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目的:观察激活的一磷酸腺苷活化蛋白激酶(AMPK)对软脂酸(PA)诱导的人脐静脉血管内皮细胞(HUVEC)损伤的保护作用,探讨其可能的作用机制。方法:按HUVEC培养液中成分不同,实验分为8组,分别为:空白对照组(常规培养)、PA培养组、5-氨基咪唑-4-甲酰胺核苷酸(AICAR)培养组、PA+AICAR培养组、二甲双胍(Met)培养组、PA+Met培养组、吡格列酮(PGZ)培养组和PA+PGZ培养组。HUVEC在各组培养液中分别培养24、48和72 h,MTT法测定不同时间点各组细胞的存活率,Western印迹法测定培养24 h时各组细胞的磷酸化AMPK蛋白表达水平。结果:与对照组相比,PA组细胞24、48和72 h的存活率明显降低,分别为58.95%、36.68%和15.09%(P<0.05);培养72 h,AICAR+PA组、Met+PA组和PGZ+PA组细胞的存活率均显著高于PA组(P<0.05);单独应用AICAR、Met和PGZ培养对细胞的存活率影响不大,与对照组比较无显著差异。与对照组和PA组相比,培养24 h时,AICAR、Met和PGZ刺激HUVEC的磷酸化AMPK表达增加(P<0.05)。结论:AICAR、Met和PGZ可能通过激活AMPK,显著减轻PA诱导的HUVEC的损伤。
Objective: To observe the protective effect of activated phosphorylated adenosine monophosphate kinase (AMPK) on the injury of human umbilical vein endothelial cells (HUVEC) induced by palmitic acid (PA) and to explore the possible mechanism. Methods: According to the different components of HUVEC culture medium, the experiment was divided into 8 groups: blank control group (conventional culture), PA culture group, AICAR culture group, PA + AICAR group, metformin group, PA + Met group, pioglitazone group and PA + PGZ group. HUVECs were cultured in each group for 24, 48 and 72 h respectively. The survival rates of cells in different groups at different time points were determined by MTT assay. The phosphorylated AMPK protein expression in each group was determined by Western blotting. Results: Compared with the control group, the survival rate of PA group was significantly lower at 24, 48 and 72 h (58.95%, 36.68% and 15.09%, respectively) The survival rates of PA and PGZ + PA group were significantly higher than that of PA group (P <0.05). AICAR, Met and PGZ alone had little effect on the survival rate of cells, but no significant difference compared with control group. Compared with the control group and the PA group, the phosphorylation of AMPK in HUVEC stimulated by AICAR, Met and PGZ increased (P <0.05) at 24 h. Conclusion: AICAR, Met and PGZ may significantly reduce PA-induced HUVEC injury by activating AMPK.