牛蒡子苷对L6骨骼肌细胞中腺苷酸活化蛋白激酶信号通路的影响

Effect ofarctiin on AMPK signal pathway in L6 myotubes

目的探讨牛蒡子苷在L6骨骼肌细胞中对葡萄糖消耗和腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)信号通路的调节作用。方法用含2%胎牛血清的1640培养基诱导L6骨骼肌细胞分化,直至细胞生长出肌管,分化完成后的L6骨骼肌细胞用含有不同浓度牛蒡子苷的培养基培养24 h后,采用葡萄糖氧化酶法检测细胞葡萄糖消耗,western blot法检测AMPK的亚单位AMPKα和磷酸化腺苷酸活化蛋白激酶(p-AMP-activated protein kinase,p-AMPKα)的蛋白表达水平;以real-time PCR法检测过氧化物酶体增生物激活受体γ共激活因子1α(peroxisome-proliferator-activated receptorγcoactivator-1α,PGC-1α)mRNA表达水平。结果牛蒡子苷浓度为1000 g/L时,可增加L6骨骼肌细胞的葡萄糖消耗,差异显著(P<0.01);与对照组相比,牛蒡子苷呈浓度依赖性的增强L6骨骼肌细胞中p-AMPKα蛋白表达水平;并以相同的趋势增强PGC-1αmRNA表达水平。结论牛蒡子苷增加L6骨骼肌细胞的葡萄糖消耗,并激活AMPK/PGC-1α信号通路,具有潜在的改善胰岛素抵抗的作用。

Objective To investigate the effect of arctiin on glucose consumption and AMPK pathway in L6 myotubes. Methods L6 myoblasts were differentiated to myotubes and treated with or without arctiin. After 24 h,glucose oxidase method was performed at the end of treatment and the AMPKα and p-AMPKα levels were detected by western blot（ mammalian AMPK is a trimer structure that composed of alpha,beta,gamma of three subunits,and the content detection of total AMPK and activated AMPK proteins usually refers to the alpha subunit.）; The PGC-1α mRNA level were measured with real-time PCR. Results The result of glucose oxidase assay suggested that arctiin increase. Compared with control group,arctiin enhanced the activity of p-AMPKα with a concentration-dependent manner,and arctiin promoted the transcription of PGC-1α with a concentration-dependent manner,too. Conclusion Arctiin increases glucose consumption and activates AMPK / PGC-1α signal pathway in L6 myotubes,which suggests that arctiin could potentially improve insulin resistance.