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目的探讨软脂酸(PA)对人脐静脉内皮细胞凋亡的影响及其可能机制。方法 (1)将人脐静脉内皮细胞分为:①正常对照(NC)组;②PA组(浓度分别为0.2、0.4、0.8mmol/L),培养24h,采用MTT法检测细胞增殖情况,流式细胞术及Hoechst 33258核染色检测各组细胞凋亡情况。(2)人脐静脉内皮细胞分为:①NC组;②PA组(0.4mmol/L);③PA+雷帕霉素(PA+Rapamycin)组。采用West-ern blot法分析各组马铃薯球蛋白(Tuberin)、P-Tuberin、核糖体蛋白S6激酶(S6K)、P-S6K、Bcl-2、Bax,以及Caspase3蛋白表达水平。结果经MTT检测显示,与NC组比较,PA组细胞增殖(OD值)下降(P<0.05),且呈剂量依赖性。经Western blot分析显示,与NC组比较,PA组细胞凋亡率增加(P<0.05),且呈剂量依赖性。与NC组比较,PA组P-Tuberin、P-Tuberin/Tuberin、S6K、P-S6K/S6K、Bax、Caspase3表达水平增高,Bcl-2表达降低(P<0.05);PA+Rapamycin组S6K、P-S6K/S6K、Bax、Caspase3表达水平低于PA组,Bcl-2表达高于PA组(P<0.05)。结论 PA能够诱导人脐静脉内皮细胞凋亡,其机制可能是通过上调tuberin/mTOR活性,而降低Bcl-2表达、增加Bax及Caspase3表达,最终导致血管内皮细胞凋亡。
Objective To investigate the effect of palmitate (PA) on the apoptosis of human umbilical vein endothelial cells and its possible mechanism. Methods (1) The human umbilical vein endothelial cells were divided into: ① normal control (NC) group; ② PA group (0.2,0.4,0.8 mmol / L respectively), cultured for 24h, the cell proliferation was detected by MTT method, Cell apoptosis and Hoechst 33258 nuclear staining were used to detect the cell apoptosis. (2) Human umbilical vein endothelial cells were divided into: ①NC group; ②PA group (0.4mmol / L); ③PA + rapamycin group. Tuberin, P-Tuberin, S6K, P-S6K, Bcl-2, Bax and Caspase3 protein expression levels in each group were analyzed by West-ern blot. Results MTT assay showed that compared with NC group, the cell proliferation (OD) in PA group decreased (P <0.05) and in a dose-dependent manner. Western blot analysis showed that compared with NC group, the apoptosis rate of PA group increased (P <0.05), and in a dose-dependent manner. Compared with NC group, the expression of P-Tuberin, P-Tuberin / Tuberin, S6K, P-S6K / S6K, Bax and Caspase3 increased and the expression of Bcl-2 decreased (P < The expressions of S6K, S6K, Bax and Caspase3 were lower than those in PA group, while the expression of Bcl-2 was higher than that in PA group (P <0.05). Conclusions PA can induce the apoptosis of human umbilical vein endothelial cells by down-regulating the activity of tuberin / mTOR, decreasing the expression of Bcl-2, increasing the expression of Bax and Caspase3, and finally leading to the apoptosis of vascular endothelial cells.