Vaccarin,a flavonoid glycoside isolated from Vaccaria segetalis,is non-toxic to 3T3-L1 cells up to concentrations of 200μM.Accordingly,we investigated the effects of this natural product on adipogenesis and lipolysis...Vaccarin,a flavonoid glycoside isolated from Vaccaria segetalis,is non-toxic to 3T3-L1 cells up to concentrations of 200μM.Accordingly,we investigated the effects of this natural product on adipogenesis and lipolysis in 3T3-L1 adipocytes.Our results revealed that vaccarin significantly inhibited lipid accumulation by suppressing the adipogenesis-related transcription factors peroxisome proliferator-activated receptorγ(PPARγ)and the CCAAT/enhancer-binding proteinα(C/EBPα).Specifically,lipid accumulation decreased by up to 27.7±2.7%when 3T3-L1 adipocytes were treated with a 10μM concentration of vaccarin.Mechanistic studies showed that the compound inhibited adipogenesis through activation of the Hedgehog(Hh)signaling pathway and so restoring Smo and Gli1 expression at an early stage of differentiation.In mature 3T3-L1 cells,vaccarin significantly increased the secretion of glycerol into the surrounding medium and thus indicating that it accelerated the degradation of triglycerides.In addition,vaccarin,was shown to enhance lipolysis through stimulation of the transcription levels of lipoprotein lipase,monoglycerides lipase,adipose triacylglyceride lipase,hormone-sensitive lipase and adipose differentiated-related protein.All told,vaccarin suppressed lipid accumulation and enhanced lipolysis during adipocyte differentiation by restoring Hh signaling.As such,it is a phytochemical capable of halting adipocyte hyperplasia and,thereby,ameliorating the effects of obesity.展开更多
Ligustri lucidi Fructus is a traditional Chinese medicine and possesses various bioactivities,including hypoglycemic effect.Ligustrum lucidum Ait flowers are poorly investigated.Thus,we hypothesized that L.lucidum flo...Ligustri lucidi Fructus is a traditional Chinese medicine and possesses various bioactivities,including hypoglycemic effect.Ligustrum lucidum Ait flowers are poorly investigated.Thus,we hypothesized that L.lucidum flowers also could have hypoglycemic effect.Chemical composition and glucose uptake effect of L.lucidum flowers on 3T3-L1 adipocytes were investigated.In this study,the components of L.lucidum flowers were investigated by various chromatographic and spectroscopic methods and the effects of L.lucidum flowers on the induction of glucose uptake were investigated by 3T3-L1 adipocytes.Seven compounds were isolated and identified from L.lucidum flowers,including ursolic acid(1),kaempferol-7-O-α-Lrhamnoside(2,KR),β-sitosterol(3),β-daucosterol(4),kaempferitrin(5,KF),10-hydroxy oleuropein(6,HO),and kaempferol-3-O-β-D-glucopyranoside-7-α-L-rhamnopyranoside(7,KGR).The results of glucose uptake showed that total extract(TE),KR,KF and HO from L.lucidum flowers significantly enhanced glucose uptake.This study demonstrated that compounds 2,5-7 were isolated for the first time from this plant and compounds 2 and 7 were isolated for the first time in genus Ligustrum,L.lucidum flowers,KR,KF and HO may possess potential hypoglycemic effect.展开更多
The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food...The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food to treat obesity and diabetes, The isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography. With berberine as a positive control, different concentrations ofsporamin （0.000, 0.125, 0.025, 0.250, 0.500, and 1.000 mg·mL^-1 were used to treat 3T3-L1 preadipocytes. Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry, Preadipocytes differentiation was measured by 3（4,5-dimethylthiazolyl-2-yl）-2,5-diphenyltetrazolium bromide （MTT） spectrophotometric assay. Two sporamin proteins, which were separated into sporamin A （31 kD） and sporamin B （22 kD）, could be purified by ion-exchange and gel filtration chromatography. After being treated by different concentrations of sporamin, the differentiation of 3T3-L1 preadipocytes was significantly inhibited, compared with the positive control. When the sporamin solution concentration was 0.500 mg mL-1, the accumulation of lipid droplets within the cells was significantly decreased and the optical density （OD） value of the solution from destained Oil Red O reached to 0.35, which was the lowest value （P〈 0.05）. The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations. In addition, the inhibitory effect was more obvious with the prolonged treatment time （P〈 0.05）. The differentiation and proliferation of 3T3-L1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin. It was, therefore, suggested that the sporamin was potentially effective for weight loss.展开更多
Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained wi...Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained with OilRed-O staining to confirm adipogenesis.The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay,intracellular reactive oxygen species generation through flow cytometry,and morphologically through confocal microscopy using propidium iodide,4,6-diamino-2-phenylindol dihydrochloride,and 2’,7’-dichlorofluorescein diacetate dyes.The uptake of fluorescent glucose analog,2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy.Results:MTT assay showed that the cell survival rate was(28.00±3.00)%,(92.33±2.60)%,and(71.34±2.10)%in the presence of 2 mM H2O2,AbPI alone,and AbPI and H2O2 both,respectively,in comparison to the control.Oil-Red-O staining indicated that Ab PI enhanced adipogenesis.AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone,and showed insulinsensitizing effect in the presence of insulin,but failed to stimulate the uptake in the absence of insulin.Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon Ab PI treatment.Confocal microscopy showed that the damaged cell population rose to 3.50%,117.84%,and 261.50%in the presence of Ab PI alone,AbPI with H2O2,and H2O2 alone,respectively.Conclusions:The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.展开更多
Objective:3,4-Oxo-isopropylidene-shikimic acid(ISA),a derivative of shikimic acid,has exhibited ameliorative effect on cognitive impairment in experimental animal models of dementia.This study investigated the effect ...Objective:3,4-Oxo-isopropylidene-shikimic acid(ISA),a derivative of shikimic acid,has exhibited ameliorative effect on cognitive impairment in experimental animal models of dementia.This study investigated the effect of ISA on lipid accumulation and adipokine secretion during differentiation of 3T3-L1 fibroblasts to adipocytes.Methods:3T3-L1 cells were cultured and treated with ISA(50e800 mM)from days 3e8.Lipid accumulation and triglyceride content were measured.Gene expression of adipokines(adiponectin,leptin,and resistin),CCAAT/enhancer binding protein(C/EBP)b,C/EBP a and peroxisome proliferator-activated receptor g(PPAR g)and PPAR target genes,including adipocyte fatty acid binding protein(aP2)and fatty acid synthase(FAS)were investigated.Results:ISA promoted 3T3-L1 fibroblast differentiation to adipocytes and increased triglyceride content by 26%.On mechanistic levels,ISA increased expressions of C/EBP b,PPAR g,C/EBP a,aP2 and FAS.Moreover,ISA stimulated expressions of adipokines secreted by adipocytes,including adiponectin,leptin,and resistin.Conclusions:These findings demonstrated that ISA promoted adipogenesis by up-regulating expressions of C/EBP b,PPAR g,C/EBP a,aP2 and FAS,and also stimulated adipokines during adipocyte differentiation.Further study should clarify the relationship between stimulation of adipokines and cognitive enhancing effect of ISA.展开更多
Mangoes (Mangifera indica L.) are one of the most important tropical foods. The seed is one of the main by-products of mango processing. Therefore, it is important to find an economically viable use for this waste (e....Mangoes (Mangifera indica L.) are one of the most important tropical foods. The seed is one of the main by-products of mango processing. Therefore, it is important to find an economically viable use for this waste (e.g., as a food additive or supplement with high nutraceutical value). We investigated the anti-obesity effects of mango seed kernel extract with hot water (MSKE-W) in 3T3-L1 adipocytes and in a high fat diet (HFD)-induced obesity rat model. MSKE-W caused a significant decrease in the activity of glycerol 2-phosphate dehydrogenase in 3T3-L1 adipocytes without eliciting cell cytotoxicity and inhibited cellular lipid accumulation through down-regulation of transcription factors such as PPARγ and C/EBPα. In the animal model, rats fed an HFD containing 1% MSKE-W gained less weight than rats fed an HFD alone. The visceral fat mass in rats fed an HFD containing 1% MSKE-W tended to be lower than that in rats fed an HFD alone. Furthermore, histological examination of rat livers from an HFD showed steatohepatitis. However, rats on an HFD containning 1% MSKE-W showed no histopathological changes in liver tissue. Our results indicate that MSKE-W influences anti-obesity effects, both in vitro and in vivo, and suggest that MSKE-W provides a novel preventive potential against obesity.展开更多
