不同浓度同型半胱氨酸对内皮细胞TRPV1蛋白表达的影响

摘要

目的 研究不同浓度同型半胱氨酸对内皮细胞TRPV1蛋白表达水平和细胞增殖的影响.方法 将人脐静脉内皮细胞分别用不同浓度同型半胱氨酸(0、0.05、0.1、0.2、0.5、1、2、4、8和16mmol·L-1)及不同时间(6、12、24、36和48h)进行干预后用CCK-8检测细胞增殖及活性情况,选择合适的干预浓度进行下一步干预分组(0、0.1、0.5、1和2mmol·L-1),Western blot检测TRPV1蛋白表达水平.结果 1)同型半胱氨酸对人脐静脉内皮细胞的增殖能力有抑制作用,光密度(OD)值从(1.03±0.12)下降至(0.22±0.01),且呈剂量效应关系(F=11.73,P=0.000);2)同型半胱氨酸在0.5mmol·L-1开始、 干预24h后对人脐静脉内皮细胞增殖具有抑制作用(P<0.05);3)不同浓度同型半胱氨酸干预后TPRV1蛋白表达及细胞增殖能力低于对照组(P<0.05).结论 同型半胱氨酸对人脐静脉内皮细胞的增殖抑制具有剂量效应关系;随同型半胱氨酸浓度增加,TRPV1蛋白表达水平下降,细胞增殖能力亦降低.%Objective To explore the effects of homocysteine on the expression of TRPV1 and proliferation and in endothelial cell.Methods After human umbilical endothelial cells(HUVECs)intervened with different concentrations of homocysteine(Hcy)(0,0.05,0.1,0.2,0.5,1,2,4,8 and 16mmol·L-1)and at different times (6,12,24,36 and 48h),the proliferation and activity were detected using CCK-8 assay. Then HUVECs were intervened with proper concentrations of homocysteine(0,0.1,0.5,1 and 2mmol·L-1)based on the previous experiments and the expression of TRPV1 were assessed by Western blot.Results 1)The proliferation of HUVECs were significantly inhibited by homocysteine in a dose-dependent manner and the OD value was declined from(1.03±0.12)to(0.22±0.01)(F=11.73,P=0.000);2)Homocysteine intervention at 0.5mmol· L-1 for 24 hours had inhibiting effect on proliferation of human umbilical vein endothelial cells(P<0.05);3)The expression of TRPV1 and cell proliferation were significantly different from that of control group(P<0.05). Conclusion Homocysteine inhibits the proliferation of human umbilical vein endothelial cells in a dose-dependent mannnerand accompanied with downregulated of TRPV1.