Ma Chang-Ling, Hu Xu-Chu, Hu Feng-Yu, Zhou Hong-Juan, Xue Ling, Yu Xin-Bing
Department of Microbiology and Parasitology, Guangzhou Medical College, Guangzhou 510182, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jul;24(7):692-5.
To clarify the effects of the recombinant protein of Lysophospholipase from Clonorchis sinensis (CsLysoPLA) on the hepatic stellate cells (HSC) and oval cells of rat.
Binding of the recombinant CslysoPLA protein to the membrane of HSC and oval cells was identified by immunofluorescent staining. The HSC and oval cells were cultured and treated with the recombinant protein at different doses, and proliferation was quantified by MTT method. Cell cycle analysis was performed by flow cytometry.
The recombinant CslysoPLA protein could bind to the membrane of HSC and oval cells. Compared to control, 2 mg/L and 20 mg/L the recombinant protein could promote HSC and oval cells growth (P<0.05), whereas 200 mg/L the recombinant protein could induce the cells necrosis, which associated with overt plasma membrane disruption. Oval cell number in G(2) phase of the recombinant protein 20 mg/L treated group was higher than that of control group.
In vitro, the recombinant protein could induce HSC and oval cells proliferation at low concentrations (2 mg/L and 20 mg/L), whereas it also could induce the cells necrosis at high concentration (200 mg/L). These results suggested that CslysoPLA might play a role in the pathogenicity of C. sinensis.
阐明华支睾吸虫溶血磷脂酶重组蛋白(CsLysoPLA)对大鼠肝星状细胞(HSC)和卵圆细胞的影响。
采用免疫荧光染色鉴定重组CslysoPLA蛋白与HSC和卵圆细胞膜的结合情况。培养HSC和卵圆细胞,并用不同剂量的重组蛋白处理,通过MTT法对细胞增殖进行定量分析。采用流式细胞术进行细胞周期分析。
重组CslysoPLA蛋白能与HSC和卵圆细胞膜结合。与对照组相比,2 mg/L和20 mg/L的重组蛋白可促进HSC和卵圆细胞生长(P<0.05),而200 mg/L的重组蛋白可诱导细胞坏死,这与明显的质膜破坏有关。重组蛋白20 mg/L处理组处于G(2)期的卵圆细胞数量高于对照组。
体外实验中,低浓度(2 mg/L和20 mg/L)的重组蛋白可诱导HSC和卵圆细胞增殖,而高浓度(200 mg/L)时则可诱导细胞坏死。这些结果提示CslysoPLA可能在华支睾吸虫的致病性中发挥作用。