N-WASP在肺炎衣原体感染促进血管新生中的作用

Roles of N-WASP in angiogenesis promoted by Chlamydia pneumoniae infection

目的探讨N-WASP在肺炎衣原体感染诱导血管新生中的作用及其可能机制。方法肺炎衣原体增殖培养后感染人血管内皮细胞(VEC),免疫荧光染色确认感染成功。Western blot检测肺炎衣原体感染的VEC内N-WASP磷酸化水平;CCK-8检测N-WASP特异性抑制剂Wiskostain对VEC活力的影响;免疫荧光实验检测工作浓度的Wiskostain对肺炎衣原体感染率的影响;肺炎衣原体感染以5μmol/L Wiskostain预处理的VEC后,管腔形成实验观察各组VEC形成新生血管能力的变化。结果在荧光显微镜下,感染的VEC胞浆内可见典型的肺炎衣原体包涵体。肺炎衣原体感染VEC 10、24 h后N-WASP磷酸化水平均明显上调且高于正常对照组(P<0.05)。管腔形成实验结果显示,肺炎衣原体感染VEC 16 h后,其所形成的微管腔节点数明显多于正常对照组(P<0.05);经Wiskostain预处理后,肺炎衣原体感染促进微管腔节点形成的作用被显著削弱,几乎不能形成微管腔结构(P<0.05)。结论肺炎衣原体感染可能通过N-WASP促进人VEC形成新生血管。

Aim To investigate the roles of N-WASP in angiogenesis induced by Chlamydia pneumoniae （C.pneumoniae） infection and its possible mechanism. Methods After proliferation culture, C.pneumoniae infected human vascular endothelial cell （VEC）, and immunofluorescence staining confirmed successful infection. The phosphorylation level of N-WASP was detected by Western blot in VEC infected by C.pneumoniae. The effect of N-WASP specific inhibitor Wiskostain on VEC viability was detected by CCK-8. The effect of Wiskostain working concentrations on C.pneumoniae infection rate was detected by immunofluorescence assay. After C.pneumoniae infected VEC with 5 μmol/L Wiskostain pretreatment, capillary tube formation assay was performed to observe the changes of VEC angiogenesis ability. Results Under fluorescence microscope, typical C.pneumoniae inclusions were found in infected VEC cytoplasm. After VEC was infected with C.pneumoniae for 10 and 24 hours, the level of N-WASP phosphorylation was significantly higher than that in the control group. Capillary tube formation assay showed that after VEC was infected with C.pneumoniae for 16 hours, the number of capillary tube node was significantly higher than that in the control group （P〈0.05）. After the pretreatment of VECs with Wiskostain, the role of C.pneumoniae infection in promoting the formation of capillary tube node was significantly weakened, and it was almost impossible to form capillary tube structure （P〈0.05）. Conclusion C.pneumoniae infection may promote the formation of new blood vessels possibly through N-WASP.