尿毒症患者血清对内皮细胞小凹蛋白-1的影响及意义

Effects of Caveolin-1 in Human Umbilical Vein Endothelial Cell Induced by Serum of Uremia Patients

目的体外观察尿毒症患者血液透析前后血清和健康人血清对人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVECs)小凹蛋白-1(caveolin-1)表达的影响。方法取对数生长的HUVECs,分为健康组(DMEM+健康人血清,n=8)、透前组(DMEM+尿毒症患者透前血清,n=18)、透后组(DMEM+尿毒症患者透后血清,n=18)。采用MTT法检测细胞活力,免疫组化SABC法和Western blot检测各组细胞内caveolin-1蛋白的分布和含量。结果 MTT法筛选血清最佳干预时间和浓度分别为12小时、10%的血清干预浓度。与健康组比较,透前组细胞内caveolin-1的蛋白表达水平明显下调(p<0.05),透后组改变不明显(p>0.05);而透后组细胞内caveolin-1的蛋白表达水平较透前组细胞内caveolin-1的蛋白表达水平上调(p<0.05)。结论血液透析不能降低尿毒症患者动脉粥样硬化的形成的发生率,caveolin-1的变化可能是尿毒症血透患者动脉粥样硬化加速的原因之一。

Objective To detect the level of caveolin-1 protein expression in the human umbilical vein endothelial cells （HUVECs） induced by the serum of uremia patients before and after hemodialysis in vitro. Methods HUVECs in good condition were divided into 3 groups, healthy control group （ DMEM ＋ serum of healthy control, n = 8 ）, before dialysis group （ DMEM ＋ serum of ESRD patients before dialysis, n = 18） ,after dialysis group （ DMEM ＋ serum of ESRD patients after dialysis,n = 18）. Cell viability was examined by MTT. The levels of caveolin-1 protein expression were measured by western blot and immunohistochemistry. Results The best time was the twelfth hour and the best concentration of serum was 10% by MTT. Comparing with the healthy control group, the levels of caveolin-1 protein expression reduced significantly in before dialysis group （p 〈 0. 05 ）, but there is no significantly difference in after dialysis group （p 〉 0. 05 ）. Comparing with the before dialysis group, the levels of eaveoiln-1 protein expression improved notably in after dialysis group （p 〈 0.05 ）. Conclusion The expressions of eaveolin-1 in HUVECs are more in the after dialysis group than in the before dialysis group,and this change can become one of the important reasons why there is an increased ineidenee of atherosclerosis in the uremia patients on hemodialysis.