C反应蛋白诱导内皮细胞炎症相关因子iNOS及ICAM-1表达的研究

Expression of inflammation related factors iNOS and ICAM-1 in endothelial cells induced by C-reactive protein

目的探讨不同浓度C反应蛋白(CRP)诱导内皮细胞炎症相关因子可诱导型一氧化氮合酶(i NOS)及细胞间黏附分子-1(ICAM-1)的表达及可能机制。方法实验分为对照组及1、5、10、20mg/L CRP处理组,分别对人脐静脉内皮细胞(HUVEC)进行相应浓度的CRP处理。采用RT-PCR法检测对照组及各CRP处理组HUVEC细胞的ICAM-1 mRNA表达变化;采用ELISA法检测1、5、20mg/L组CRP诱导的HUVEC中ICAM-1、i NOS的蛋白表达水平。结果 RT-PCR结果显示各组均有ICAM-1 mRNA表达,1、5、10mg/L CRP组与对照组比较,ICAM-1 mRNA表达差异无统计学意义,而20mg/L CRP组ICAM-1 mRNA的表达(1.125)为对照组(0.760)的1.48倍。ELISA检测结果显示1、5mg/L CRP组ICAM-1蛋白表达与对照组比较差异无统计学意义(P>0.05),而20mg/L CRP组与其他组比较,ICAM-1蛋白表达明显增加(P<0.01);1、5mg/L CRP组与对照组相比较,i NOS蛋白表达差异无统计学意义(P>0.05),而20mg/LCRP组与其他组比较,i NOS的蛋白表达明显增加(P<0.01)。结论 CRP在正常生理水平对内皮细胞的炎症相关因子表达影响不大,仅在较高浓度(>20mg/L)才可能对内皮细胞诱导炎症相关因子表达产生影响。

Objective To investigate the expression of inducible nitric oxide synthase（iNOS） and intercellular cell adhesion molecule-1（ICAM-1） in endothelial cells induced by C-reactive protein（CRP） and its corresponding mechanisms.Methods Human umbilical cord vein endothelial cells（HUVEC） were treated with different concentrations of CRP or with phosphate buffered solution as control,and RT-PCR was used for measurement of the expression of ICAM-1 mRNA induced by CRP in HUVECs.HUVEC were treated with CRP of 1mg/L,5mg/L,20mg/L,or with phosphate buffered solution,and expressions of ICAM-1 and iNOS protein in HUVECs were detected by cellular enzyme linked immunosorbent assay（ELISA）.Results In groups of 1mg/L,5mg/L and 10mg/L CRP,no different effects on expression of ICAM-1 mRNA in HUVECs was found when compared with control group,whereas the expression of ICAM-1 mRNA was elevated in the group of 20mg/L CRP by 1.48 folds compared with that in control group.Similarly,in groups of 1mg/L and 5mg/L CRP there was no significant difference in the expressions of ICAM-1 and iNOS in HUVECs compared with that in control group（P0.05）,whereas the expressions of ICAM-1 and iNOS protein were increased significantly in group of 20mg/L CRP compared with that in other groups（P0.01）.Conclusions Although CRP may induce the expression of inflammatory factors in endothelial cells,the present experioment showed that CRP had no significant effects on inflammatory factors in endothelial cells at normal physiological level,and it gave inducible effects at higher concentration（20mg/L） only.