP at concentrations of 0, 40 pM, 40 nM, or 40 μM for 12 days. Results: In vivo, the fat weight (F = 103.605, P < 0.001) and fat coefficient (F = 169.807, P < 0.001) of NP-exposed rats were higher than those of control group rats. The serum levels of TC (F = 3.798, P < 0.05), LDL-C (F= 4.946,P < 0.05), and TG (F = 14.117,P < 0.05) in the H group were higher than those in the control group. Protein concentrations of CEBPα (F = 189.104, P < 0.001), FAS (F = 51.011, P < 0.001), PPARγ (F = 114.306,P < 0.001), and SREBP1 (F = 30.432,P < 0.001) in serum in the NP group were higher than those in the control group. The concentration of NP in adipose tissues of rats increased with an increase in NP exposure dose in a dose–response manner (F = 561.353,P < 0.001). The numbers of adipocytes in the M and H groups decreased, and the volume of a single cell increased with cells' membranes ruptured. With the increase in NP exposure dose, the number of adipocytes per microscope decreased gradually (F = 85.873, P < 0.001). The expression levels of PPARγ (F = 169.936, P < 0.001) and FAS (F = 295.249, P < 0.001) proteins in the H group were higher than those in the control group. CEBPα (F = 101.086, P < 0.001) mRNA expression was up-regulated in the M and H groups; and FAS (F = 439.600, P < 0.001), PPARγ (F = 10.540, P < 0.001), and SREBP1 (F = 123.499, P < 0.001) mRNA expression in NP-exposed groups were significantly higher than those in the control group. In vitro, compared with the control group, the Oil Red Staining of adipocytes in the NP groups was darker, the fat cells were more densely distributed, and some of them fused into large lipid droplets. Expressions of CEBPα (F = 539.103, P < 0.001), FAS (F = 715.740, P < 0.001), PPARγ(F = 114.783, P < 0.001), and SREBP1 (F = 139.600, P < 0.001) proteins in 3T3-L1 preadipocytes were higher in group exposed to 40 μM NP than those in the control group. Conclusions: The results of this in vivo and in vitro experiment were consistent, and both have demonstrated that NP exposure interfered with the expression of proteins and/or mRNAs of lipid metabolism-related regulators (CEBPα, FAS, SREBP1, PPARγ), promoted the proliferation and differentiation of adipocytes and intracellular accumulation of lipids, and eventually lead to blood lipid disorders and obesity in rats. [ABSTRACT FROM AUTHOR]

Withania somnifera (Solanaceae), well-known as ‘Indian ginseng’ or ‘Ashwagandha’, is a medicinal plant that is used in Ayurvedic practice to promote good health and longevity. As part of an ongoing investigation for bioactive natural products with novel structures, we performed a phytochemical examination of the roots of W. somnifera employed with liquid chromatography–mass spectrometry (LC/MS)-based analysis. The chemical analysis of the methanol extract of W. somnifera roots using repeated column chromatography and high-performance liquid chromatography under the guidance of an LC/MS-based analysis resulted in a new withanolide, withasomniferol D (1). The structure of the newly isolated compound was elucidated by spectroscopic methods, including one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) and high-resolution (HR) electrospray ionization (ESI) mass spectroscopy, and its absolute configuration was established by electronic circular dichroism (ECD) calculations. The anti-adipogenic activities of withasomniferol D (1) were evaluated using 3T3-L1 preadipocytes with Oil Red O staining and quantitative real-time polymerase chain reaction (qPCR). We found that withasomniferol D (1) inhibited adipogenesis and suppressed the enlargement of lipid droplets compared to the control. Additionally, the mRNA expression levels of adipocyte markers Fabp4 and Adipsin decreased noticeably following treatment with 25 μM of withasomniferol D (1). Taken together, these findings provide experimental evidence that withasomniferol D (1), isolated from W. somnifera, exhibits anti-adipogenic activity, supporting the potential application of this compound in the treatment of obesity and related metabolic diseases.

Background/Aims: The aim of this study was to determine the direct role of liraglutide (LG) in adipogenesis and lipid metabolism. Methods: Lipid accumulation was evaluated by oil red O staining, quantitative real-time PCR (qPCR) was performed to determine glucagon-like peptide 1 receptor (GLP-1R), fatty acid synthase (FASN) and adipose triglyceride lipase (ATGL) expression in 3T3-L1 preadipocytes, differentiated adipocytes and in adipose tissues from mice. The effects of LG on 3T3-L1 adipogenesis and lipid metabolism were analyzed with qPCR, Western Blotting, oil red O staining, immunohistochemistry (IHC) and immunofluorescence (IF). All measurements were performed at least three times. Results: LG increased the expression of differentiation marker genes and lipid accumulation during preadipocyte differentiation. In differentiated adipocytes, LG decreased FASN expression, and simultaneously led to CREB phosphorylation and ERK1/2 activation which were abolished by a GLP-1R antagonist, exendin (9-39). LG induced-FASN down-regulation was partially reversed by PKA and ERK1/2 inhibitors. Consistent with above in vitro findings, LG treatment significantly reduced FASN expression in visceral adipose tissues of ob/ob mice, and reduced body weight gain. Conclusion: LG promotes preadipocytes differentiation, and inhibits FASN expression in adipocytes. LG induced down-regulation of FASN is at least partially mediated by PKA and MAPK signaling pathways.

