染料木黄酮抑制3T3-L1细胞成脂分化机制  

Anti-adipogenic Effect of Genistein in 3T3-L1 Cells and Its Mechanism

作　　者：胡文敏[1] 张岭[1] 李林子[1] 张丽婧[1] 胡志航[1] 陈建国[1] 刘冬英[1] 刘臻[1] 王茵[1] 

机构地区：[1]浙江省医学科学院,浙江杭州310013

出　　处：《食品科学》2016年第17期219-224,共6页Food Science

基　　金：浙江省医药卫生平台重点资助计划项目(2014ZDA004);浙江省营养学医学支撑学科建设项目(11-zc03);浙江省科技厅创新团队建设与人才培养项目(2011F20038);浙江省科技厅支撑学科建设项目(2013F10007);浙江省151人才培养项目;食品(保健食品)质量安全评价研究实验室建设项目(2015F10014)

摘　　要：目的：研究染料木黄酮是否通过雌激素受体介导影响3T3-L1前脂肪细胞成脂分化,并探讨其可能的机制。方法：以不同浓度染料木黄酮处理3T3-L1细胞,四甲基偶氮唑蓝（methyl thiazolyl tetrazolium,MTT）法测定细胞存活率;在3T3-L1前脂肪细胞成脂分化过程中分别加入不同浓度染料木黄酮、染料木黄酮和ICI182780（一种雌激素受体抑制剂）混合物及不同浓度雌二醇,油红染色观察分化结果,测定细胞甘油三酯（triglyceride,TG）含量;染料木黄酮、染料木黄酮和ICI182780混合物及不同浓度雌二醇处理诱导成熟后的脂肪细胞,测定培养液甘油含量;分别用染料木黄酮（30μmol/L）、染料木黄酮（30μmol/L）和ICI182780混合物干预细胞成脂分化,Western blotting法检测细胞外信号调节激酶（extracellular signal-regulated kinase,ERK）、p-ERK、腺苷酸活化蛋白激酶（adenosine monophosphate activated protein kinase,AMPK）、p-AMPK、激素敏感性脂肪酶（hormone sensitive lipase,HSL）、脂肪酸合成酶（fatty acid synthetase,FAS）蛋白表达量。结果：3T3-L1细胞存活率随染料木黄酮浓度的升高而降低,50-200μmol/L染料木黄酮能抑制细胞生长（P〈0.01）;染料木黄酮能负向调节成脂分化后细胞胞浆内TG含量,在0-50μmol/L浓度范围内呈剂量依赖关系,高剂量雌二醇（100 nmol/L）能下调TG含量;染料木黄酮能促进成熟脂肪细胞脂解,提高细胞培养液甘油浓度;染料木黄酮（30μmol/L）能上调p-AMPK、HSL蛋白表达,下调FAS蛋白表达（P〈0.01）,对ERK、p-ERK、AMPK表达量无明显影响（P〉0.05）。ICI182780（1μmol/L）能部分阻断染料木黄酮（30μmol/L）对p-AMPK的调节作用（P〈0.05）。结论：染料木黄酮能抑制3T3-L1细胞成脂分化,其机制可能是染料木黄酮一方面下调FAS蛋白表达,抑制脂肪合成,另一方面激活AMPK-HSL途径促进脂肪分解;染料木黄酮还可能通过非雌激素受体途径Objective： To investigate the anti-adipogenic effect and underlying mechanism of genistein in 3T3-L1 cells and its estrogen receptor（ER） mediated pathway. Methods： After treated with various concentrations of genistein, the survival rate of 3T3-L1 cells was assessed by MTT assay. In addition, after treated with genistein, genistein-ICI182780, or estrogen, differentiation degree and lipid accumulation of the cells were assessed by oil red staining and a TG assay kit, and adipocyte lipolysis was assessed by a glycerin assay kit. After intervention by genistein or genistein-ICI182780, the expression of extracellular signal-regulated kinase（ERK）, phospho-extracellular signal-regulated kinase（p-ERK）, adenosine monophosphate activated protein kinase（AMPK）, phospho-adenosine monophosphate activated protein kinase（p-AMPK）, hormone sensitive lipase（HSL）, and fatty acid synthetase（FAS） were analyzed by Western blotting. Results： Genistein and high-dose estrogen inhibited lipid accumulation of 3T3-L1 and increased lipolysis in adipocytes. The presence of ICI182780 partly decreased both functions of genistein. Furthermore, genistein increased the expression of p-AMPK and HSL, and decreased the expression of FAS. The presence of ICI182780 could slightly suppress the increase of p-AMPK caused by genistein intervention. Conclusion： Genistein can inhibit the adipogenesis and differentiation of 3T3-L1 cells via a non-ER pathway through a mechanism which may be relate to p-AMPK, HSL and FAS.

关 键 词：染料木黄酮 3T3-L1细胞 磷酸化腺苷酸活化蛋白激酶 激素敏感性脂肪酶 脂肪酸合成酶 

分 类 号：R151[医药卫生—营养与食品卫生学]