硫酸脱氢表雄酮抑制3T3-L1前脂肪细胞分化的分子机制研究  

Effects of dehydroepiandrosterone sulfate on 3T3-L1 pre-adipocyte differentiation and its potential mechanism

作　　者：瞿茹怡 徐玉梅 蒋英 郑洁[2] QU Ruyi;XU Yumei;JIANG Ying;ZHENG Jie(Department of Molecular Diagnosis,Affiliated Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Department of Molecular Diagnosis,Shanghai 200011,China;Department of Geriatrics,Shanghai Fourth People’s Hospital)

机构地区：[1]上海交通大学医学院附属第九人民医院分子诊断科,上海200011 [2]上海市第四人民医院老年医学科

出　　处：《山西医科大学学报》2019年第8期1116-1121,共6页Journal of Shanxi Medical University

基　　金：上海市卫计委资助面上项目(201440446)

摘　　要：目的探讨硫酸脱氢表雄酮(DHEA-S)对3T3-L1前脂肪细胞株分化的影响及可能的分子机制。方法以3T3-L1细胞株为研究对象,先观察不同时间点不同剂量的DHEA-S对3T3-L1细胞活力的影响,确定合适的干预浓度。3T3-L1细胞分为DMSO组(对照组)和DHEA-S组,观察DHEA-S对3T3-L1前脂肪细胞分化的影响。通过测定甘油醛3-磷酸脱氢酶(G3PDH)活性和用油红O染色计算的脂肪细胞分化率,评估脂肪细胞分化的变化;用real time-PCR法观察诱导前(第0天)和诱导后不同时间点(第1,2,4,6,10,14天)目的基因过氧化物酶体增殖物激活受体-γ(PPARγ)、固醇调节元件结合蛋白-1c(SREBP1-c)、CCAAT增强子结合蛋白家族(C/EBPs)、11β-羟类固醇脱氢酶1(11HSDB1)、激素敏感性脂肪酶(HSL)和脂蛋白酯酶(LPL)mRNA表达的变化。结果MTT实验结果显示DHEA-S在50μmol/L时对细胞活力影响<30%,对后续的干预影响较小。分化后第4天对照组和DHEA-S组的G3PDH活性分别为(380.83±28.43)nmol/(min·mg)和(253.07±27.67)nmol/(min·mg),分化后第10天对照组和DHEA-S组的G3PDH活性为(573.93±17.5)nmol/(min·mg)和(398.17±24.05)nmol/(min·mg),差异均具有统计学意义(P<0.01)。油红O染色计算脂肪细胞分化率对照组和DHEA-S组在诱导后第4天分别为39.09%±5.51%和15.66%±0.75%,第10天分别为78.82%±3.37%和49.09%±9.75%,差异也有统计学意义(P<0.01)。real time PCR检测结果显示,DHEA-S显著抑制脂肪细胞分化相关的关键基因(PPARγ、ADD1/SREBP1-c、C/EBPs、11HSDB1、LPL和HSL)mRNA的表达。结论DHEA-S(50μmol/L)在体外可能通过抑制11HSDB1的表达,减少局部活性皮质醇,进而下调其他脂肪细胞分化的关键基因,抑制3T3-L1细胞的分化。Objective To investigate the effects of dehydroepiandrosterone sulfate(DHEAS)on 3T3-L1 pre-adipocyte differentiation and its possible mechanism.Methods The optimal DHEAS concentration was determined based on cell viability measured by MTT.3T3-L1 cells were divided into DMSO group and DHEA-S group.The change of adipocyte differentiation was assessed by measuring G3PDH activity and calculating adipocyte differentiation rate by oil red O staining after 50μmol/L DHEA-S treatment.Real time-PCR was used to observe the mRNA expression of the target genes,peroxisome proliferator-activated receptor-gamma(PPARγ),sterol regulatory element binding protein-1c(SREBP1-c),CCAAT enhancer binding protein family(C/EBPs),11β-hydroxysteroid dehydrogenase 1(11HSDB1),hormone sensitive lipase(HSL)and lipoprotein lipase(LPL)before induction(d0)and at different time points after induction(d1,d2,d4,d6,d10 and d14).Results DHEA-S had little effect(<30%)on cell viability at 50μmol/L by MTT.The G3PDH activity in DHEA-S group was significantly lower than that in DMSO group at day 4[(380.83±28.43)nmol/(min·mg)vs(253.07±27.67)nmol/(min·mg),P<0.01]and day 10 after differentiation[(573.93±17.5)nmol/(min·mg)vs(398.17±24.05)nmol/(min·mg),P<0.01].The adipocyte differentiation rate in DHEA-S group was significantly lower than that in DMSO group at day 4(15.66%±0.75%vs 39.09%±5.51%,P<0.01)and day 10 after differentiation(49.09%±9.75%vs 78.82%±3.37%,P<0.01).Real-time PCR showed that DHEAS significantly inhibited the expression of key genes involved in adipocyte differentiation(PPARγ,ADD1/SREBP1-c,C/EBPs,11HSDB1,LPL and HSL).Conclusion DHEAS(50μmol/L)may inhibit the expression of 11HSDB1,reduce local active cortisol,further down-regulate the key genes regulating adipocyte differentiation,and then inhibit the differentiation of 3T3-L1 cells in vitro.

关 键 词：硫酸脱氢表雄酮 3T3-L1前脂肪细胞 脂肪细胞分化 CCAAT增强子结合蛋白 11β-羟类固醇脱氢酶1 

分 类 号：R363[医药卫生—病理学]