N-钙黏蛋白抗体对毛霉菌丝感染人肺微血管内皮细胞组织因子和血管性血友病因子表达水平的影响

Effect of N-cadherin Antibody on TF and vWF Expression by HPMVEC Infected with Mucor Irregularis

目的:观察毛霉菌丝对人肺微血管内皮细胞(HPMVEC)分泌凝血因子的影响及N-钙黏蛋白抗体的干预作用。方法:取增殖期HPMVEC,分为毛霉菌丝感染模型组(毛霉菌丝组,用1×104 CFU毛霉菌丝与HPMVEC在37℃、5%CO_2饱和湿度下共培养12h)、人N-钙黏蛋白单克隆抗体干预组[N-钙黏蛋白抗体组,在HPMVEC与毛霉菌丝共培养前,先用该抗体(1mg/ml)与HPMVEC室温作用1h,之后按毛霉菌丝组操作]和空白对照组(用无菌去离子水代替毛霉菌丝悬液,且不行N-钙黏蛋白抗体作用,按毛霉菌丝组方法平行实验)。分别于0h、1h、2h、6h、12h收集各组培养上清液,采用ELISA检测其组织因子(TF)和血管性血友病因子(vWF)水平。结果:组内比较,毛霉菌丝组TF和vWF水平随培养时间增加而升高(均P<0.05或P<0.01);组间比较,毛霉菌丝组各时相TF和vWF水平均较空白对照组升高(P<0.05或P<0.01),而N-钙黏蛋白抗体组各时相TF和vWF水平均较毛霉菌丝组显著降低(P<0.05或P<0.01)。结论:毛霉菌丝感染HPMVEC所致TF、vWF水平升高可被N-钙黏蛋白单克隆抗体有效下调,或可用于防治由毛霉菌丝感染引起的血管内皮细胞损伤和血栓形成。

Objective：To investigate the effect of mucor hypha on human pulmonary microvascular endothelial cell（HPMVEC）and the intervention of N-cadherin antibody.Method：The proliferative phase of HPMVEC,divided into mucor infection model group（mucor hypha group,1×104 CFU mucor hypha and HPMVEC co-culture at37℃,5% CO2 and saturated humidity for 12h）,N-cadherin monoclonal antibody intervention group（N-cadherin antibody group,HPMVEC and 1mg/ml N-cadherin antibody co-culture at room temperature for 1h,before the infection of mucor hypha,then followed by mucor hypha group operation）and blank control group（sterile deionized water instead of mucor hypha,and the absence of the N-cadherin antibody,then followed the mucor hypha group method of parallel experiment.The supernatant of each group was collected at 0h,1h,2h,6hand 12 h,and the levels of TF and vWF were detected by ELISA.Results：Comparison with groups,the levels of TF and vWF secreted in the supernatants of the mucor hypha group were significantly increased as time-dependent（P〈0.05 or P〈0.01）.Comparison between groups,the levels of TF and vWF secreted in the supernatants of the mucor hypha group were significantly higher than the blank control group at each time point（P〈0.05 or P〈0.01）,and after incubated with N-cadherin monoclonal antibody,the levels of TF and vWF released in the supernatants were significantly lower than the untreated mucor hypha group at each time point（P〈0.05 or P〈0.01）.Conclusion：The increased level of TF and vWF induced by mucor hypha infected with HPMVEC,which can be effectively down-regulated by N-cadherin monoclonal antibody,it suggested the N-cadherin monoclonal antibody can be used to prevent the injury and thrombosis of vascular endothelial cells caused by infection of mucor hypha.