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Plant seed oils are rich in unsaturated fatty acids and lipid-soluble active ingredients, which have become a hot spot of research in recent years. In this study, a novel low-temperature continuous phase transition extraction device was utilized to extract passion fruit seed oil and its lipid composition, quality properties and hypolipidemic activity were investigated. The extraction rate of passion fruit seed oil by low-temperature continuous phase transition extraction was 20.37 %, which was about twice as much as that of solvent extraction (10.80 %). The contents of unsaturated fatty acids, vitamin E and β-carotene in passion fruit seed oil extracted by low-temperature continuous phase transition were 84.00 %, 21.03 μg/g and 6.28 μg/g, respectively, which were significantly better than those of solvent extraction. The acid value, peroxide value and saponification value of the resulting passion fruit seed oil were 0.275 mg/g, 0.034 g/100g and 185.175 mg/g, respectively. Passion fruit seed oil exhibited good hypolipidemic activity, and the IC50 of its inhibitory effect on pancreatic lipase and cholesterol esterase were 97.19 μg/mL and 48.08 μg/mL, respectively. Further 3T3-L1 cell experiments concluded that passion fruit seed oil not only reduces lipid production in cells, but also reduces triglyceride, total cholesterol, and low-density lipoprotein cholesterol content in cells, while elevating high-density lipoprotein cholesterol content. This study provides a theoretical basis for the development and utilization of passion fruit seed oil as a functional lipid.

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.

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.

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.

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.

A comparative transcriptomic analysis in adipose stem and progenitor cells (ASPCs) between obese and lean mice might facilitate the identification of novel adipogenic genes. Here, we compare transcriptomic differences in the ASPCs of subcutaneous adipose tissue (SAT) between the mice fed on a high-fat-diet (HFD) and the chow diet (CD)-fed mice by analyzing three independent single-cell RNA sequencing datasets. Six differential genes, including three up-regulated genes Aspn, Rrbp1, Fbln2 and three down-regulated genes Htra1, Plpp3, Efemp1, are identified and confirmed in HFD-fed mice. Further, the expression of these genes is found to be significantly diminished in the differentiated 3T3-L1 cells. Treatment with luteolin, a dietary flavonoid known to inhibit 3T3-L1 adipogenesis, reverses the decreased expression of Aspn, Htra1 and Efemp1. Furthermore, knockdown of Aspn, Htra1 and Efemp1 significantly facilitates 3T3-L1 adipogenesis. Together, these genes not only are differential in ASPCs between obese and lean mice, but also are the adipogenic inhibitory genes that can be up-regulated by luteolin treatment., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024. Published by Elsevier B.V.)

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.

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.

Tetrabromobisphenol A (TBBPA) and tetrabromobisphenol S (TBBPS) are widely distributed brominated flame retardants. While TBBPA has been demonstrated to stimulate adipogenesis, TBBPS is also under suspicion for potentially inducing comparable effects. In this study, we conducted a non-targeted metabolomics to examine the metabolic changes in 3T3-L1 cells exposed to an environmentally relevant dose of TBBPA or TBBPS. Our findings revealed that 0.1 µM of both TBBPA and TBBPS promoted the adipogenesis of 3T3-L1 preadipocytes. Multivariate analysis showed significant increases in glycerophospholipids, sphingolipids, and steroids relative levels in 3T3-L1 cells exposed to TBBPA or TBBPS at the final stage of preadipocyte differentiation. Metabolites set composed of glycerophospholipids was found to be highly effective predictors of adipogenesis in 3T3-L1 cells exposed to TBBPA or TBBPS (revealed from the receiver operating characteristic curve with an area under curve > 0.90). The results from metabolite set enrichment analysis suggested both TBBPA and TBBPS exposures significantly perturbed steroid biosynthesis in adipocytes. Moreover, TBBPS additionally disrupted the sphingolipid metabolism in the adipocytes. Our study presents new insights into the obesogenic effects of TBBPS and provides valuable information about the metabolites associated with adipogenesis induced by TBBPA or TBBPS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Daidzein is one of the main bioactive substances of soybean isoflavones that has a wide range of health benefits. This research explored the anti-obesity effect of daidzein and the mechanism of lipid metabolism for adipogenesis in 3T3-L1 adipocytes by experiment design of reducing matrix effects. The results indicated that high concentrations of daidzein significantly inhibited preadipocyte proliferation, fat deposition and the ratio of linolenic acid in 3T3-L1 adipocytes. Western blot and relative quantitative RT-PCR results indicated that daidzein significantly decreased protein and gene levels of the nuclear transcription factors C/EBPα and PPARγ. Meanwhile, daidzein also downregulated the expression of downstream genes (LPL, FAS, ACC, SCD1, FADS2); however, the relative expression levels were not dose-dependent for daidzein. Therefore, isolated from soybean, daidzein as a plant health product has a potential function for the prevention and therapy of obesity and related metabolic disorders.

