激活TRPV1在高同型半胱氨酸促内皮细胞凋亡中的作用

发布时间：2018-12-29 18:57

【摘要】：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(P0.01);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(P0.05);(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(P0.05);(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(P0.05)。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。
[Abstract]:Aim: to study the effect of activating TRPV1 on the apoptosis of endothelial cells induced by homocysteine. Methods: human umbilical vein endothelial cells were cultured with different concentrations of homocysteine (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) and different time (6 h ~ 12 h ~ 24 h ~ 36 h ~ 48 h) were used to detect the proliferation and activity of CCK-8, and the appropriate intervention concentration and intervention time were selected. The expression of TRPV1 on human umbilical vein endothelial cells (HUVECs) was detected by immunological method. The expression of TRPV1 protein in each group was detected by TUNEL staining and apoptosis was detected by TUNEL staining. Results: (1) homocysteine significantly inhibited the proliferation of human umbilical vein endothelial cells, and the OD value decreased from (1.03 卤0.12) to (0.42 卤0. 05) in a dose-dependent manner (P0.01). Homocysteine inhibited the proliferation of human umbilical vein endothelial cells (P0.05); (2) at the beginning of 0.5mmol/L and 24 h after intervention (P0.05); (2). The expression of TRPV1 in human umbilical vein endothelial cells was detected by immunofluorescence, but no expression was found in the control group. (3) the expression of TPRV1 protein and cell apoptosis were different in different concentrations of homocysteine (P0.05). (4) the expression of TRPV1 and apoptosis after capsaicin intervention were significantly higher than those in homocysteine group (P0.05). Conclusion: high homocysteine can promote the apoptosis of endothelial cells by down-regulating the expression of TRPV1 protein, and activating TRPV1 can inhibit the apoptosis of endothelial cells induced by high homocysteine.
【作者单位】：宁夏医科大学总医院心脏中心;宁夏医科大学心脑血管病医院;宁夏医科大学;宁夏医科大学总医院心研所;
【基金】：宁夏医科大学优势学科群建设科研项目(No:XY201501) 宁夏自然科学基金(No:NZ14113)
【分类号】：R54

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