巨细胞病毒感染对细胞肾素基因表达的影响及其生物学意义

Cytomegalovirus infects its host cells and induces their renin gene expression

目的探讨巨细胞病毒（cytomegalovirus，CMV）感染肾球旁细胞肾素基因表达的变化及其意义。方法用病毒感染复数（MOI）为10、0．1和0的鼠CMV分别与鼠肾球旁细胞模型As4．1细胞共育5d作为实验组；用紫外线灭活病毒的假感染（mock感染）对照组。RT—PCR检测感染细胞中CMV即刻早期基因1（IE1）的表达；免疫荧光观察肾素阳性细胞和肾素阳性颗粒在细胞的分布；双色免疫荧光染色观察肾素阳性颗粒是否出现在CMV阳性细胞；RT—PCR和Westernblot检测。肾素基因在感染细胞内的表达。结果CMV感染As4．1细胞后出现典型的病毒空斑；病毒感染细胞CMV IE RT—PCR产物阳性；肾素阳性细胞集中在病毒空斑周围CMV新感染细胞区，肾素阳性荧光颗粒主要以块状和环状存在于病毒感染细胞质中；双色免疫荧光染色显示肾素阳性颗粒和CMV阳性颗粒出现在同一细胞；CMV感染细胞肾素基因的表达随病毒感染量增加而增加。结论CMV感染并导致其宿主细胞肾素基因表达，可能涉及CMV加速心血管疾病发生发展的新机制。

Objective To explore renin expression in cytomegalovirus（CMV） infected juxtaglomerular cells（JG） and its biological significance, Methods JG model cell line As4.1 cells derived from kidney tissue were respectively incubated with murine CMV at multiplicities of infection （MOI） of 10, 0.1 and 0 for 5 d, and the control was mock infection with the same amount of ultraviolet inactive CMV as MOI 10, then the cells were harvested. CMV immediate early gene（ IE1 ） mRNA in the cells was tested by RT-PCR. The renin positive cells and the renin fluorescence granules in the cells were examined by immunofluorescence stain. Whether or not renin antigen and CMV antigen were showed in the same cells by FITC and TRITC immunofluorescence. The renin gene expression in the cells was individually detected by real-time RT-PCR and Western blot. Results The ceils infected by CMV showed typical cytopathic effect（CPE） and viral plaques in the cell monolayer. CMV IE1 mRNA was found in the viral infected cells by RT-PCR. The mass or ring granules of renin positive fluorescence appeared in the cytoplasm of the CPE cells. The renin positive cells congregated around the viral plaques. Renin positive granules and CMV positive granules showed in the same cells. Renin expression in the CMV infected cells exhibited in a dependent manner of ratio of infectious virus particles to cells. Conclusion CMV infection of the cells derived from kidney tissue induces renin expression related to a new pathogenesis of cardiovascular diseases.