中文摘要：

目的观察腺苷酸活化蛋白激酶（AMPK）a磷酸化在HepG2细胞脂变模型中的表达及意义。方法HepG2细胞予油酸和棕榈酸组成的0．3mmol／L游离脂肪酸（FFA）诱导24h后，建立脂肪变性HepG2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AMPKa和磷酸化AMPKa（pAMPKa）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），pAMPKa蛋白表达均显著降低（P〈0．01）。结论FFA诱导的脂肪变性HepG2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pAMPKtx蛋白的激活减少有关。

英文摘要：

Objective To establish a steatotic HepG2 cell model induced by free fatty acid（FFA） ,and observe the expression and significance of adenosine 5＂-monophosphate-activated protein kinase or（AIIPKot） phosphorylation in HepG2 cells of steatosis. Methods In- duced with DMEM medium of 0.3 mmol/L FFA（ composed of oleic acid and palmitic acid）for 24 hours, the steatotic HepG2 cell model Was successfully established（control group）. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red O staining, while the levels of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）in serum, and triglyceride （TG）ac- cumulation in HepG2 ceils were measured by automatic biochemical analyzer. The activities of superoxide dismutase （SOD） and malonyldial- dehyde （MDA）were tested by biological reagent kit, while the protein expression of AMPKa and pAMPKa were analyzed by Western blot. Re- suits Compared with those of control group,the levels of TG and MDA content were significantly higher （ P〈0.01 ）, and lots of red lipid droplets inside cytoplasm were visible, while the expression of pAMPKa protein and SOD levels were significantly reduced （P〈0.01）in mod- el group. Conclusions The FFA-induced steatosis in HepG2 cells may result in lipid metabolism disorder and oxidative stress, possibly by activating the pAMPKa protein mildly in HepG2 cells.