Erucin is a natural isothiocyanate found in rocket salad and broccoli which are believed to have health benefits including weight control effect and used as a functional food globally. Though anti-cancer effect of erucin has been intensively studied, anti-obesity effect of erucin and its molecular mechanisms have not yet been reported. In this study, we investigated anti-adipogenic effect of erucin in 3T3-L1 preadipocytes and its underlying molecular mechanisms. Erucin inhibited adipogenesis of 3T3-L1 preadipocytes and reduced adipogenic gene expressions. This anti-adipogenic activity of erucin was most effective at early stage of adipogenesis including mitotic clonal expansion (MCE). Fluorescence-activated cell sorter (FACS) analysis and trypan blue assay results showed that erucin induced G1/S arrest and inhibited cell proliferation at MCE stage. Western blot analysis results showed that erucin inhibited extracellular signal-regulated kinase (ERK) signaling pathway by blocking Ras activity. Taken together, these results indicated that erucin inhibits adipogenesis of 3T3-L1 preadipocytes by blocking Ras activity at early stage of adipogenesis.

Coffee consumption has been shown to reduce the risk of developing type 2 diabetes mellitus (T2DM) in humans; however, the exact mechanism is not completely understood. Here, we demonstrate that N-caffeoyltryptophan (CTP), an ingredient of coffee, enhances adipogenic differentiation and promotes glucose uptake into adipocytes. CTP increased lipid accumulation and adipogenic markers (PPARγ, C/EBPα, and FABP4) expression in mouse 3T3-L1 preadipocyte cell lines and primary preadipocytes. In addition, CTP promoted glucose uptake in 3T3-L1 cells. In the oral glucose tolerance test, daily administration of CTP (30 mg/kg/day, i.p.) for a week reduced blood glucose levels in mice. In 3T3-L1 cells, adipogenic differentiation and increased adipogenic markers expression induced by CTP were inhibited by U0126, a selective MEK1/2 inhibitor. Furthermore, mRNA induction of Pparg by CTP was abrogated in SIRT1 siRNA-transfected 3T3-L1 cells. These results suggest the involvement of the MEK/ERK signaling and SIRT1 in the mechanism of adipogenic function of CTP. Taken together, CTP might contribute to the reduction in postprandial glycemia and a subsequent reduction in onset risk for T2DM., Competing Interests: Declaration of Competing Interest The authors have explicitly stated that there are no conflicts of interest in connection with this article., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Recent studies have shown that exposure to some organophosphates, such as triphenyl phosphate (TPHP) and diphenyl phosphate (DPHP), can affect adipogenesis in preadipocytes. 2-Ethylhexyl diphenyl phosphate (EHDPP), an organophosphate, is frequently detected in various environmental media. However, there is less information about the toxicity effects and the mechanism by which EHDPP affects preadipocytes. In the present study, we investigated whether EHDPP could induce differentiation in 3T3-L1 preadipocytes through the peroxisome proliferator-activated receptor γ (PPARγ) signaling pathway. The fluorescence competitive binding assay and the dual-luciferase reporter gene assay were used to assess the binding affinity and activation of PPARγ, and the results showed that EHDPP can bind to the ligand binding domain of PPARγ (PPARγ-LBD) and activate PPARγ in vitro. Exposure to EHDPP for 10 days extensively induced adipogenesis in 3T3-L1 preadipocytes as assessed by lipid accumulation and gene expression of adipogenic markers of fatty acid binding protein 4 (FABP4), lipoprotein lipase (Lpl), adiponectin (Adip), and fatty acid synthase (Fasn). Furthermore, the preadipocytes differentiation was blocked by the PPARγ-specific antagonist GW9662, indicating that the PPARγ signaling pathway plays an important part in 3T3-L1 cell differentiation induced by EHDPP. Taken together, EHDPP can bind to PPARγ-LBD, activate PPARγ receptor, and induce cell differentiation via the PPARγ signaling pathway in 3T3-L1 preadipocytes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The differentiation efficiency of 3T3-L1 preadipocytes is an essential factor affecting studies on cellular mechanisms associated with obesity, diabetes, and related disorders. Differentiation of 3T3-L1 cells is commonly induced by an adipogenic cocktail containing insulin, dexamethasone (DEX), and 3-isobutyl-1-methylxanthine (IBMX). However, 3T3-L1 cells after freezing and thawing for many times always have a low differentiation efficiency. To solve this problem, we compared the differentiation efficiency of six commonly used adipogenic cocktails and protocols published in 2017. On this basis, we further compared 18 adipogenic cocktails with 2 μM rosiglitazone added and/or with a prolonged treatment with IBMX. The results revealed that the adipogenic cocktail containing 0.5 mM IBMX, 1 μM DEX, and 10 μg/mL insulin was the most effective for the frozen-thawed 3T3-L1 cells differentiation. Rosiglitazone, and IBMX under a prolonged treatment, could improve the differentiation efficiency of the frozen-thawed 3T3-L1 cells. However, the effect was closely related to concentrations of agents in the adipogenic cocktails., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The leaves of Ilex Kudingcha, locally named "Kudingcha" in China, has been traditionally applied for treating obesity. Studies have demonstrated that the ethanol extract of Ilex kudingcha have anti-adipogenic effects. However, the constituent which was responsible for its anti-obesity and its underlying molecular mechanism has not yet been elucidated. This research explored the anti-obesity effect of kudinoside-D which was a main natural component of triterpenoid saponin from the ethanol extract of Ilex kudingcha, on lipid accumulation and the potential mechanism of action of adipogenesis in 3T3-L1 adipocytes. The adipocytes were treated with various concentrations of kudinoside D (0 to 40μM) during differentiation. The image-based Oil Red O staining analyses revealed that KD-D, dose dependently reduced cytoplasmic lipid droplet in 3T3-L1 adipocytes with the IC 50 is 59.49μM. Meanwhile, major adipogenic transcription factor peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα) and sterol regulatory element-binding protein 1c (SREBP-1c) were significantly repressed as well as their target genes. The phosphorylation of AMP-activated protein kinase (AMPK) and its downstream target phosphorylated-acetyl CoA carboxylase (ACC) expression were also increased. In addition, the inhibitory effects of KD-D on the expressions of PPARγ and C/EBPα were weakened when cells were cotreated with AMPK inhibitor Compound C. These results indicated KD-D exerts anti-adipogenic effects through modulation of adipogenic transcription factors via AMPK signaling pathway. And the current findings demonstrated that KD-D was a potential therapeutic candidate for alleviating obesity and hyperlipidemia., (Copyright © 2017. Published by Elsevier B.V.)