6a-Hydroxylup-20(29)-en-3-on-28-oic acid(1),a natural triterpenoid,was found to possess the ability in a dose-dependent manner inhibiting hormone-induced adipocyte differentiation in 3T3-L1 preadipocytes,and restoring...6a-Hydroxylup-20(29)-en-3-on-28-oic acid(1),a natural triterpenoid,was found to possess the ability in a dose-dependent manner inhibiting hormone-induced adipocyte differentiation in 3T3-L1 preadipocytes,and restoring glucose consuming ability in dexamethasone(DXM)-induced insulin resistant 3T3-L1 adipocytes.Compound 1 was also found to ameliorate DXM-induced adipocyte dysfunction in lipolysis and adipokine secretion.Mechanistic studies revealed that 1 inhibited adipocyte differentiation in 3T3-L1 preadipocytes via down-regulating hormone-stimulated gene transcription of peroxisome proliferator-activated receptor c and CCAAT-enhancer-binding protein alpha which are key factors in lipogenesis,and restored DXM-impaired glucose consuming ability in differentiated 3T3-L1 adipocytes via repairing insulin signaling pathway and activating down-stream signaling transduction by phosphorylation of signaling molecules PI3K/p85,Akt2 and AS160,thus leading to increased translocation of glucose transporter type 4 and transportation of glucose.展开更多
Summary: In order to investigate the effect of tumor necrosis factor-α (TNFα) on resistin expression in 3T3-L1 adipocytes, and further explore its mechanisms, the differentiated 3T3-L1 adipocytes were incubated with...Summary: In order to investigate the effect of tumor necrosis factor-α (TNFα) on resistin expression in 3T3-L1 adipocytes, and further explore its mechanisms, the differentiated 3T3-L1 adipocytes were incubated with 0, 1, 10, 100 ng/mL TNFα respectively for 24 h, and then the expression of resistin was determined. The differentiated 3T3-L1 adipocytes were incubated with 100 ng/mL TNFα for 3, 6, 24 h respectively, and then the expression of resistin mRNA was analyzed. 3T3-L1 adipocytes were induced to differentiate into mature adipocytes. The cells were randomly divided into 4 groups for culture. In the control group, no drugs were added. Cells of TNFα group were treated with 100 ng/mL TNFα. In Ro-31-8220 group, 5 μmol/L protein kinase C inhibitor Ro-31-8220 was added. With TNFα+Ro-31-8220 group, 100 ng/mL TNFα were added 1 h after the addition of 5 μmol/L Ro-31-8220. All adipocytes were cultured for 24 h. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were employed to detect the expression of resistin gene. Our results showed that resistin protein and mRNA in 3T3-L1 adipocytes were inhibited by TNFα at different concentrations (P<0.01), and the inhibitory effect increased with the concentration (P<0.01). At the same concentrations, the inhibitory effect increased with time (P<0.01). Ro-31-8220 could inhibit its expression and the inhibitive effect remained unchanged with addition of TNFα(P>0.05). It was concluded that TNFα could inhibit the expression of resistin in 3T3-L1 adipocytes. The mechanism may be that the expression of resistin is partly controlled by protein kinase C signal conduction pathway.展开更多
