姜黄素通过减少CREB转录活性抑制前脂肪细胞分化

摘要

Aim To investigate the effect of curcumin on differentiation of 3T3-L1 preadipocytes and its mechanism.Methods 3T3-L1 preadipocytes were treated with curcumin at different concentrations (10,20,40 μmol · L-1) and cell differentiation was synergistically induced.The proliferation of 3T3-L1 preadipocytes was detected by MTT assay.The lipid accumulation was detected by oil red O staining method,and the levels of CREB,C/EBPβ,C/EBPα,PPARγmRNA and protein in 3T3-L1 preadipocytes were detected by qRT-PCR and Western blot.Results MTT results showed that curcumin had a significant inhibitory effect on proliferation of 3T3-L1 preadipocytes in a dose-and time-dependent manner.Oil red O staining showed that curcumin inhibited the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner.When 0,20,40 μmol · L-1 curcumin treatment of 3T3-L1 preadipocytes were performed,the levels of p-CREB,C/EBPβ,C/EBPα,PPARγ mRNA and protein significantly decreased with the increasing dose of curcumin (P ＜ 0.05).Conclusions Curcumin can effectively inhibit the proliferation and differentiation of 3T3-L1 preadipocytes.The mechanism may be that curcumin inhibits CREB transcriptional activity and inhibits the expression of C/EBPβ,C/EBPα and PPARγ.%目的 探讨姜黄素对3T3-L1前脂肪细胞分化的影响及其作用机制.方法 选取3T3-L1前脂肪细胞为研究对象,姜黄素(10、20、40 μmol·L-1)处理3T3-L1前脂肪细胞并同步诱导细胞分化,采用MTT法检测3T3-L1前脂肪细胞增殖情况,油红O染色方法检测胞质脂质堆积情况,实时荧光定量PCR法及蛋白印迹法检测3T3-L1脂肪细胞环磷腺苷效应元件结合蛋白(CREB)、CCAAT/增强子结合蛋白β(C/EBPβ)、CCAAT/增强子结合蛋白α(C/EBPα)、过氧化物体增殖剂活化受体γ (PPARγ)的mRNA及蛋白表达水平.结果 MTT结果显示,姜黄素对3T3-L1前脂肪细胞增殖具有明显抑制作用,且呈现一定剂量、时间依赖性;油红0染色显示,姜黄素可抑制3T3-L1前脂肪细胞分化,且呈现一定剂量依赖性;0、20、40 μmol·L-1姜黄素处理3T3-L1前脂肪细胞,p-CREB、C/EBPβ、C/EBPα、PPARγ mRNA水平及蛋白水平随姜黄素剂量升高均呈现明显下降趋势(P＜0.05).结论 姜黄素可有效抑制3T3-L1前脂肪细胞增殖分化,其机制可能为姜黄素抑制CREB转录活性,从而抑制C/EBPβ、C/EBPα、PPARγ表达.