烟曲霉感染后人脐静脉内皮细胞表达组织因子的变化

Expression of the tissue factor in human umbilical vein endothelial cells infected with aspergillus fumigatus hypha

目的通过建立烟曲霉菌丝感染人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)的体外模型,观察烟曲霉菌丝感染后HUVEC表达组织因子(tissue factor,TF)的变化,以及用不同干预因素处理对HUVEC表达TF的影响,以探讨侵袭性肺曲霉病(invansive pulmonary aspergillosis,IPA)血管侵袭的机制。方法实验分为6组,即对照组、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)组、烟曲霉菌丝组、细胞松弛素D组、N-cadherin抗体组和地塞米松组,分别于0、2、6、12、18h获取细胞冻融液,采用ELISA法检测TF浓度。结果实验组2、6、12、18h TF表达量与0h比较均升高(P(0.05),而对照组则无明显变化;实验组TF表达量在2、6、12、18h高于对照组(P(0.05);TNF-α组的TF表达量在2、6、12h高于烟曲霉菌丝组(P(0.01);N-cadherin抗体组的TF表达量在2、6、12h低于烟曲霉菌丝组(P(0.05);细胞松弛素D组或地塞米松组的TF表达量在各时间点与烟曲霉菌丝组比较差异均无统计学意义。结论烟曲霉菌丝感染后,HUVEC过度表达TF,N-cadherin单克隆抗体可减少TF的表达,而细胞松弛素D或地塞米松对TF的表达无明显影响。

Objective The aim of this study was to explore the mechanism of angioinvasion in invasive pulmonary aspergillosis by observing the expression levels of the tissue factor（TF） in human umbilical vein endothelial cells（HUVECs）following aspergillus fumigatus hypha（AFH） infection or after intervention with various substances using an in vitro model of HUVEC infection with aspergillus fumigatus hypha.Methods Six groups were involved in our experiment： negative control,TNF-α,AFH,cytochalasin D,N-cadherin antibody and dexamethasone.The liquid of cell freezing and thawing was obtained at 0,2,6,12 and 18 hours of cultivation with the specific substances,and the expression levels of TF were detected in different groups by ELISA.Results Compared with the TF level at 0 hour,those at 2,6,12 and 18 hours were significantly increased（P0.05） in all but the negative control group.The TF level was markedly lower in the negative control than in the other experimental groups at 2,6,12 and 18 hours（P0.05）.And at 2,6 and 12 hours,the AFH group showed an obviously lower level of TF than the TNF-α（P0.01）,but a higher level than the N-cadherin antibody group（P0.05）.No statistically significant differences were observed between the AFH group and the cytochalasin D or dexamethasone group at any time point.Conclusion TF is overexpressed in AFH-infected HUVECs.The N-cadherin monoclonal antibody can reduce the expression of TF,but neither cytochalasin D nor dexamethasone has any significant effect on it.