鞣花酸对3T3-L1前体脂肪细胞增殖与分化的影响

The effect and mechanism of ellagic acid on proliferation and differentiation in 3T3-L1 preadipocytes

目的探讨鞣花酸对3T3-L1前体脂肪细胞增殖、分化的影响及其作用机制。方法培养3T3-L1前体脂肪细胞,用不同浓度的鞣花酸(10、15、20μmol/L)进行干预,并设对照组(无鞣花酸)。WST-1法观察不同浓度的鞣花酸对3T3-L1前体脂肪细胞增殖的影响;Western blot法检测鞣花酸对CCAAT/增强子结合蛋白β(C/EBPβ)总蛋白(LAP亚型)表达及C/EBPβThr188位点磷酸化的影响。结果鞣花酸可浓度依赖性抑制3T3-L1前体脂肪细胞增殖;与对照组比较,鞣花酸抑制C/EBPβThr188位点磷酸化(P<0.05),不同时间段抑制率分别为25.9%(2h)、37.1%(4h)、27.8%(8h)和25.2%(16h);对C/EBPβ总蛋白(LAP亚型)表达无影响。结论鞣花酸能抑制3T3-L1前体脂肪细胞增殖和分化,其机制可能是通过抑制C/EBPβThr188位点磷酸化,抑制脂肪形成过程中有丝分裂扩增而实现。

Objective To investigate the potential effects and the underlying mechanism（s） of ellagic acid （EA） on proliferation and differentiation of 3T3-L1 preadipocytes. Methods The 3T3-L1 preadipocytes were cultured and exposed to EA at the dose of 10, 15, 20μmol/L respectively. Those cells without EA served as placebo controls. Cell proliferation was determined by WST-1 staining. The protein expression of the total CCAAT-enhancer binding proteins 13 （C/EBP13, LAP） and phospho-C/EBPβ （Thr188） was detected by Western Blot. Results EA inhibited the proliferation of 3T3-L1 preadipocytes in a concentration-dependent manner. Additionally, EA prevented C/EBP13 Thr188 phosphorylation during adipogenesis. And the inhibition rate was 25.9%（2 h）, 37.1% （4 h）, 27.8%（8 h） and 25.2%（16 h）, respectively at different time points. EA showed no significant effect on the total C/EBPβ protein expression. Conclusion EA can inhibit preadipocyte differentiation and proliferation. And the mechanism may be related to its preventing C/EBPβ Thr188 phosphorylation and inhibiting mitotic amplification during adipogenesis.