Lipid Accumulation In 3T3-L1 Cells Research Articles

Wounding stress enhances the anti-obesogenic, anti-inflammatory, and antioxidant properties of carrots (Daucuscarota)

This study evaluated the health benefits of carrots (Daucus carota) subjected to post-harvest wounding stress, emphasizing their anti-obesogenic, anti-inflammatory, and antioxidant properties. Carrots were shredded and stored at 15 ± 2 °C for 48 h to induce wounding stress, leading to a significant increase in chlorogenic acid content, by over 112.5 % compared to control. The study utilized 3T3-L1 cell differentiation assays to evaluate the anti-obesogenic potential, monitoring changes in lipid accumulation and adipogenesis-related gene expression. The anti-inflammatory activity was determined in LPS-stimulated RAW 264.7 macrophages by measuring nitrite production, while the antioxidant capacity was assessed using the DCFH-DA assay in Caco-2 cells. Key findings include a notable reduction in lipid accumulation in 3T3-L1 cells with stressed carrot extracts, particularly at 166 μg/mL concentration, showing an 18 % decrease compared to the control. Furthermore, there was a significant alteration in the expression of adipogenesis-related genes, with a 73 % decrease in FAS expression and a 160 % increase in ATGL expression. The stressed carrot extracts at 125 μg/mL also exhibited enhanced anti-inflammatory activity, inhibiting nitrite production by 44.60 %, a 153.26 % increase over the lower concentration. Additionally, antioxidant activity in stressed carrot extracts increased by 77.15 % at 125 μg/mL. These results indicate that wounding stress significantly boosts the health-promoting properties of carrots, suggesting their potential as functional foods for combating obesity and metabolic syndrome-related diseases. The study highlights the value of post-harvest treatments in enhancing the nutraceutical quality of vegetables.

Ancistrocladus tectorius Extract Inhibits Obesity by Promoting Thermogenesis and Mitochondrial Dynamics in High-Fat Diet-Fed Mice.

Root extracts of Ancistrocladus tectorius (AT), a shrub native to China, have been shown to have antiviral and antitumor activities, but the anti-obesity effects of AT aerial parts, mainly the leaves and stems, have not been investigated. This study is the first to investigate the anti-obesity effects and molecular mechanism of AT 70% ethanol extract in 3T3-L1 adipocytes and high-fat diet (HFD)-fed C57BL/6J mice. Treatment with AT extract inhibited lipid accumulation in 3T3-L1 cells and decreased the expression of adipogenesis-related genes. AT extract also upregulated the mRNA expression of genes related to mitochondrial dynamics in 3T3-L1 adipocytes. AT administration for 12 weeks reduced body weight and organ weights, including liver, pancreas, and white and brown adipose tissue, and improved plasma profiles such as glucose, insulin, homeostasis model assessment of insulin resistance, triglyceride (TG), and total cholesterol in HFD-fed mice. AT extract reduced HFD-induced hepatic steatosis with levels of liver TG and lipogenesis-related genes. AT extract upregulated thermogenesis-related genes such as Cidea, Pgc1α, Ucp1, Prdm16, Adrb1, and Adrb3 and mitochondrial dynamics-related genes such as Mff, Opa1, and Mfn2 in brown adipose tissue (BAT). Therefore, AT extract effectively reduced obesity by promoting thermogenesis and the mitochondrial dynamics of BAT in HFD-fed mice.

