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当前位置: 首页 > 文章 > 染料木黄酮抑制3T3-L1细胞成脂分化机制 食品科学 2016,37 (17) 219-224
Position: Home > Articles > Anti-adipogenic Effect of Genistein in 3T3-L1 Cells and Its Mechanism FOOD SCIENCE 2016,37 (17) 219-224

染料木黄酮抑制3T3-L1细胞成脂分化机制

作  者：
胡文敏;张岭;李林子;张丽婧;胡志航;陈建国;刘冬英;刘臻;王茵
单  位：
浙江省医学科学院
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
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抑制3T3-L1细胞成脂分化。
译  名：
Anti-adipogenic Effect of Genistein in 3T3-L1 Cells and Its Mechanism
作  者：
HU Wenmin;ZHANG Ling;LI Linzi;ZHANG Lijing;HU Zhihang;CHEN Jianguo;LIU Dongying;LIU Zhen;WANG Yin;Zhejiang Academy of Medical Sciences;
关键词：
genistein;;3T3-L1 cell;;phospho-adenosine monophosphate activated protein kinase(p-AMPK);;hormone sensitive lipase(HSL);;fatty acid synthetase(FAS)
摘  要：
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.

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