The anti-adipogenic and anti-obesity activity of chloroform fraction of Tephrosia purpurea (CFTp) on 3T3-L1 adipocytes and high fat diet (HFD)-fed obese rats was evaluated in this study. A substantial and dose depende...The anti-adipogenic and anti-obesity activity of chloroform fraction of Tephrosia purpurea (CFTp) on 3T3-L1 adipocytes and high fat diet (HFD)-fed obese rats was evaluated in this study. A substantial and dose dependent inhibition of α-glucosidase (81%) and lipase (75%) activities by CFTp was noticed. Treatment with CFTp (250 μg/mL) significantly inhibited 3T3-L1 adipocytes differentiation and lipid accumulation. A semi-quantitative RT-PCR analysis of 3T3-L1 cells revealed down regulation of mRNA expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS) and acetyl CoA carboxylase-2 (ACC-2), while glucose transporter type-4 (GLUT-4) expression was up-regulated in a dose dependent manner with CFTp. Further, oral administration of CFTp (200 mg/kg.b.wt.) significantly reduced body weight gain, fat mass, blood glucose and leptin levels in high fat diet (HFD)-induced obese rats. Taken together, these findings demonstrate that CFTp possesses potent anti-obesity activities.展开更多
Background: Adipocyte and osteoblast derive from the same mesenchimal progenitor. Age-related decrease in bone mass is accompanied by an increase in marrow adipose tissue. Vitamin D3 (VD3) inhibits adipogenesis in 3T3...Background: Adipocyte and osteoblast derive from the same mesenchimal progenitor. Age-related decrease in bone mass is accompanied by an increase in marrow adipose tissue. Vitamin D3 (VD3) inhibits adipogenesis in 3T3-L1 preadipocytes. Recently it has been demonstrated that alendronate (ALN) inhibits adipogenesis while promoting osteoblast differentiation of mesenchimal stem cells. Aim of the Study: To evaluate the role of ALN on adipocyte differentiation in vitro and the potential synergic role of VD3 co-treatment. Procedures: Murine 3T3-L1 and 3T3-F442A preadipocytes were routinely differentiated in presence of ALN and VD3 10-9 - 10-7 M for 7 days and then stained with Oil Red O. The effect of these treatments on mRNA expression of the main molecular markers of adipocyte differentiation (PPARγ and C/EBPα) and VD Receptor (VDR) were analyzed through RT-PCR. Results: Both ALN and VD3 showed a marked anti-adipogenic effect on 3T3-L1 cells. Co-incubation of ALN 10-8 M and VD3 10-9 M displayed no synergic effect on inhibition of adipogenesis. PPARγ mRNA expression was significantly reduced by ALN and VD3. mRNA expression of C/EBPα was reduced only by VD3 treatment. An increase in VDR mRNA expression of 3T3-L1 cells was observed with both ALN and VD3. On the contrary, 3T3-F442A cells, which are in a more advanced adipogenic differentiation stage compared to 3T3-L1, did not express detectable levels of VDR. Interestingly, adipose differentiation of 3T3-F442A was not affected by ALN nor VD3. These results suggest that VDR may represent the molecular target of the anti-adipogenic effect of ALN. Conclusion: VDR plays a critical role in mediating the anti-adipogenic effect of ALN. Further studies to clarify this mechanism are warranted.展开更多
基金This work was graciously supported by the Chinese National Natural Science Foundation(Grant 31901725 and 32201933)the Science and Technology Projects in Guangzhou(Grant 202201010170).
文摘Vaccarin,a flavonoid glycoside isolated from Vaccaria segetalis,is non-toxic to 3T3-L1 cells up to concentrations of 200μM.Accordingly,we investigated the effects of this natural product on adipogenesis and lipolysis in 3T3-L1 adipocytes.Our results revealed that vaccarin significantly inhibited lipid accumulation by suppressing the adipogenesis-related transcription factors peroxisome proliferator-activated receptorγ(PPARγ)and the CCAAT/enhancer-binding proteinα(C/EBPα).Specifically,lipid accumulation decreased by up to 27.7±2.7%when 3T3-L1 adipocytes were treated with a 10μM concentration of vaccarin.Mechanistic studies showed that the compound inhibited adipogenesis through activation of the Hedgehog(Hh)signaling pathway and so restoring Smo and Gli1 expression at an early stage of differentiation.In mature 3T3-L1 cells,vaccarin significantly increased the secretion of glycerol into the surrounding medium and thus indicating that it accelerated the degradation of triglycerides.In addition,vaccarin,was shown to enhance lipolysis through stimulation of the transcription levels of lipoprotein lipase,monoglycerides lipase,adipose triacylglyceride lipase,hormone-sensitive lipase and adipose differentiated-related protein.All told,vaccarin suppressed lipid accumulation and enhanced lipolysis during adipocyte differentiation by restoring Hh signaling.As such,it is a phytochemical capable of halting adipocyte hyperplasia and,thereby,ameliorating the effects of obesity.