Epigallocatechin gallate suppresses mitotic clonal expansion and adipogenic differentiation of preadipocytes through impeding JAK2/STAT3-mediated transcriptional cascades

BackgroundMitotic clonal expansion (MCE) is a prerequisite for preadipocyte differentiation and adipogenesis. Epigallocatechin gallate (EGCG) has been shown to inhibit preadipocyte differentiation. However, the exact molecular mechanisms are still elusive. PurposeThis study investigated whether EGCG could inhibit adipogenesis and lipid accumulation by regulating the cell cycle in the MCE phase of adipogenesis and its underlying molecular mechanisms. Method3T3-L1 preadipocytes were induced to differentiate by a differentiation cocktail (DMI) and were treated with EGCG (25–100 μM) for 9, 18, and 24 h to examine the effect on MCE, or eight days to examine the effect on terminal differentiation. C57BL/6 mice were fed a high-fat diet (HFD) for three months to induce obesity and were given EGCG (50 or 100 mg/kg) daily by gavage. ResultsWe showed that EGCG significantly inhibited terminal adipogenesis and lipid accumulation in 3T3-L1 cells and decreased expressions of PPARγ, C/EBPα, and FASN. Notably, at the MCE phase, EGCG regulated the cell cycle in sequential order, induced G0/G1 arrest at 18 h, and inhibited the G2/M phase at 24 h upon DMI treatment. Meanwhile, EGCG regulated the expressions of cell cycle regulators (cyclin D1, cyclin E1, CDK4, CDK6, cyclin B1, cyclin B2, p16, and p27), and decreased C/EBPβ, PPARγ, and C/EBPα expressions at MCE. Mechanistic studies using STAT3 agonist Colivelin and antagonist C188–9 revealed that EGCG-induced cell cycle arrest in the MCE phase and terminal adipocyte differentiation was mediated by the inhibition of JAK2/STAT3 signaling cascades and STAT3 (Tyr705) nuclear translocation. Furthermore, EGCG significantly protected mice from HFD-induced obesity, reduced body weight and lipid accumulations in adipose tissues, reduced hyperlipidemia and leptin levels, and improved glucose intolerance and insulin sensitivity. Moreover, RNA sequencing (RNA-seq) analysis showed that the cell cycle changes in epididymal white adipose tissue (eWAT) were significantly enriched upon EGCG treatment. We further verified that EGCG treatment significantly reduced expressions of adipogenic factors, cell cycle regulators, and p-STAT3 in eWAT. ConclusionEGCG inhibits MCE, resulting in the inhibition of early and terminal adipocyte differentiation and lipid accumulation, which were mediated by inhibiting p-STAT3 nucleus translocation and activation.

Structurally Diverse Bisbenzylisoquinoline Alkaloids with Antiadipogenic Activity through PPARγ Downregulation from the Embryo of Nelumbo nucifera Seeds.

Six undescribed and six known bisbenzylisoquinoline alkaloids were isolated from the embryo of Nelumbo nucifera seeds. Their structures were fully characterized by a combination of 1H, 13C NMR, 2D NMR, and HRESIMS analyses, as well as ECD computational calculations. The antiadipogenic activity of 11 alkaloids was observed in a dose-responsive manner, leading to the suppression of lipid accumulation in 3T3-L1 cells. Luciferase assay and Western blot analysis showed that the active alkaloids downregulated peroxisome proliferator-activated receptor gamma (PPARγ, a key antiadipogenic receptor) expression in 3T3-L1 cells. Analysis of the structure-activity relationship unveiled that a 1R,1'S configuration in bisbenzylisoquinoline alkaloids led to a notable enhancement in antiadipogenic activity. The resistance level against lipid accumulation highlighted a consistent pattern with the suppressive effect on the PPARγ expression. These activity results indicate that alkaloids from the embryo of N. nucifera seeds have a potential of antiobesity effects through PPARγ downregulation.

Optimization of cultivar, germination time and extraction for radish sprout extract with high sulforaphene content.

