辣木异硫氰酸酯改善C2C12肌管细胞胰岛素抵抗

Isothiocyanate Extracted from Moringa oleifera Improves Insulin Resistance of C2C12 Myotube Cells

目的:建立C2C12肌管细胞胰岛素抵抗(C2C12-IR)模型,探讨辣木异硫氰酸酯(MIC-1)对C2C12肌管细胞胰岛素抵抗的影响。方法:利用棕榈酸(PA)诱导C2C12肌管细胞,建立稳定的C2C12-IR模型,并用不同浓度MIC-1(0,2,4,8,16μmol/L)处理细胞16 h。采用MTT法检测C2C12-IR细胞存活率,并观察MIC-1对C2C12-IR细胞葡萄糖代谢的影响;检测细胞氧化损伤情况,包括一氧化氮(NO)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH);采用Western blot检测磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路中相关蛋白表达变化情况。结果:确定了C2C12-IR模型最佳建模条件,即用0.5 mmol/L PA处理C2C12肌管细胞16 h;与C2C12-IR组相比,MIC-1呈剂量依赖性方式,显著增加了C2C12-IR细胞葡萄糖消耗量和糖原含量;MIC-1显著降低了C2C12-IR细胞中MDA和NO水平,显著增加了GSH水平;此外,MIC-1显著增加了丙酮酸脱氢酶激酶同工酶1(PDK1)、磷酸化AKT和葡萄糖转运蛋白4(GLUT4)的表达水平。结论:MIC-1通过抑制氧化应激改善胰岛素抵抗。

Objective:A C2C12 myotube cell insulin resistance(C2C12-IR)model was established to explore the effect of isothiocyanate extracted from Moringa oleifera(MIC-1)on C2C12 myotube cell insulin resistance.Methods:Palmitic acid(PA)was used to induce C2C12 myotube cells to establish a stable C2C12-IR model,and the cells were treated with different concentrations of MIC-1(0,2,4,8,16μmol/L)for 16 h.The cell viability of C2C12-IR cells was detected by MTT assay,and the effect of MIC-1 on glucose metabolism of C2C12-IR cells was observed.Cellular oxidative damage[nitric oxide(NO),malondialdehyde(MDA)and glutathione peroxidase(GSH)]were tested.The expression levels of related proteins in the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway were detected by using Western blot assay.Results:The optimal modeling conditions for the C2C12-IR model were determined,that is,C2C12 myotube cells were treated with 0.5 mmol/L PA for 16 h.Compared with the C2C12-IR group,MIC-1 significantly increased the glucose consumption and glycogen content of C2C12-IR cells in a dose-dependent manner.MIC-1 significantly decreased the level of MDA and NO,and significantly increased the level of GSH in C2C12-IR cells.In addition,MIC-1 significantly increased the expression levels of phosphoinositide-dependent protein kinase-1(PDK1),p-AKT,and glucose transporter 4(GLUT4).Conclusion:MIC-1 improves insulin resistance by inhibiting oxidative stress.