血流剪切力对血管内皮细胞过敏毒素C3a受体表达影响的研究

The influence of shear stress on the expression of anaphylatoxin C3a receptor in vascular endothelial cells

目的:探讨血流剪切力对血管内皮细胞过敏毒素C3a受体(C3aR)、炎症因子及黏附分子表达的影响。方法:分离培养原代人脐静脉内皮细胞,利用振荡流(OS)刺激,分别于处理前、处理6小时、12小时、24小时收取细胞,用免疫印迹法检测C3a受体(C3aR)及血管细胞黏附蛋白1(VCAM 1)表达变化;用实时荧光定量检测C3aR、白介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)、细胞间黏附分子1(ICAM 1)及VCAM 1表达。结果:与处理前相比,内皮细胞在振荡流刺激后,C3a受体mRNA水平和蛋白表达均升高,6小时升高最显著分别为:[(1.997±0.3109)vs.(1.000±0.005773),P=0.0423;(2.968±0.2333)vs.(1.007±0.01202),P=0.0009];IL-1β、MCP-1、ICAM1及VCAM 1的mRNA水平均显著高于处理前,6小时达到最高值;TNF-α在刺激24小时后升高最显著(均P<0.05);振荡流刺激内皮细胞后VCAM 1表达上调,在6小时升高最显著(均P<0.05)。结论:血流剪切力可能通过上调血管内皮细胞过敏毒素C3a受体,调节内皮细胞炎症因子及黏附分子表达。

Objective： To determine the the influence of shear stress on the expression of anaphylatoxin C3 a receptor,inflammatory factors and adhesion proteins in vascular endothelial cells. Methods： Isolated and cultured the primary human umbilical vein endothelial cells,stimulated with the oscillatory shear stress and harvested the cells at 0 hour,6 hour,12 hour,and 24 hour after stimulation. Using western blot to observed the expression of C3 aR and adhesion protein VCAM 1. Using real time PCR to observe the expression of C3 aR,and inflammatory factors IL-1β,TNF-α,MCP-1,and adhesion protein VCAM 1 and ICAM 1. Results： In comparison with control,the expression of C3 aR on endothelial cells was significantly up-regulated after the stimulation of oscillatory shear stress,and the highest expression was at 6 hour（ mRNA level 6 hour vs. 0h was [（ 1. 997 ±0. 3109） vs.（ 1. 000 ± 0. 005773）,P = 0. 0423]; protein level 6 hour vs. 0 hour was [（ 2. 968 ± 0. 2333） vs.（ 1. 007 ± 0. 01202）,P = 0. 0009]. Real time PCR showed that the expression of IL-1β,MCP-1,ICAM 1and VCAM 1 were significantly up-regulated after stimulation,peaked at 6 hour. The expression of TNF-α was upregulated and peaked at 24 hour. Western blot showed that the protein level of VCAM 1 was significantly upregulated after stimulation,peaked at 6h. conclusion： Shear stress stimulate the expression of inflammatory fac-tors and adhesion proteins of endothelial cells,at least partially via up-regulation of anaphytoxin C3 aR.