Cruciferous vegetable sprout has been highlighted as a promising functional material rich in bioactive compounds called isothiocyanates (ITCs) and it can be grown in very short periods in controlled indoor farms. However, because ITCs content depends on multiple factors such as cultivar, germination time and myrosinase activity, those variables need to be controlled during germination or extraction to produce functional materials enriched in ITCs. Sulforaphene (SFEN), an ITC found primarily in radishes (Raphanus sativus L.), exerts beneficial effects on obesity. However, the optimal germination and extraction conditions for radish sprout (RSP) to increase SFEN content remain unascertained, and the extract's anti-obesity effect has yet to be evaluated. The present study found that the SFEN content was highest in purple radish sprout (PRSP) among the six cultivars investigated. Optimal SFEN content occurred after 2 days of PRSP germination (2 days PRSP). To maximize the dry matter yield, total ITCs and SFEN contents in RSP extract, we found the optimal conditions for extracting PRSP [27.5 °C, 60 min, 1:75.52 solute/solvent (w/v), no ascorbic acid] using response surface methodology. Consistent with high SFEN content, 2 days PRSP extract significantly outperformed 3 days or 4 days PRSP extract in inhibiting lipid accumulation in 3T3-L1 cells. Moreover, 2 days PRSP extract suppressed adipogenesis and lipogenesis-related protein expression. Regarding the cultivar, germination time and extraction conditions, optimally produced PRSP extract contains high SFEN content and exerts anti-obesity effects. Thus, we suggest PRSP extract as a potent functional material for obesity prevention. © 2024 Society of Chemical Industry.

Deubiquitinase USP1 enhances CCAAT/enhancer-binding protein beta (C/EBPβ) stability and accelerates adipogenesis and lipid accumulation

Dysregulation of the ubiquitin-proteasome system has been implicated in the pathogenesis of several metabolic disorders, including obesity, diabetes, and non-alcoholic fatty liver disease; however, the mechanisms controlling pathogenic metabolic disorders remain unclear. Transcription factor CCAAT/enhancer binding protein beta (C/EBPβ) regulates adipogenic genes. The study showed that the expression level of C/EBPβ is post-translationally regulated by the deubiquitinase ubiquitin-specific protease 1 (USP1) and that USP1 expression is remarkably upregulated during adipocyte differentiation and in the adipose tissue of mice fed a high-fat diet (HFD). We found that USP1 directly interacts with C/EBPβ. Knock-down of USP1 decreased C/EBPβ protein stability and increased its ubiquitination. Overexpression of USP1 regulates its protein stability and ubiquitination, whereas catalytic mutant of USP1 had no effect on them. It suggests that USP1 directly deubiquitinases C/EBPβ and increases the protein expression, leading to adipogenesis and lipid accumulation. Notably, the USP1-specific inhibitor ML323—originally developed to sensitize cancer cells to DNA-damaging agents—decreased adipocyte differentiation and lipid accumulation in 3T3-L1 cells without cytotoxicity. Oral gavage of ML323 was administered to HFD-fed mice, which showed weight loss and improvement in insulin and glucose sensitivity. Both fat mass and adipocyte size in white adipose tissues were significantly reduced by ML323 treatment, which also reduced the expression of genes involved in adipogenesis and inflammatory responses. ML323 also reduced lipid accumulation, hepatic triglycerides, free fatty acids, and macrophage infiltration in the livers of HFD-fed mice. Taken together, we suggest that USP1 plays an important role in adipogenesis by regulating C/EBPβ ubiquitination, and USP1-specific inhibitor ML323 is a potential treatment option and further study by ML323 is needed for clinical application for metabolic disorders.

Biofortified Beverage with Chlorogenic Acid from Stressed Carrots: Anti-Obesogenic, Antioxidant, and Anti-Inflammatory Properties.

