肉苁蓉苷F通过AMPK/ACC通路抑制C2C12细胞成脂分化

Cistanoside F inhibits the adipogenic differentiation of C2C12 cells through regulating AMPK/ACC pathway

目的探讨肉苁蓉苷F(Cis)抑制C2C12细胞成脂分化的效应及可能机制。方法C2C12细胞随机分为空白组、模型组、Cis低(1μmol/L)、中(10μmol/L)、高剂量(100μmol/L)组。CCK8检测细胞增殖率;油红O染色观察细胞内脂滴形成的动态变化;qRT-PCR检测生肌决定因子(Myod)、过氧化物酶体增殖激活受体(PPARγ)mRNA表达水平,Western Blot检测AMP依赖的蛋白激酶(AMPK)、乙酰辅酶A羧化酶(ACC)、结蛋白(Desmin)蛋白表达。结果各剂量Cis作用72h对C2C12细胞无明显毒性。与空白组相比,模型组细胞内脂滴在d8时显著增加,PPARγmRNA和ACC蛋白表达明显升高,Myod mRNA和AMPK、Desmin蛋白表达明显降低(P<0.05)。与模型组相比,Cis干预后,C2C12细胞内脂滴明显减少,PPARγmRNA表达下调,Myod mRNA表达上调,10μmol/L Cis作用8天可显著下调ACC蛋白表达,上调AMPK、Desmin蛋白表达(P<0.05)。结论Cis可能通过调控AMPK/ACC通路抑制C2C12细胞成脂分化。

Objective To explore the effect of Cistanoside F(Cis)on C2C12 cells adipogenic differentiation and the underlying mechanism.Methods C2C12 cells were randomly divided into vehicle group,model group,Cis low(1μmol/L),medium(10μmol/L)and high dose(100μmol/L)groups.The cell proliferation rate was detected by CCK8.Lipid drops at 2,4 and 8 days were observed by oil red O staining.The mRNA expression levels of Myogenic determination(Myod)and Peroxisome Prolif⁃erator-activated receptor gamma(PPARγ)were detected by qRT-PCR,the expressions of Adenosine 5′-monophosphate-ac⁃tivated protein kinase(AMPK),Acetyl CoA carboxylase(ACC)and Desmin were detected by Western Blot.Results Each dose of Cis had no significant effect on the proliferation rate of C2C12 cells.Compared with vehicle group,lipid droplets in C2C12 cells in model group were significantly increased,PPARγmRNA and ACC protein expressions were significantly increased,and Myod mRNA and AMPK,Desmin protein expressions were significantly down-regulated(P<0.05).Compared with model group,the content of lipid droplets in C2C12 cells in Cis groups was significantly decreased,PPARγmRNA was significantly down-regulated,and Myod mRNA expression was significantly up-regulated,10μmol/L Cis could significantly decrease the expression of ACC protein,and increase the expression of AMPK and Desmin protein(P<0.05).Conclusion Cis could inhibite the C2C12 cells adipogenic differentiation through regulating AMPK/ACC pathway.