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目的比较饱和脂肪酸棕榈酸(palmitic acid,PA)和单不饱和脂肪酸油酸(oleic acid,OA)对HepG2细胞凋亡和自噬的影响。方法 PA和OA处理HepG2细胞,CCK-8检测脂肪酸的细胞毒性,caspase-3活性测定检测脂肪酸对细胞凋亡的影响,油红O染色检测细胞内脂质沉积,酶法检测细胞内甘油三酯的含量,Western blot检测自噬相关蛋白LC3-Ⅱ和凋亡相关蛋白caspase-3的表达,实时荧光定量PCR检测相关基因表达。结果 PA和OA都均同时诱导HepG2细胞凋亡与自噬,但PA的影响明显强于OA;PA诱导HepG2细胞自噬相关基因LC3B和VPS38表达、促进脂肪酸β-氧化的PPARα基因mRNA水平升高,但对细胞内脂肪沉积没有明显作用;而OA则导致HepG2细胞内甘油三酯的水平明显增加,但对上述基因表达没有明显影响。结论饱和脂肪酸PA和不饱和脂肪酸OA均能引起HepG2细胞凋亡和自噬,但PA的作用强于OA。
Objective To compare the effects of saturated fatty acid palmitic acid (PA) and monounsaturated fatty acid oleic acid (OA) on apoptosis and autophagy in HepG2 cells. Methods HepG2 cells were treated with PA and OA. CCK-8 was used to detect the cytotoxicity of fatty acids. The effect of fatty acids on apoptosis was assayed by caspase-3 activity assay. Lipid deposition was detected by oil red O staining and triglyceride Western blot was used to detect the expression of autophagy-related protein LC3-Ⅱ and apoptosis-related protein caspase-3, and the expression of related genes was detected by real-time fluorescence quantitative PCR. Results Both PA and OA induced apoptosis and autophagy in HepG2 cells at the same time, but the effect of PA was stronger than that of OA. PA induced the expression of autophagy-related genes LC3B and VPS38 in HepG2 cells and increased the mRNA level of PPARα in fatty acid β-oxidation , But had no significant effect on intracellular fat deposition. However, OA caused a significant increase in triglyceride levels in HepG2 cells, but had no significant effect on the expression of these genes. Conclusion Both saturated fatty acid PA and unsaturated fatty acid OA can induce HepG2 cell apoptosis and autophagy, but the effect of PA is stronger than that of OA.