白芦藜醇影响前脂肪细胞增殖与分化及调控相关信号通路的研究

Effects of Resveratrol on Proliferation and Differentiation of Preadipocytes and Regulation of Related Signaling Pathways

目的：探讨白芦藜醇（Resveratrol）对于小鼠3T3-L1前脂肪细胞增殖与分化的影响及其调控相关信号通路的研究。方法：培养小鼠3T3-L1前脂肪细胞,添加不同剂量的白芦藜醇处理24、48 h,用MTT法检测细胞增殖;在诱导3T3-L1前脂肪细胞分化的过程中添加100μmol·L^（-1）白芦藜醇处理48 h,继续培养6 d后油红O染色观察脂肪细胞的分化程度;采用荧光定量PCR技术检测与分化相关基因过氧化物酶体增殖物激活受体γ（PPARγ）、CCAAT/增强子结合蛋白-α（C/EBPα）和Notch1,炎症相关基因单核细胞趋化因子-1（MCP-1）、肿瘤坏死因子-α（TNF-α）和瘦素（Leptin）的mRNA表达水平;酶联免疫吸附法（ELISA）分析3T3-L1分化过程中白芦藜醇对MCP-1蛋白分泌的影响。结果：白芦藜醇显著抑制3T3-L1前脂肪细胞增殖,并呈现一定的时间、剂量依赖效应。与对照组相比,白芦藜醇通过抑制脂肪细胞中分化相关基因（PPARγ、C/EBPα和Notch1）mRNAs表达水平,抑制3T3-L1细胞分化过程中脂滴的合成;白芦藜醇可以促进3T3-L1前脂肪细胞向脂肪细胞分化过程中炎症相关因子MCP-1的mRNA表达和蛋白分泌,白芦藜醇致成熟脂肪细胞中Leptin mRNA表达水平在第2天降低,但第4天升高。结论：白芦藜醇可以抑制3T3-L1前脂肪细胞增殖,及前脂肪细胞分化,其机制可能是通过抑制PPARγ、C/EBPα和Notch1的表达。在3T3-L1前脂肪细胞分化过程中,白芦藜醇可以促进脂肪细胞分化过程中MCP-1的表达及分泌,及增加Leptin基因表达,调控炎症状态影响肥胖。

Objective： To investigate the effect of resveratrol on the proliferation and differentiation of mice 3 T3-L1 preadipocytes and the regulation of related signaling pathways. Methods： Mice 3 T3-L1 preadipocytes were cultured and treated with different doses of resveratrol for 24 h and 48 h. Cell proliferation was detected by MTT assay. In the process of induction of 3 T3-L1 preadipocyte differentiation,100 μmol/L resveratrol was treated for 48 h,and the degree of differentiation of adipocytes was observed by oil red O staining after 6 days of continuous culture. PPARγ,CCAAT/enhancer binding protein-α（ C/EBPα） and Notch1,inflammation-related gene monocyte chemotactic factor（ MCP-1）,tumor necrosis factor-α（ TNF-α） and leptin（ leptin） were detected by fluorescence quantitative polymerase chain reaction（ RT-PCR）. Enzyme-linked immunosorbent assay（ ELISA） was used to analyze the effect of resveratrol on MCP-1 protein secretion during 3 T3-L1 differentiation. Results： Resveratrol significantly inhibited the proliferation of 3 T3-L1 preadipocytes and showed a dose-dependent effect. Compared with the control group,the resveratrol inhibited the synthesis of lipid droplets during the differentiation of 3 T3-L1 cells by inhibiting the expressions of PPARγ,C/EBPα and Notch1 mRNAs in adipocytes. The expression of Leptin mRNA in mature adipocytes induced by white reedosterol was decreased on the second day,and the expression of Leptin mRNA in the adipocytes of adipocytes induced by albinool was significantly lower than that of the control group（ P 〈0. 05） but increased on day 4. Conclusion： Resveratrol can inhibit the proliferation of 3 T3-L1 preadipocytes and preadipocyte differentiation. The mechanism may be through inhibition of PPARγ,C/EBPα and Notch1 expression. In the process of adipocyte differentiation of 3 T3-L1,resveratrol can promote the expression and secretion of MCP-1 during adipocyte differentiation and increase the expression of Leptin gene and regulate the effect of inflammation on obesity.