板蓝根水提物对3T3-L1前脂肪细胞增殖和分化的影响

Effects of water extract of Radix Isatidis on proliferation and differentiation of 3T3-L1 preadipocytes

目的探讨板蓝根水提物(water extract of Radix Isatidis,WERI)对3T3-L1前脂肪细胞增殖分化的影响及其作用机制。方法体外培养3T3-L1前脂肪细胞,MTT法和流式细胞技术检测WERI对细胞增殖的影响;鸡尾酒诱导剂诱导3T3-L1前脂肪细胞,油红O染色和比色分析法观察WERI对前脂肪细胞分化及脂肪积累的影响;采用RT-PCR检测脂肪细胞的过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)及CCAAT/增强子结合蛋白α(CCAAT/enhancer-binding proteinα,C/EBPα)mRNA的表达;Western blot法检测PPARγ及C/EBPα蛋白表达。结果WERI在一定浓度范围内能有效抑制3T3-L1前脂肪细胞增殖,同时细胞周期呈现G_0/G_1向S期阻滞;与空白组相比,用不同浓度的WERI处理后,3T3-L1前脂肪细胞的分化受到明显抑制,且细胞内脂滴生成量明显减少;PCR及Western blot结果显示,WERI还可以抑制脂肪细胞PPARγ和C/EBPα基因及蛋白的表达。结论 WERI具有抑制3T3-L1前脂肪细胞增殖作用,并可通过下调PPARγ和C/EBPαmRNA及蛋白表达来抑制细胞成脂分化。

Aim To explore the effect of water extract of Radix Isatidis（ WERI） on proliferation and differentiation of 3T3-L1 preadipocytes. Methods 3T3-L1 preadipocytes were cultured to explore the effect of WERI on their proliferation measured by MTT and flow cytometry. Oil red O staining was applied to investigate the effect of different concentrations of WERI on the differentiation of 3T3-L1 preadipocytes. Intracellular fat accumulation was quantified using colorimetry.Semi-quantitative PCR was used to measure the expressions of PPARγ and C/EBPα mRNA. Western blot was applied to detect the expressions of PPARγ and C/EBPα. Results The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating with a certain range of concentration of WERI,and the cell cycle was also blocked from G0/G1 to S. After being treated with different concentrations of WERI,the differentiation of 3T3-L1 preadipocytes was significantly inhibited compared with control group,and the accumulation of lipid droplets within the cells were obviously decreased. It also down-regulated the expressions of PPARγ and C/EBPα both in mRNA and protein. Conclusion WERI can inhibit the proliferation and differ entiation of 3T3-L1 preadipocytes and reduce the accumulation of lipid drops,which may be associated with its effects in inhibiting the expressions of PPARγand C/EBPα both in mRNA and protein.