基金This work was supported by National Natural Science Foundation of China(31900292)Key Project in Science and Technology Agency of Henan Province(182102410083).
文摘Ligustri lucidi Fructus is a traditional Chinese medicine and possesses various bioactivities,including hypoglycemic effect.Ligustrum lucidum Ait flowers are poorly investigated.Thus,we hypothesized that L.lucidum flowers also could have hypoglycemic effect.Chemical composition and glucose uptake effect of L.lucidum flowers on 3T3-L1 adipocytes were investigated.In this study,the components of L.lucidum flowers were investigated by various chromatographic and spectroscopic methods and the effects of L.lucidum flowers on the induction of glucose uptake were investigated by 3T3-L1 adipocytes.Seven compounds were isolated and identified from L.lucidum flowers,including ursolic acid(1),kaempferol-7-O-α-Lrhamnoside(2,KR),β-sitosterol(3),β-daucosterol(4),kaempferitrin(5,KF),10-hydroxy oleuropein(6,HO),and kaempferol-3-O-β-D-glucopyranoside-7-α-L-rhamnopyranoside(7,KGR).The results of glucose uptake showed that total extract(TE),KR,KF and HO from L.lucidum flowers significantly enhanced glucose uptake.This study demonstrated that compounds 2,5-7 were isolated for the first time from this plant and compounds 2 and 7 were isolated for the first time in genus Ligustrum,L.lucidum flowers,KR,KF and HO may possess potential hypoglycemic effect.
基金supported by the National 863 Program of China (2006AA10Z332)the Ear Marked Fund for Modern Agro-Industry Technology Research System, China
文摘The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food to treat obesity and diabetes, The isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography. With berberine as a positive control, different concentrations ofsporamin （0.000, 0.125, 0.025, 0.250, 0.500, and 1.000 mg·mL^-1 were used to treat 3T3-L1 preadipocytes. Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry, Preadipocytes differentiation was measured by 3（4,5-dimethylthiazolyl-2-yl）-2,5-diphenyltetrazolium bromide （MTT） spectrophotometric assay. Two sporamin proteins, which were separated into sporamin A （31 kD） and sporamin B （22 kD）, could be purified by ion-exchange and gel filtration chromatography. After being treated by different concentrations of sporamin, the differentiation of 3T3-L1 preadipocytes was significantly inhibited, compared with the positive control. When the sporamin solution concentration was 0.500 mg mL-1, the accumulation of lipid droplets within the cells was significantly decreased and the optical density （OD） value of the solution from destained Oil Red O reached to 0.35, which was the lowest value （P〈 0.05）. The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations. In addition, the inhibitory effect was more obvious with the prolonged treatment time （P〈 0.05）. The differentiation and proliferation of 3T3-L1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin. It was, therefore, suggested that the sporamin was potentially effective for weight loss.
文摘Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained with OilRed-O staining to confirm adipogenesis.The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay,intracellular reactive oxygen species generation through flow cytometry,and morphologically through confocal microscopy using propidium iodide,4,6-diamino-2-phenylindol dihydrochloride,and 2’,7’-dichlorofluorescein diacetate dyes.The uptake of fluorescent glucose analog,2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy.Results:MTT assay showed that the cell survival rate was(28.00±3.00)%,(92.33±2.60)%,and(71.34±2.10)%in the presence of 2 mM H2O2,AbPI alone,and AbPI and H2O2 both,respectively,in comparison to the control.Oil-Red-O staining indicated that Ab PI enhanced adipogenesis.AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone,and showed insulinsensitizing effect in the presence of insulin,but failed to stimulate the uptake in the absence of insulin.Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon Ab PI treatment.Confocal microscopy showed that the damaged cell population rose to 3.50%,117.84%,and 261.50%in the presence of Ab PI alone,AbPI with H2O2,and H2O2 alone,respectively.Conclusions:The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.
基金Mukogawa Women’s University Short-term Student Exchange Program,Science Foundation for The Excellent Youth Scholars of Beijing University of Chinese Medicine(No.2012-QNJSZX006)Natural Science Foundation of Beijing Municipality(No.7144222).
