水飞蓟宾对C2C12细胞胰岛素抵抗的影响及机制研究

Study on Effects and Mechanisms of Silybin on Insulin Resistance of C2C12 Cells

目的观察水飞蓟宾对棕榈酸诱导C2C12肌管细胞胰岛素抵抗模型葡萄糖摄取能力的影响,并从STAT3/SOCS3信号通路探讨其作用机制。方法用含2%马血清的高糖培养基(DMEM)培养小鼠C2C12细胞4天后,改用0.5 mmol·L^(-1)棕榈酸造模液继续诱导培养16 h建立C2C12细胞胰岛素抵抗模型;用CCK-8法测定水飞蓟宾对C2C12细胞增殖能力的影响;用葡萄糖氧化酶法检测水飞蓟宾对C2C12细胞葡萄糖摄取能力的影响;用酶联免疫吸附法(ELISA)检测C2C12细胞培养液中炎性因子TNF-α、IL-β和IL-10的水平;用免疫印迹法(Western blot)检测水飞蓟宾对P-STAT3、SOCS3蛋白表达水平的影响。结果水飞蓟宾浓度在≥30μmol·L^(-1)时显著抑制C2C12细胞增殖;水飞蓟宾可增加C2C12细胞对葡萄糖的摄取,降低致炎因子TNF-α,IL-β,提高抑炎因子IL-10水平,同时抑制P-STAT3、SOCS3蛋白表达。结论水飞蓟宾可改善C2C12细胞胰岛素抵抗,其作用机制可能通过抑制TNF-α、IL-β炎症因子分泌,增加抑炎因子IL-10分泌,调控STAT3/SOCS3信号实现。

Objective To observe effects of silybin on glucose uptake in insulin resistance models of C2 C12 myotube cells induced by palmitic acid, and to explore its mechanism from a perspective of STAT3/SOCS3 signaling pathways.Methods Mouse C2 C12 cells were cultured in a high glucose medium(DMEM) with 2% horse serum for 4 days, then0.5 mmol·L^(-1) palmitic acid modeling solution was used to continue inducing and culturing for 16 h to establish insulin resistance models of the C2 C12 cells. The CCK-8 method was used to determine effects of silybin on proliferation of the C2 C12 cells. Effects of silybin on glucose uptake of the C2 C12 cells were measured by the glucose oxidase method.Levels of inflammatory factors TNF-α, IL-β and IL-10 in the C2 C12 cell culture medium were detected by enzyme linked immunosorbent assay(ELISA). The Western blot method was adopted to detect effects of silybin on expression levels of P-STAT3 and SOCS3 proteins.Results Silybin could significantly inhibit the proliferation of C2 C12 cells when the concentration was ≥ 30 μmol·L^(-1). Silybin could increase the glucose uptake of C2 C12 cells, decrease the levels of inflammatory factors TNF-α and IL-β, increase the level of the inflammatory factor IL-10, and inhibit the expressions of P-STAT3 and SOCS3 proteins.Conclusion Silybin can improve the insulin resistance of C2 C12 cells, and its mechanism may be related to inhibiting secretion of the inflammatory factors TNF-α and IL-β, increasing secretion of the factor IL-10 that inhibits inflammation, and regulating the STAT3/SOCS3 signaling pathways.