Using wounding stress to increase the bioactive phenolic content in fruits and vegetables offers a promising strategy to enhance their health benefits. When wounded, such phenolics accumulate in plants and can provide antioxidant, anti-inflammatory, and anti-obesogenic properties. This study investigates the potential of using wounding stress-treated carrots biofortified with phenolic compounds as a raw material to extract carrot juice with increased nutraceutical properties. Fresh carrots were subjected to wounding stress via slicing and then stored at 15 °C for 48 h to allow phenolic accumulation. These phenolic-enriched slices were blanched, juiced, and blended with orange juice (75:25 ratio) and 15% (w/v) broccoli sprouts before pasteurization. The pasteurized juice was characterized by its physicochemical attributes and bioactive compound content over 28 days of storage at 4 °C. Additionally, its antioxidant, anti-inflammatory, and anti-obesogenic potentials were assessed using in vitro assays, both pre- and post-storage. The results reveal that juice derived from stressed carrots (SJ) possessed 49%, 83%, and 168% elevated levels of total phenolics, chlorogenic acid, and glucosinolates, respectively, compared to the control juice (CJ) (p < 0.05). Both juices reduced lipid accumulation in 3T3-L1 cells and nitric oxide production in Raw 264.7 cells, without significant differences between them. SJ further displayed a 26.4% increase in cellular antioxidant activity. The juice's bioactive characteristics remained stable throughout storage time. In conclusion, the utilization of juice obtained from stressed carrots in a blend with orange juice and broccoli sprouts offers a promising method to produce a beverage enriched in bioactive compounds and antioxidant potential.

Lactiplantibacillus sp. D10-2: potential bacteria for eliminating bisphenol A and reducing BpA-induced lipid accumulation.

Bisphenol A (BpA) is an endocrine-disrupting substance commonly found in plastics and resins. It is reported that BpA exposure induces lipid accumulation in humans, similar to obesogenic compounds. The main objective of this study is to investigate the removal of BpA using Lactiplantibacillus sp. D10-2, and to examine its potential for reducing BpA-induced lipid accumulation in 3T3-L1 cell line model. The heat-dried cells of Lactiplantibacillus sp. D10-2 showed 69.7% removal efficiency for initial BpA concentration of 10μg/mL, which was 30.5% higher than the live cells. The absence of metabolites or intermediates in BpA removal studies indicates that the Lactiplantibacillus sp. D10-2 strain removed BpA by adsorption process. The hydrophobic interactions of heat-dried Lactiplantibacillus sp. D10-2 cells were observed to be higher with 33.7% compared to live cells (15.0%), suggesting a stronger ability to bind with BpA. Although the BpA binding onto Lactiplantibacillus sp. D10-2 was not affected by pH, it was confirmed that as the temperature increases, the binding ability got decreased due to mass transfer and diffusion of BpA molecules. Treatment with Lactiplantibacillus sp. D10-2 (0.1, 0.25, 0.5, 1%) reduced lipid accumulation by 61.7, 58.0, 52.7 and 60.4% in 3T3-L1 cells exposed with BpA. In addition, it was confirmed that Lactiplantibacillus sp. D10-2 treatment suppressed the protein expression levels of lipogenesis-related PPARγ and C/EBPα in 3T3-L1 cells. The results of the study suggest that the Lactiplantibacillus sp. D10-2 strain can remove BpA and reduce BpA-accelerated lipid accumulation in 3T3-L1 cells.

Peroxisome Proliferator Activated Receptor-γ Agonistic Compounds from the Jellyfish-Derived Fungus Cladosporium oxysporum.

In our search for peroxisome proliferator-activated receptor (PPAR) agonists, five undescribed compounds, namely two acyclic diterpenes (1 and 2; cladopsol A and cladopsol B), two sesquiterpenes (3 and 4; cladopsol C and cladopsol D), and one C21-ecdysteroid (5; cladopsol E), and 15 known compounds were isolated from the jellyfish-derived fungus - Cladosporium oxysporum. The structures of the undescribed compounds were defined using UV, NMR, HR-ESI-MS, and electronic circular dichroism (ECD) spectroscopy and a modified Mosher's method. Luciferase reporter assay and docking analysis suggested that cladopsol B may function as a PPAR-γ partial agonist with a potential antidiabetic lead which may evade the side effects of full agonists. Moreover, cladopsol B stimulated glucose uptake in HepG2 cells with an efficacy comparable to that of rosiglitazone, but with less side effect induced by lipid accumulation in 3T3-L1 cells. Therefore, cladopsol B could serve as a molecular skeleton in a study of advanced antidiabetic lead with less side effect.

