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目的:研究激活TRPV1在高同型半胱氨酸促内皮细胞凋亡中的作用。方法:培养人脐静脉内皮细胞,用不同浓度同型半胱氨酸(0 mmol/L、0.05 mmol/L、0.1 mmol/L、0.2 mmol/L、0.5 mmol/L、1 mmol/L、2mmol/L、4mmol/L)及不同时间(6h、12h、24h、36h、48h)进行干预后CCK-8检测细胞增殖及活性情况,选择合适的干预浓度及干预时间;用免疫学方法检测TRPV1是否在人脐静脉内皮细胞上表达;Western blotting检测各组TRPV1蛋白的表达,TUNEL染色了解各组细胞凋亡情况。结果:(1)同型半胱氨酸对人脐静脉内皮细胞的增殖能力有明显的抑制作用,OD值从(1.03±0.12)渐下降至(0.42±0.05),且呈明显的剂量效应关系(P<0.01);同型半胱氨酸在0.5mmol/L开始、干预24h后对人脐静脉内皮细胞增殖有抑制作用(P<0.05);(2)细胞免疫荧光检测人脐静脉内皮细胞上有TRPV1表达,而对照组无表达;(3)不同浓度同型半胱氨酸干预细胞后TPRV1蛋白表达及细胞凋亡有差异(P<0.05);(4)辣椒素干预后TRPV1表达及细胞凋亡较单纯同型半胱氨酸干预组有统计学差异(P<0.05)。结论:高同型半胱氨酸可通过对TRPV1蛋白表达水平的下调,促进内皮细胞凋亡;激活TRPV1可抑制高同型半胱氨酸促内皮细胞凋亡。
Objective: To investigate the role of activating TRPV1 in the apoptosis of hyperhomocysteine-induced endothelial cells. Methods: Human umbilical vein endothelial cells were cultured and treated with different concentrations of homocysteine (0 mmol / L, 0.05 mmol / L, 0.1 mmol / L, 0.2 mmol / L, 0.5 mmol / L, L, 4mmol / L) and different time (6h, 12h, 24h, 36h, 48h) after intervention of CCK-8 detection of cell proliferation and activity, select the appropriate intervention concentration and intervention time; using immunological methods to detect whether TRPV1 Human umbilical vein endothelial cells were cultured. Western blotting was used to detect the expression of TRPV1 in each group. TUNEL staining was used to detect the apoptosis of each group. Results: (1) Homocysteine significantly inhibited the proliferation of human umbilical vein endothelial cells. The OD decreased gradually from (1.03 ± 0.12) to (0.42 ± 0.05), and showed a dose-dependent relationship P <0.01). Homocysteine inhibited the proliferation of human umbilical vein endothelial cells (P <0.05) at 0.5 mmol / L for 24 h; (2) (P <0.05); (4) The expression of TRPV1 and the apoptosis of TRPV1 after the intervention of capsaicin Compared with simple homocysteine intervention group, there was a significant difference (P <0.05). CONCLUSION: Homocysteine can promote the apoptosis of endothelial cells through the down-regulation of TRPV1 protein expression. Activation of TRPV1 can inhibit the apoptosis of hyperhomocysteine-induced endothelial cells.