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目的:研究共轭亚油酸(conjugated linoleic acid,CLA)对HepG2细胞脂肪沉积的影响,以探讨其对肝脏脂肪沉积的可能机制。方法:体外培养HepG2细胞,不同浓度CLA(0、10、50、100μmol.l-1)作用24小时后,100nmol.l-1胰岛素作用1小时。以比色法测定培养液中游离脂肪酸的含量,油红O染色观察细胞脂肪沉积,Western-blot检测Akt和P-Akt蛋白表达。结果:随着CLA浓度增加,培养液中的游离脂肪酸含量显著降低,细胞中红染脂滴增多,Western-blot结果发现,Akt蛋白的磷酸化水平增高。结论:CLA可以增加细胞内脂肪合成和细胞中Akt蛋白的磷酸化水平。
OBJECTIVE: To study the effects of conjugated linoleic acid (CLA) on the fat deposition in HepG2 cells in order to investigate its possible mechanism of hepatic fat deposition. Methods: HepG2 cells were cultured in vitro. After treated with different concentrations of CLA (0, 10, 50 and 100 μmol·L-1) for 24 hours, 100 nmol.l-1 insulin was administered for 1 hour. The content of free fatty acids in the culture medium was determined by colorimetric method. The fat deposition was observed by oil red O staining. The protein expression of Akt and P-Akt was detected by Western-blot. Results: With the increase of CLA concentration, the content of free fatty acids in the culture medium decreased significantly, and the amount of red dye in the cells increased. Western blot results showed that the phosphorylation level of Akt protein increased. Conclusion: CLA can increase intracellular fat synthesis and phosphorylation of Akt in cells.