Extract of Isatidis Radix Inhibits Lipid Accumulation in In Vitro and In Vivo by Regulating Oxidative Stress.

Isatidis Radix (IR), the root of Isatis tinctoria L. belonging to Brassicaceae, has been traditionally used as a fever reducer. Although some pharmacological effects, such as anti-diabetes, anti-virus, and anti-inflammatory, have been reported, there is no study on the anti-obesity effect of IR. This study used 3T3-L1 cells, human mesenchymal adipose stem cells (hAMSCs), and a high-fat diet (HFD)-induced obese mouse model to confirm the anti-adipogenic effect of IR. Intracellular lipid accumulation in 3T3-L1 cells and hAMSCs was decreased by IR treatment.IR extract especially suppressed reactive oxygen species (ROS) production through a cluster of differentiation 36 (CD36)-AMP-activated protein kinase (AMPK) pathway. Consequently, the expressions of peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT-enhancer-binding proteins alpha (C/EBPα), and fatty acid synthesis (FAS) were inhibited by IR extract. In addition, β-oxidation-related genes were also decreased by treatment of IR extract. IR inhibited weight gain through this cascade in the HFD-induced obese mouse model. IR significantly suppressed lipid accumulation in epididymal white adipose tissue (eWAT). Furthermore, the administration of IR extract decreased serum free fatty acid (FFA), total cholesterol (TC), and LDL cholesterol, suggesting that it could be a potential drug for obesity by inhibiting lipid accumulation.

Inhibition of lipid droplet accumulation by Solanumnigrum by suppressing adipogenesis and inducing lipolysis, thermogenesis and autophagy in 3T3‑L1 cells.

It has been reported that Solanum nigrum exhibits anti-obesity effects in animal models induced by a high-fat diet. However, research on how Solanum nigrum exerts its anti-obesity effects is currently limited. Thus, the present study focused on identifying the mechanism of action associated with the anti-obesity activity of Solanum nigrum aerial part (SNAP), which significantly inhibited the accumulation of lipid droplets in differentiating 3T3-L1 cells. Intracellular lipid accumulation in 3T3-L1 cells was analyzed by Oil-Red O staining and glycerol content was analyzed using an ELISA kit. In addition, changes in protein expression within 3T3-L1 cells were analyzed using western blot analysis. It decreased the expression level of adipogenic proteins such as CCAAT/enhancer-binding protein α, Peroxisome proliferator-activated receptor γ, fatty acid binding protein 4, and adiponectin. In addition, SNAP increased the expression levels of lipolytic proteins, such as adipose triglyceride lipase and hormone-sensitive lipase, while decreasing perilipin-1. The treatment of fully differentiated 3T3-L1 cells increased the free glycerol levels. SNAP treatment resulted in increased AMP-activated protein kinase phosphorylation and the expression levels of thermogenic proteins (peroxisome proliferator-activated receptor-γ coactivator 1-α, PR domain containing 16 and uncoupling protein 1) and an autophagic protein (LC3-II). Overall, these results suggested that SNAP inhibited lipid droplet accumulation by suppressing adipogenesis and promoting lipolysis, thermogenesis and autophagy.