文摘Objective:3,4-Oxo-isopropylidene-shikimic acid(ISA),a derivative of shikimic acid,has exhibited ameliorative effect on cognitive impairment in experimental animal models of dementia.This study investigated the effect of ISA on lipid accumulation and adipokine secretion during differentiation of 3T3-L1 fibroblasts to adipocytes.Methods:3T3-L1 cells were cultured and treated with ISA(50e800 mM)from days 3e8.Lipid accumulation and triglyceride content were measured.Gene expression of adipokines(adiponectin,leptin,and resistin),CCAAT/enhancer binding protein(C/EBP)b,C/EBP a and peroxisome proliferator-activated receptor g(PPAR g)and PPAR target genes,including adipocyte fatty acid binding protein(aP2)and fatty acid synthase(FAS)were investigated.Results:ISA promoted 3T3-L1 fibroblast differentiation to adipocytes and increased triglyceride content by 26%.On mechanistic levels,ISA increased expressions of C/EBP b,PPAR g,C/EBP a,aP2 and FAS.Moreover,ISA stimulated expressions of adipokines secreted by adipocytes,including adiponectin,leptin,and resistin.Conclusions:These findings demonstrated that ISA promoted adipogenesis by up-regulating expressions of C/EBP b,PPAR g,C/EBP a,aP2 and FAS,and also stimulated adipokines during adipocyte differentiation.Further study should clarify the relationship between stimulation of adipokines and cognitive enhancing effect of ISA.
文摘Mangoes (Mangifera indica L.) are one of the most important tropical foods. The seed is one of the main by-products of mango processing. Therefore, it is important to find an economically viable use for this waste (e.g., as a food additive or supplement with high nutraceutical value). We investigated the anti-obesity effects of mango seed kernel extract with hot water (MSKE-W) in 3T3-L1 adipocytes and in a high fat diet (HFD)-induced obesity rat model. MSKE-W caused a significant decrease in the activity of glycerol 2-phosphate dehydrogenase in 3T3-L1 adipocytes without eliciting cell cytotoxicity and inhibited cellular lipid accumulation through down-regulation of transcription factors such as PPARγ and C/EBPα. In the animal model, rats fed an HFD containing 1% MSKE-W gained less weight than rats fed an HFD alone. The visceral fat mass in rats fed an HFD containing 1% MSKE-W tended to be lower than that in rats fed an HFD alone. Furthermore, histological examination of rat livers from an HFD showed steatohepatitis. However, rats on an HFD containning 1% MSKE-W showed no histopathological changes in liver tissue. Our results indicate that MSKE-W influences anti-obesity effects, both in vitro and in vivo, and suggest that MSKE-W provides a novel preventive potential against obesity.
基金China National Major Projects of Science&Technology(2014ZX100050022009ZX09103-436)+1 种基金the Young Academic Leader Raising Foundation of Yunnan Province(No.2009CI073)the foundation from Chinese Academy of Sciences to Gang Xu,and the Program for Research Team in South China Chinese Medicine Collaborative Innovation Center of Guangdong,China(A1-AFD01514A07).
文摘6a-Hydroxylup-20(29)-en-3-on-28-oic acid(1),a natural triterpenoid,was found to possess the ability in a dose-dependent manner inhibiting hormone-induced adipocyte differentiation in 3T3-L1 preadipocytes,and restoring glucose consuming ability in dexamethasone(DXM)-induced insulin resistant 3T3-L1 adipocytes.Compound 1 was also found to ameliorate DXM-induced adipocyte dysfunction in lipolysis and adipokine secretion.Mechanistic studies revealed that 1 inhibited adipocyte differentiation in 3T3-L1 preadipocytes via down-regulating hormone-stimulated gene transcription of peroxisome proliferator-activated receptor c and CCAAT-enhancer-binding protein alpha which are key factors in lipogenesis,and restored DXM-impaired glucose consuming ability in differentiated 3T3-L1 adipocytes via repairing insulin signaling pathway and activating down-stream signaling transduction by phosphorylation of signaling molecules PI3K/p85,Akt2 and AS160,thus leading to increased translocation of glucose transporter type 4 and transportation of glucose.
