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目的 探讨东方田鼠E77.43基因片段治疗日本血吸虫感染的生物学功能。方法 构建pRevTRE-E77.43重组逆转录病毒载体,转染PA317细胞,用Hygromycin筛选后获得稳定表达的细胞株,并进行重组逆转录病毒的包装、浓缩与纯化。通过静脉或腹腔途径将制备的重组病毒注射到日本血吸虫感染模型小鼠体内,探讨目的基因在小鼠体内的表达以及体内治疗血吸虫感染的作用。结果 病毒注射动物机体7 d后目的基因就有表达,且一直能持续表达45 d以上。pRevTRE-E77.43诱导小鼠产生了31.0%的减虫率,高于pRevTRE的1.2%(t=3.524,P<0.01);pRevTRE-E77.43诱导小鼠产生了35.0%的减卵率,高于pRevTRE的0.9%(t=9.485,P<0.01)。结论 pRevTRE-E77.43能对血吸虫感染模型小鼠产生一定程度的治疗效果,说明东方田鼠E77.43基因参与了东方田鼠抗日本血吸虫感染的生物学作用过程。
Objective To investigate the biological function of Euplexurium viperns E77.43 gene in the treatment of Schistosoma japonicum infection. Methods The recombinant retroviral vector pRevTRE-E77.43 was constructed and transfected into PA317 cells. The stable cell line was obtained after screening with Hygromycin, and the recombinant retrovirus was packaged, concentrated and purified. The prepared recombinant virus was injected into mice infected with Schistosoma japonicum by intravenous or intraperitoneal route to investigate the expression of the target gene in mice and the effect of in vivo treatment of schistosome infection. Results The target gene was expressed 7 days after the virus was injected into the animal body, and it persisted for more than 45 days. The worm reduction rate of pRevTRE-E77.43-induced mice was 31.0%, which was higher than that of pRevTRE by 1.2% (t = 3.524, P <0.01). The mice induced by pRevTRE-E77.43 had a reduction rate of 35.0% 0.9% higher than pRevTRE (t = 9.485, P <0.01). CONCLUSIONS: pRevTRE-E77.43 can produce a certain degree of therapeutic effect on Schistosoma japonicum infected mice, indicating that the Eustacia philoxeroi E77.43 gene is involved in the biological action of Microtus fortis against Schistosoma japonicum infection.