Effect of chlorogenic acid on lipid metabolism in 3T3-L1 cells induced by oxidative stress

Chlorogenic acid (CGA), a phenolic acid compound found in a variety of natural plants, has strong antioxidant capacity. In this study, we aimed to investigate the effects of CGA on differentiation and lipid metabolism of 3T3-L1 cells, as well as its protective effect against cellular oxidative stress induced by H2O2. We found that CGA can reduce the accumulation of intracellular ROS and attenuate the production of inflammatory cytokines TNF-α under oxidative stress. CGA treatment significantly inhibited preadipocytes differentiation and lipid accumulation in 3T3-L1 cells. Our results suggest that CGA may regulate lipid metabolism by reducing the expression of differentiation-related genes and inhibiting triacylglyceride synthesis. The findings of this study support CGA as a potential therapeutic agent for obesity. The effects of CGA and its metabolites in vivo should be investigated to further support its application in functional foods.

In Vitro Evaluation of Antidiabetic Potential of Cleistocalyx nervosum var. paniala Fruit Extract.

Diabetes mellitus is a complex global public health condition. Medicinal plants are significant resources in the research of alternative new drug active compounds. Cleistocalyx nervosum var. paniala (C. nervosum) is an indigenous berry fruit widely grown in Southeast Asia. The fruit of C. nervosum exhibit various medicinal properties and health benefits. This study aimed to investigate antidiabetic properties of C. nervosum fruit extract by in vitro assays and in vitro models. C. nervosum fruit extracted using three different solvents (hexane, ethanol, and distilled water) were tested for α-amylase and α-glucosidase inhibitory activities, followed by glucose uptake in HepG2 and L6 myoblasts. Lipid accumulation in 3T3-L1 cells treated with C. nervosum fruit extracts was then examined. The results revealed that ethanolic extract of C. nervosum fruit showed better inhibition against α-amylase (IC50 of 0.42 μg/mL) and α-glucosidase (IC50 of 0.23 μg/mL) compared with other extracts. Furthermore, ethanolic extract showed higher glucose uptake potential than the standard antidiabetic drug, metformin, in HepG2 cells. The ethanolic extracts resulted in enhanced glucose utilization in L6 myoblasts compared to untreated control. All extractions showed no significantly increased lipid accumulation in 3T3-L1 cells compared to the untreated control cells. The investigation confirmed that the ethanolic extract exhibited the highest antidiabetic activity among all extracts. These results imply that C. nervosum fruit extract has antidiabetic properties and therefore they may be used as useful therapeutic agents for treating diabetes.

Gut bacteria-derived 3-phenylpropionylglycine mitigates adipocyte differentiation of 3T3-L1 cells by inhibiting adiponectin-PPAR pathway.

Gut microbiota provide numerous types of metabolites that humans cannot produce and have a huge influence on the host metabolism. Accordingly, gut bacteria-derived metabolites can be employed as a resource to develop anti-obesity and metabolism-modulating drugs. This study aimed to examine the anti-adipogenic effect of 3-phenylpropionylglycine (PPG), which is a glycine conjugate of bacteria-derived 3-phenylpropionic acid (PPA). The effect of PPG on preadipocyte-to-adipocyte differentiation was evaluated in 3T3-L1 differentiation models and the degree of the differentiation was estimated by Oil red O staining. The molecular mechanisms of the PPG effect were investigated with transcriptome analyses using RNA-sequencing and quantitative real-time PCR. PPG suppressed lipid droplet accumulation during the adipogenic differentiation of 3T3-L1 cells, which is attributed to down-regulation of lipogenic genes such as acetyl CoA carboxylase 1 (Acc1) and fatty acid synthase (Fasn). However, other chemicals with chemical structures similar to PPG, including cinnamoylglycine and hippuric acid, had little effect on the lipid accumulation of 3T3-L1 cells. In transcriptomic analysis, PPG suppressed the expression of adipogenesis and metabolism-related gene sets, which is highly associated with downregulation of the peroxisome proliferator-activated receptor (PPAR) signaling pathway. Protein-protein association network analysis suggested adiponectin as a hub gene in the network of genes that were differentially expressed genes in response to PPG treatment. PPG inhibits preadipocyte-to-adipocyte differentiation by suppressing the adiponectin-PPAR pathway. These data provide a potential candidate from bacteria-derived metabolites with anti-adipogenic effects.

