肺炎衣原体感染通过PI3K激活Rac1诱导血管平滑肌细胞迁移.docVIP

  肺炎衣原体感染通过 PI3K 激活 Rac1 诱导 血管平滑肌细胞迁移# 张俊霞，张腾腾，张利军，王蓓蓓，魏俊燕，王海伟，沈炳玲，张丽莙** 5 10 15 20 25 30 35 40 （天津医科大学基础医学院病理生理学教研室，天津 300070） 摘要：目的：研究 Rac1 活化在肺炎衣原体（C.pn）感染诱导血管平滑肌细胞（VSMC）迁 移中的作用及磷脂酰肌醇 3 激酶（PI3K）对其活化的影响。方法：GST-PBD 重组质粒转化 感受态细菌，诱导融合蛋白表达并纯化；GST-pull down 实验检测 C.pn 感染 VSMC 后 Rac1 活性的变化；PI3K 特异性抑制剂 LY294002 预处理 VSMC，GST-pull down 实验检测 Rac1 活性的变化；Rac1 特异性抑制剂 NSC23766 预处理 VSMC，Wound-healing 实验和 Transwell 实验观察 VSMC 迁移能力的变化。结果：重组质粒转化感受态细菌后，经诱导表达和纯化， 得到足量有效的 GST-PBD 融合蛋白；GST-pull down 实验结果显示，C.pn 感染 VSMC 后 Rac1 活性增强且显著高于正常对照组（P 0.05）；LY294002 预处理 VSMC 后，C.pn 感染 诱导的 Rac1 活性明显下降（P 0.05）；细胞迁移实验结果显示，NSC23766 预处理的 C.pn 感染组细胞迁移能力明显低于单纯感染组（P 0.05）。结论：C.pn 感染可能通过 PI3K 激 活 Rac1 诱导 VSMC 迁移。 关键词：肺炎衣原体；Rac1；PI3K；血管平滑肌细胞；细胞迁移 中图分类号：R363 Chlamydia pneumoniae infection induces vascular smooth muscle cell migration through stimulating Rac1 activity via PI3K activation ZHANG Junxia, ZHANG Tengteng, ZHANG Lijun, WANG Beibei, WEI Junyan, WANG Haiwei, SHEN Bingling, ZHANG Lijun (Department of Pathophysiology, Basic Medical College, Tianjin Medical University, TianJin 300070) Abstract: AIM: To detect the role of Rac1 activation in vascular smooth muscle cell (VSMC) migration induced by Chlamydia pneumoniae (C.pn) infection and determine the effect of Phosphoinositide 3-kinase (PI3K) on Rac1 activation during this process. METHODS: The recombinant plasmid encoding the GST-PBD fusion protein was transformed into the competent bacteria to induce the expression of the fusion protein. Then, the fusion protein was purified. GST-pull down assay was performed to evaluate the activity of Rac1 in C.pn-infected VSMCs pretreated with or without PI3K inhibitor LY294002. Wound-healing assay and Transwell assay were performed to observe the changes of C.pn infection-induced VSMCs migration with or without preincubation of Rac1 inhibitor NSC23766. RESULTS: After the purification of the GST-PBD fusion protein, the enough and biologically active fusion protein was obtained. After VSMCs were infected wit