白藜芦醇对小鼠3T3-F442A脂肪细胞增殖与脂肪生成的影响

Effects of Resveratrol on Cell Proliferation and Adipogenesis in Murine 3T3-F442A Preadipocytes

研究白藜芦醇(Resveratrol,Res)对小鼠3T3-F442A脂肪细胞增殖与脂肪生成的影响。采用MTT法检测不同剂量的Res在24、48、72 h对3T3-F442A前脂肪细胞增殖的影响,得到72 h的半抑制浓度并确定给药浓度;使用添加40μmol/L Res的诱导分化培养基诱导3T3-F442A前脂肪细胞分化为成熟脂肪细胞,通过油红O染色法观察Res对3T3-F442A脂肪细胞脂肪生成的影响;采用实时荧光定量PCR技术、蛋白免疫印迹实验、酶联免疫吸附实验检测Res对3T3-F442A脂肪细胞分化相关转录因子(C/EBPα、C/EBPβ、PPARγ和aP2)、脂肪生成相关血管内皮生长因子及受体(VEGF-α和VEGFR-2)以及基质金属蛋白酶(MMP-2和MMP-9)表达的影响。Res以时间-剂量依赖方式抑制3T3-F442A前脂肪细胞增殖;Res组的脂肪细胞分化相关转录因子、血管重塑因子以及基质金属蛋白酶(MMP-2和MMP-9)mRNA表达水平均明显下调;脂肪细胞分化相关转录因子的蛋白表达水平均明显下调以及分泌到胞外的基质金属蛋白酶(MMP-2和MMP-9)的浓度也明显下降。Res能抑制3T3-F442A前脂肪细胞增殖与脂肪生成,其机制可能是通过抑制与基质金属蛋白酶(MMP-2和MMP-9)相关的血管生成过程而发生的。

The effects of Resveratrol(Res)on the proliferation and adipogenesis of 3T3-F442A adipocytes in murine mice were investigated.The effects of different doses of Res on the proliferation of 3T3-F442A preadipocytes at 24,48 and 72 h were detected by MTT assay,and the semi-inhibitory concentration at 72 h was obtained and determined the optimal concentration;and then 3T3-F442A preadipocytes were induced to differentiate into mature adipocytes by using the induced differentiation medium with the addition of 40μmol/L Res,and the effects of Res on the adipogenesis of 3T3-F442A adipocytes were observed by the oil-red O staining method.Quantitative Real-time PCR,Western blot,and enzyme-linked immunosorbent assay were used to detect the effects of Res on 3T3-F442A adipocyte differentiation-associated transcription factors(C/EBPα,C/EBPβ,PPARγ,and aP2),adipogenesis-associated vascular endothelial growth factor and receptors(VEGF-αand VEGFR-2),and matrix metalloproteinases(MMP-2 and MMP-9)expression.Res inhibited the proliferation of 3T3-F442A preadipocytes in a time-dose-dependent manner;mRNA expression levels of adipocyte differentiation-associated transcription factors,vascular remodelling factors,and matrix metalloproteinases(MMP-2 and MMP-9)were significantly down-regulated in the Res group,and Protein expression levels of adipocyte differentiation-associated transcription factors were significantly down-regulated,as well as the levels of matrix metalloproteinases(MMP-2 and MMP-9)secreted into the extracellular compartment.Res inhibited the proliferation and adipogenesis of 3T3-F442A preadipocytes,probably by inhibiting the angiogenic process associated with matrix metalloproteinases(MMP-2 and MMP-9).