PU.1, an Ets family transcription factor, was previously demonstrated expressed in 3T3-L1 preadipocytes and had an negative effect on adiopogenesis. However, the underlying mechanism remains elusive. Here, miR-191 was identified as an inhibitor of adipocyte differentiation through targeting the 3' untranslated regions of C/EBPβ, the initial factor in the C/EBPα/β-PPARγ terminal pathway of adipogenic differentiation. MiR-191 suppressed the lipid accumulation by Oil Red O staining and downregulated the levels of adipogenic marker genes PPARγ (P<0.01), aP2 (P<0.01) and FAS (P<0.05). Then, we found that PU.1 overexpression resulted in upregulation of miR-191 and adipogenic inhibition. Likewise, PU.1 overexpression rescued the miR-191 decrease and resisted the adipogenic promotion caused by miR-191 oligonucleotide inhibitor. Collectively, these results revealed that PU.1 promoted miR-191 to suppress adipogenesis 3T3-L1 preadipocyte and indicated a new mechanism of PU.1 inhibiting adipogenesis., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Safflor yellow A (SY) has been demonstrated to be beneficial to cardiovascular system. Our previous study showed that hydroxysafflor yellow A (HSYA), a main component of SY, could increase peroxisome proliferator-activated receptor γ mRNA expression. In this study, we investigate the effect of HSYA on the proliferation and adipogenesis of mouse 3T3-L1 preadipocytes., Methods: The proliferation and adipogenesis of 3T3-L1 cells treated with HSYA was studied by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) spectrophotometry, Oil Red O staining and intracellular triglyceride assay methods. HSL mRNA expression and promoter activity were studied by real-time quantitative RT-PCR, transient transfection and dual luciferase reporter gene methods., Results: HSYA (0.1 mg/L) significantly inhibited the proliferation of 3T3-L1 cells when compared with control cells in 8 h. This effect was further enhanced with the extension time (24 to 96 h) and an increase of concentration of HSYA (1-10 mg/L). The maximal inhibitory action was observed at 0.1 mg/L HSYA in 72 h (86±11.8% vs. 100±4.1%, p<0.01). HSYA notably reduced the amount of intracellular lipid and triglyceride content in adipocytes to 85% (1 mg/L) and 75% (100 mg/L) on Day 4 following the differentiation, respectively, while increased HSL mRNA expression and promoter activities to 2.7 fold and 1.55 fold, respectively (p<0.01), in differentiated 3T3-L1 adipocytes., Conclusions: HSYA inhibits the proliferation and adipogenesis of 3T3-L1 preadipocytes. The inhibitory action of HYSA on adipogenesis may be due to the promotion of lipolytic-specific enzyme HSL expression by increasing HSL promoter activity., (Copyright © 2014 Elsevier B.V. All rights reserved.)