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目的观察腺苷酸活化蛋白激酶(AMPK)α磷酸化在Hep G2细胞脂变模型中的表达及意义。方法 Hep G2细胞予油酸和棕榈酸组成的0.3 mmol/L游离脂肪酸(FFA)诱导24 h后,建立脂肪变性Hep G2细胞模型,并设置对照组比较。诱导成功后,采用油红O染色观察细胞内脂滴蓄积状态;采用全自动生化仪检测细胞上清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)含量和细胞内甘油三酯(TG)含量;采用生物试剂盒检测细胞内超氧化物歧化酶(SOD)和丙二醛(MDA)含量的变化,运用Western印迹法检测各组细胞AMPKα和磷酸化AMPKα(p AMPKα)蛋白的表达。结果与对照组比较,模型组细胞内橘红色脂滴大量形成,且细胞内TG和MDA含量明显升高(P<0.01),SOD含量水平明显下降(P<0.01),p AMPKα蛋白表达均显著降低(P<0.01)。结论 FFA诱导的脂肪变性Hep G2细胞模型可以出现脂质代谢紊乱和氧化应激状态,其机制可能与细胞内p AMPKα蛋白的激活减少有关。
Objective To observe the expression and significance of AMPK α phosphorylation in Hep G2 cell steatosis model. Methods Hep G2 cells were induced by 0.3 mmol / L free fatty acid (FFA) consisting of oleic acid and palmitic acid for 24 h, then the model of steatosis Hep G2 was established and compared with the control group. After successful induction, lipid red O staining was used to observe the intracellular lipid accumulation. The contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and intracellular The content of triglyceride (TG) was measured. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in cells were detected by using biological kit. The expression of AMPKα and p-AMPKα ) Protein expression. Results Compared with the control group, the lipid droplets in the model group were formed in large numbers and the contents of TG and MDA in the model group were significantly increased (P <0.01) and the content of SOD was significantly decreased (P <0.01), and the protein expression of p AMPKα in the model group was significantly Decreased (P <0.01). Conclusion FFA-induced steatosis Hep G2 cell model may appear lipid metabolism disorders and oxidative stress, the mechanism may be related to the reduction of intracellular p AMPKα protein activation.