Anti-Obesity Activity in 3T3-L1 Cells of Cornus officinalis Fruits Harvested at Different Times

Obesity, a metabolic disease linked to several other diseases, is emerging as a global problem. We determined the harvest time that maximized the anti-obesity effect by evaluating the inhibition of lipid accumulation in 3T3-L1 cells treated with fruits of Cornus officinalis. FS1 (pericarp + seeds, harvested 15 September) showed excellent anti-obesity activity (Oil Red O; 26.12 ± 1.37% vs. MDI) and inhibited the expression of PPAR-γ (0.05 ± 0.01 ford vs. MDI), C/EBPα (0.03 ± 0.02 ford vs. MDI), and C/EBPβ (0.33 ± 0.07 ford vs. MDI) at 200 μg/mL. The antioxidant activity of FS1 was also the most effective. In addition, FS1 contained higher levels of active ingredients than samples harvested in other periods. Especially, content of loganin, verbenalin, and sweroside was high. In terms of anti-obesity activity and functional components, immature COF (FS1) was the best, and these results indicate that it is necessary to adjust the harvest time, when used as an anti-obesity agent.

Pea albumin extracted from pea (Pisum sativum L.) seed protects mice from high fat diet-induced obesity by modulating lipid metabolism and gut microbiota

Plant protein has been reported to play a key role in the prevention of obesity and associated complications. It is unknown whether pea albumin may exert anit-obesity and obesity-associated metabolic disorders alleviation. Pea albumin was isolated from pea seed (Pisum sativum L.) and determined its functional role for anti-obesity and metabolic disorders alleviation in high fat diet (HFD)-fed mice. Pea albumin administration reduced body weight gain, improved glucose tolerance and insulin sensitivity, reduced inflammatory cytokines secretion, and alleviated hepatic steatosis in HFD-fed mice. Interestingly, pea albumin inhibited lipid accumulation in 3T3-L1 cells in vitro and modulated lipid metabolism by upregulating critical proteins implicated in lipolysis and fatty acid oxidation, and downregulating lipogenesis in vivo. Moreover, pea albumin restored gut microbial composition to normal fat diet condition and selectively promoted the growth of beneficial gut bacteria (Akkermansia, Parabacteroides etc.). Collectively, the data demonstrated that pea albumin protects mice from HFD-induced obesity and associated metabolic disorders.

Inhibitory Effects of Hydrolysable Tannins on Lipid Accumulation in 3T3-L1 Cells.

Obesity is currently the most common cause of metabolic diseases including type 2 diabetes and hyperlipidemia. Obesity results from excess lipid accumulation in adipose tissue. Several studies have investigated the inhibitory effects of natural plant-derived products on adipocyte differentiation and lipid accumulation. In this study, we examined the effect of hydrolysable tannins composed of gallic acid and glucose on adipocyte differentiation in 3T3-L1 cells. 1,2,3,4,6-Penta-O-galloyl-β-D-glucose (PGG) (1), a representative gallotannin, inhibited lipid accumulation in 3T3-L1 cells, whereas ellagitannins (tellimagrandin I, eugeniin and casuarictin) did not. The expression of adipocyte differentiation-related genes, including peroxisome proliferator activator γ2 (Pparγ2), CCAAT/enhancer binding protein α (C/EBPα) and adipocyte fatty acid binding protein (aP2), was significantly suppressed in PGG (1)-treated 3T3-L1 cells beginning at day 2 after induction of differentiation. While PGG (1) did not directly reduce Pparγ2 expression, it reduced the expression of its target genes in mature adipocytes. In addition, PGG (1) treatment inhibited mitotic clonal expansion, one of earliest events of adipocyte differentiation. These findings indicate that PGG (1) has an inhibitory effect on adipocyte differentiation through the suppression of mitotic clonal expansion.

