目的 研究LA0202为孔形成溶血素.方法 体外克隆表达LA0202蛋白.纯化的LA0202蛋白作用绵羊血细胞观察LA0202的溶血活性并用渗透压保护剂PEG6000添加到作用体系研究渗透压保护剂对LA0202溶血活性的影响.透射电镜及扫描电镜观察LA0202作用于体外培养的肝细胞后对细胞的毒性.结果 渗透压保护剂PEG6000能够抑制LA0202的溶血活性.LA0202作用于肝细胞后引起肝细胞的毒性损伤.结论 LA0202为一孔形成溶血素.%Hemolysins of Leptospira interragans have been shown to be the virulence factor in the pathogenesis of leptospirosis and 10 potential hemolysin genes were charecterized by genomic annotation of L.interrogans serovar.Lai strain 56601. In the present study, the LA0202 gene supposed to encode one of the new potential hemolysin was cloned and the protein encoded was purified. The purified protein was shown to have highly hemolytic activity as demonstrated on the sheep blood agar plate. It was also confirmed that the LA0202 protein-mediated hemolysis on sheep erythrocytes was osmotically protected by PEG6000. Meanwhile, this protein could induce pore formation on sheep erythrocytes and cause damages on the membrane of human L-02 liver cells. In addition, it could induce apoptosis of human L-02 liver cells after treatment of cells with this protein for 24 hours. It is evident that LA0202 protein acting as a pore-formong hemolysin can induce cytotoxic damage on mammalian cells.
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