文摘Summary: In order to investigate the effect of tumor necrosis factor-α (TNFα) on resistin expression in 3T3-L1 adipocytes, and further explore its mechanisms, the differentiated 3T3-L1 adipocytes were incubated with 0, 1, 10, 100 ng/mL TNFα respectively for 24 h, and then the expression of resistin was determined. The differentiated 3T3-L1 adipocytes were incubated with 100 ng/mL TNFα for 3, 6, 24 h respectively, and then the expression of resistin mRNA was analyzed. 3T3-L1 adipocytes were induced to differentiate into mature adipocytes. The cells were randomly divided into 4 groups for culture. In the control group, no drugs were added. Cells of TNFα group were treated with 100 ng/mL TNFα. In Ro-31-8220 group, 5 μmol/L protein kinase C inhibitor Ro-31-8220 was added. With TNFα+Ro-31-8220 group, 100 ng/mL TNFα were added 1 h after the addition of 5 μmol/L Ro-31-8220. All adipocytes were cultured for 24 h. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were employed to detect the expression of resistin gene. Our results showed that resistin protein and mRNA in 3T3-L1 adipocytes were inhibited by TNFα at different concentrations (P<0.01), and the inhibitory effect increased with the concentration (P<0.01). At the same concentrations, the inhibitory effect increased with time (P<0.01). Ro-31-8220 could inhibit its expression and the inhibitive effect remained unchanged with addition of TNFα(P>0.05). It was concluded that TNFα could inhibit the expression of resistin in 3T3-L1 adipocytes. The mechanism may be that the expression of resistin is partly controlled by protein kinase C signal conduction pathway.
文摘The anti-adipogenic and anti-obesity activity of chloroform fraction of Tephrosia purpurea (CFTp) on 3T3-L1 adipocytes and high fat diet (HFD)-fed obese rats was evaluated in this study. A substantial and dose dependent inhibition of α-glucosidase (81%) and lipase (75%) activities by CFTp was noticed. Treatment with CFTp (250 μg/mL) significantly inhibited 3T3-L1 adipocytes differentiation and lipid accumulation. A semi-quantitative RT-PCR analysis of 3T3-L1 cells revealed down regulation of mRNA expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS) and acetyl CoA carboxylase-2 (ACC-2), while glucose transporter type-4 (GLUT-4) expression was up-regulated in a dose dependent manner with CFTp. Further, oral administration of CFTp (200 mg/kg.b.wt.) significantly reduced body weight gain, fat mass, blood glucose and leptin levels in high fat diet (HFD)-induced obese rats. Taken together, these findings demonstrate that CFTp possesses potent anti-obesity activities.
文摘Background: Adipocyte and osteoblast derive from the same mesenchimal progenitor. Age-related decrease in bone mass is accompanied by an increase in marrow adipose tissue. Vitamin D3 (VD3) inhibits adipogenesis in 3T3-L1 preadipocytes. Recently it has been demonstrated that alendronate (ALN) inhibits adipogenesis while promoting osteoblast differentiation of mesenchimal stem cells. Aim of the Study: To evaluate the role of ALN on adipocyte differentiation in vitro and the potential synergic role of VD3 co-treatment. Procedures: Murine 3T3-L1 and 3T3-F442A preadipocytes were routinely differentiated in presence of ALN and VD3 10-9 - 10-7 M for 7 days and then stained with Oil Red O. The effect of these treatments on mRNA expression of the main molecular markers of adipocyte differentiation (PPARγ and C/EBPα) and VD Receptor (VDR) were analyzed through RT-PCR. Results: Both ALN and VD3 showed a marked anti-adipogenic effect on 3T3-L1 cells. Co-incubation of ALN 10-8 M and VD3 10-9 M displayed no synergic effect on inhibition of adipogenesis. PPARγ mRNA expression was significantly reduced by ALN and VD3. mRNA expression of C/EBPα was reduced only by VD3 treatment. An increase in VDR mRNA expression of 3T3-L1 cells was observed with both ALN and VD3. On the contrary, 3T3-F442A cells, which are in a more advanced adipogenic differentiation stage compared to 3T3-L1, did not express detectable levels of VDR. Interestingly, adipose differentiation of 3T3-F442A was not affected by ALN nor VD3. These results suggest that VDR may represent the molecular target of the anti-adipogenic effect of ALN. Conclusion: VDR plays a critical role in mediating the anti-adipogenic effect of ALN. Further studies to clarify this mechanism are warranted.