DEPP Deficiency Contributes to Browning of White Adipose Tissue.

Decidual protein induced by progesterone (DEPP) was originally identified as a modulator in the process of decidualization in the endometrium. Here, we define that DEPP is involved in adipose tissue thermogenesis, which contributes to metabolic regulation. Knockdown of DEPP suppressed adipocyte differentiation and lipid accumulation in 3T3-L1 cells, induced expression of brown adipose tissue (BAT) markers in primary brown adipocyte and induced mouse embryonic fibroblasts (MEFs) differentiation to brown adipocytes. Moreover, DEPP deficiency in mice induced white adipocyte browning and enhanced BAT activity. Cold exposure stimulated more browning of white adipose tissue (WAT) and maintained higher body temperature in DEPP knockout mice compared to that in wild-type control mice. DEPP deficiency also protected mice against high-fat-diet-induced insulin resistance. Mechanistic studies demonstrated that DEPP competitively binds SIRT1, inhibiting the interaction between peroxisome proliferator-activated receptor gamma (PPARγ) and Sirtuin 1 (SIRT1). Collectively, these findings suggest that DEPP plays a crucial role in orchestrating thermogenesis through regulating adipocyte programs and thus might be a potential target for the treatment of metabolic disorders.

Plant miR167e-5p promotes 3T3-L1 adipocyte adipogenesis by targeting β-catenin.

Adipogenesis is important in the development of fat deposition. Evidence showed that plant microRNAs (miRNAs) could be absorbed by the digestive tract and exert regulatory effects on animals’ physiological processes. However, the regulation of plant miRNA on host lipogenesis remains unknown. This study explored the potential function of plant miRNA, miR167e-5p, in adipogenesis in vitro. The presentation of plant miR167e-5p improved lipid accumulation in 3T3-L1 cells. Bioinformatics prediction and luciferase reporter assay indicated that miR167e-5p targeted β-catenin. MiR167e-5p could not only negatively affect the expression of β-catenin but also showed a positive effect on several fat synthesis–related genes, peroxisome proliferator–activated receptor gamma (Pparγ), CCAAT/enhancer-binding protein α (Cebpα), fatty acid-binding protein 4 (Ap2), lipolysis genes, adipose triglyceride lipase (Atgl), and hormone-sensitive lipase (Hsl) messenger RNA levels. Meanwhile, lipid accumulation and the expression of the β-catenin and other five fat synthesis–related genes were recovered to their original pattern by adding the miR167e-5p inhibitor in 3T3-L1 cells. The immunoblot confirmed the same expression pattern in protein levels in β-catenin, PPAR-γ, FAS, and HSL. This research demonstrates that plant miR167e-5p can potentially affect adipogenesis through the regulation of β-catenin, suggesting that plant miRNAs could be a new class of bioactive ingredients in adipogenesis.

Isolation of phenolic acids and tannin acids from Mangifera indica L. kernels as inhibitors of lipid accumulation in 3T3-L1 cells.

Mango (Mangifera indica L.) kernels are usually discarded as waste, but they contain many pharmacological properties and bioactivities. In this study, we isolated antiobesity agents from mango kernels that inhibit intracellular lipid formation in 3T3-L1 adipocytes. Two phenolic acids, ethyl gallate and ethyl digallate, and 2 tannin acids, 1,2,3,4,6-penta-O-galloyl-β-d-glucose (PGG) and 3-O-digalloyl-1,2,4,6-tetra-O-β-d-glucose (HGG), were identified from mango kernels and were found to be suppressed lipid accumulation as evidenced by Oil Red O staining. Furthermore, ethyl digallate, PGG, and HGG significantly downregulated the mRNA expression of adipogenic transcription factors such as C/EBPα and PPARγ. However, ethyl gallate did not affect the expression of these transcription factors. Our findings reveal the presence of antiobesity compounds in mango kernels, implying its therapeutic role against obesity.