中国病理生理杂志2011，Vol.27Issue(9)：1682-1686,5.DOI:10.3969/j.issn.1000-4718.2011.09.004
赖型钩端螺旋体溶血素HlyX对血管内皮细胞通透性的影响
Effect of hemolysin protein HlyX of Leptospira interrogans serovar Lai on permeability of vascular endothelial cells
摘要
Abstract
AIM: To clone and express the hemolysin gene hlyX of Leptospira interrogans serovar Lai and to investigate the effect of the expression product on the permeability of human umbilical vein endothelial cells ( HUVECs ).METHODS: The recombinant plasmid pET - hlyX was constructed by inserting the hlyX gene into prokaryotic expression vector pET32a( + ), and transformed into E.coli BL21( DH3 ) to express the fusion protein Trx - HlyX with a His - tag.The fusion protein was purified using HisTrap affinity columns.The permeability of the monolayer HUVECs was measured by enzyme - linked immunosorbent assay for biotin - labeled albumin.Flow cytometry and Hoechst 33258 staining were applied to measure the apoptotic rate of HUVECs after incubation with Trx - HlyX.RESULTS: The recombinant plasmid pET - HlyX was successfully constructed and the fusion protein Trx - HlyX was highly expressed.Compared with the control cells, the purified recombinant protein Trx - HlyX significantly increased the permeability of transfected cells and promoted apoptosis of HUVECs ( P ＜0.05 ).CONCLUSION: The recombinant plasmid pET - hlyX highly expresses the fusion protein Trx - HlyX.Purified protein Trx - HlyX influences the permeability and has cytotoxicity on HUVECs.关键词
钩端螺旋体/HlyX/通透性/细胞凋亡/人脐静脉内皮细胞Key words
Leptospira interrogans/ HlyX/ Permeability/ Apoptosis/ Human umbilical vein endothelial cells分类
医药卫生引用本文复制引用
孙湛,吴秉婷,李道坤,鲍朗..赖型钩端螺旋体溶血素HlyX对血管内皮细胞通透性的影响[J].中国病理生理杂志,2011,27(9):1682-1686,5.基金项目
国家自然科学基金资助项目(No.30771908) （No.30771908）