Phlorizin is a natural polyphenol with many bioactivities, however, its low water solubility limits its application. Phlorizin oxidation product 2 (POP2) has higher water solubility than phlorizin. However, reaction velocity and conversion rate of POP2 are low, and its bioactivity has not been elucidated. Thus, we have established a new method for preparation of POP2 with a purity of 96.8%, and the inhibitory activity of POP2 on 3T3-L1 preadipocytes was investigated. POP2 decreased adipocyte viability and inhibited the accumulation of intracellular triglycerides (TG). Furthermore, POP2 suppressed transcription of fatty acid synthase (FAS) and glucose transporter 4 (GLUT-4), led to reduce lipid accumulation. Also, POP2 down-regulated transcription and expression of PPAR γ, C/EBPα, and FAS, led to inhibition of adipocyte differentiation. An efficient method to produce POP2 was developed, and we demonstrated that POP2 reduced lipid accretion. These results are useful for potential applications of phlorizin in the food and pharmaceutical industries.

ObjectiveGhrelin is a 28-amino-acid peptide that regulates energy hemostasis, glucose and lipid metabolism. We aimed to explore the effects of ghrelin on the proliferation and differentiation of 3T3-L1 and human primary preadipocytes.Methods3-(4,5-Dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) spectrophotometry, Oil Red O staining, intracellular glycerol-3-phosphate dehydrogenase (G-3-PDH) assays and semiquantitative reverse transcription polymerase chain reaction were used to investigate the action of ghrelin.ResultsGhrelin (0.01–1000 ng/ml) significantly increased the numbers of 3T3-L1 cells, and the maximum stimulatory effect was observed with the 10 ng/ml ghrelin treatment for 24 h (p < 0.05). Ghrelin also promoted the proliferation of human primary preadipocytes from 24 h (p < 0.05) to 48 h (p < 0.05) at a concentration of 1000 ng/ml. Further investigation showed that IGF-1 levels were notably increased in ghrelin-treated 3T3-L1 and human preadipocytes, and IGF-1 antibody was capable to attenuate this stimulatory action of ghrelin (all p < 0.05). Additionally, ghrelin significantly suppressed the differentiation of 3T3-L1 and human primary preadipocytes; 10 ng/ml ghrelin notably downregulated G-3-PDH activities in 3T3-L1 cells on day 3 and in human cells from days 4 to 12 following differentiation (all p < 0.05), and the intracellular lipoprotein lipase mRNA levels were lower than that of the controls (p < 0.05). Further investigation showed that the mRNA levels of peroxisome proliferator-activated receptor γ2 (PPARγ2) and CCAAT/enhancer binding protein α (C/EBPs) were also suppressed in ghrelin-treated human differentiating adipocytes.ConclusionGhrelin promotes the proliferation of 3T3-L1 and human primary preadipocytes by increasing the expression of IGF-1. Ghrelin inhibits murine and human adipocyte differentiation by downregulating PPARγ2 and C/EBPα levels, consequently leading to decreased lipid accumulation and lipogenic enzymes expression.

Diabetes mellitus (DM) is a metabolic disorder with numerous symptoms categorized via serves hyperglycemia effect along with altered fat, protein and carbohydrate metabolism mainly resultant from defects in insulin action/secretion or both. The aim of the current experimental study was to comfort the neuroprotective effect of ganoderic acid against the streptozotocin (STZ)-induced type I diabetes mellitus in mice and explore the underlying mechanism. Differentiation of 3T3-L1 preadipocytes effect; hepatic and glucose consumption effect of ganoderic acid was estimated on HepG2 cell lines and peroxisome proliferator-activated receptor (PPAR). FFA content was estimated in adipose and hepatic tissues. Ganoderic acid induced the 3T3-L1 preadipocytes differentiation. The mRNA expression of PPAR was increased in the high glucose-treated group in HepG2 and ganoderic acid treatment down-regulated the mRNA expression of PPAR. Ganoderic acid exhibited the inhibitory effect of α-glucosidase and α-amylase. Ganoderic acid demonstrated the reduced blood glucose and increase insulin level and also reduced the free fatty in hepatic and adipose tissue. Histopathological study showed the enhancement of β-cells in ganoderic acid-treated mice. Finally, their prebiotic effects on gut microbiota were illustrated via enhancing the population of diabetes resistant bacteria and also reducing the quantity of diabetes sensitive bacteria. Ganoderic acid attenuated STZ induced T1DM in mice via inflammatory pathways. [ABSTRACT FROM AUTHOR]