【摘要】 背景:目前关于脂肪细胞分化的分子作用机制的研究较少。过氧化物酶增殖物激活受体和CCAAT/增强子结合蛋白家族的转录调控因子可诱导前脂肪细胞表达促进其分化成熟的多个转录因子,但其作用机制却罕见报道。目的:观察小鼠3T3-L1前脂肪细胞分化为成熟脂肪细胞过程中过氧化物酶增殖物激活受体γ和CCAAT/增强子结合蛋白α的表达,以及在脂肪细胞分化过程中的变化。方法:体外培养3T3-L1前脂肪细胞,采用经典激素鸡尾酒诱导法诱导细胞分化,诱导剂为1-甲基-3-异丁基-黄嘌呤、地塞米松和胰岛素。诱导后0,2,4,6和8d,用油红O染色和染色比色法分析脂肪细胞的分化程度,采用实时PCR和Western blot技术检测不同时间点的过氧化物酶增殖物激活受体γ和CCAAT/增强子结合蛋白α的表达。结果与结论:在前体脂肪细胞的分化过程中,随时间的延长相对脂肪含量、过氧化物酶增殖物激活受体γ与CCAAT/增强子结合蛋白α表达水平均明显升高(P<0.01)。说明在小鼠3T3-L1前脂肪细胞分化为成熟脂肪细胞过程中,过氧化物酶增殖物激活受体γ与CCAAT/增强子结合蛋白α对细胞分化可能起促进作用。更多还原

【Abstract】 BACKGROUND: Few studies were concerned about the molecular mechanisms in the process of adipocyte differentiation at present. The transcription factors of peroxisome proliferator-activated receptors (PPAR) and CCAAT/enhancer binding protein (C/EBP) family can induce and promote the differentiation, but the mechanisms were rarely reported. OBJECTIVE: To investigate PPAR γ and C/EBP α expression during the process of murine 3T3-L1 preadipocytes differentiating into mature adipocytes, and explore the mechanisms in the differentiation process of adipocytes. METHODS: Murine 3T3-L1 preadipocytes was cultured in vitro and induced by using the classic hormone-cocktail method (1-methyl-3-butyl-anthine + dexamethasone + insulin). At 0, 2, 4, 6 and 8 days following induction, oil red O staining method and spectrophotography were applied to analyze the differentiation of adipocytes. Real time polymerase chain reaction (PCR) and Western blot methods were applied to detect the expression of PPARγ and C/EBPα at different time points. RESULTS AND CONCLUSION: In the process of cell differentiation, the difference of relative fat content, expression levels of PPARγ and C/EBPα were significantly increased (P<0.01). These indicated that PPARγ and C/EBPα play a promoting effect in adipocyte differentiation from 3T3-L1 preadipocytes.更多还原