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目的:观察华支睾吸虫溶血磷脂酶(CslysoPLA)重组蛋白体外对大鼠肝星状细胞和卵圆细胞的作用。方法:应用免疫荧光方法,观察CslysoPLA重组蛋白是否可与大鼠肝星状细胞和卵圆细胞膜结合;应用MTT方法,测定CslysoPLA重组蛋白对大鼠肝星状细胞和卵圆细胞的促增殖作用;应用流式细胞术(FCM)检测CslysoPLA重组蛋白对大鼠卵圆细胞细胞周期的影响。结果:CslysoPLA重组蛋白可与大鼠肝星状细胞和卵圆细胞膜结合,MTT方法测定结果表明低浓度重组蛋白对大鼠肝星状细胞和卵圆细胞有一定促增殖作用(P<0.05),20mg/L重组蛋白可促进大鼠卵圆细胞进入G2期,但高浓度重组蛋白可导致大鼠肝星状细胞和卵圆细胞死亡。结论:低浓度的CslysoPLA重组蛋白在体外可刺激大鼠肝星状细胞和卵圆细胞增殖,表明CslysoPLA可能是华支睾吸虫导致胆管上皮增生、腺瘤样病变和肝纤维化的效应分子之一。而高浓度的CslysoPLA重组蛋白可引起肝星状细胞和卵圆细胞死亡,表明CslysoPLA亦可能是华支睾吸虫引起的化学损伤的毒力因子之一。
Objective: To observe the effect of CslysoPLA recombinant protein on rat hepatic stellate cells and oval cells in vitro. Methods: Immunofluorescence method was used to observe whether CslysoPLA recombinant protein could bind to rat hepatic stellate cells and oval cell membrane. MTT assay was used to determine the effect of CslysoPLA recombinant protein on rat hepatic stellate cells and oval cells. The effect of CslysoPLA recombinant protein on the cell cycle of rat oval cells was analyzed by flow cytometry (FCM). Results: CslysoPLA recombinant protein could bind to rat hepatic stellate cells and oval cell membrane. The results of MTT assay showed that low concentration of recombinant protein could promote proliferation of rat hepatic stellate cells and oval cells (P <0.05) 20mg / L recombinant protein can promote rat oval cells into the G2 phase, but high concentrations of recombinant protein can cause rat hepatic stellate cells and oval cell death. CONCLUSIONS: Low concentration of CslysoPLA recombinant protein can stimulate the proliferation of rat hepatic stellate cells and oval cells in vitro, indicating that CslysoPLA may be one of the effector molecules of Clonorchis sinensis to cause biliary epithelial hyperplasia, adenomatous lesions and liver fibrosis . However, high concentrations of CslysoPLA recombinant protein can cause hepatic stellate cells and oval cell death, indicating that CslysoPLA may also be one of the virulence factors of chemical damage induced by Clonorchis sinensis.