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目的探讨巨细胞病毒(Cytomegalovirus,CMV)是否感染血管内皮细胞伴肾素表达。方法(1)用107pfu(空斑形成单位)/ml CMV临床分离株BI-5和实验室型CMV AD169分别与106腹主动脉内皮细胞和人脐静脉内皮细胞共同孵育,在23小时后、第3天、第7天、第10天和第14天分别收集培养上清200μl,第14天用PBS缓冲液洗细胞3次,收获细胞。每组实验均设培养液代替病毒液的无感染对照;(2)COBAS定量PCR检测培养上清中CMV DNA拷贝数;(3)PCR检测感染细胞中CMVpol基因;(4)RT-PCR、Real time RT-PCR和Western blot检测肾素在感染细胞内的表达。结果(1)BI-5和AD169感染静脉和动脉细胞后,其形态学变化相似,无细胞裂解病理效应;(2)AD169感染细胞不同时间培养上清中CMV DNA拷贝数无明显增加,BI-5呈增殖趋势;(3)BI-5感染动脉细胞CMV DNA拷贝数和肾素表达量均大于静脉细胞。结论临床分离株CMV以非裂解形式在血管内皮细胞持续存在并诱导肾素基因表达,血管内皮细胞分泌肾素可能是CMV感染引起心血管疾病的新机制。
Objective To investigate whether Cytomegalovirus (CMV) infects vascular endothelial cells with renin expression. Methods (1) 107pfu (plaque forming unit) / ml CMV clinical isolates BI-5 and laboratory-type CMV AD169 were co-incubated with 106 abdominal aortic endothelial cells and human umbilical vein endothelial cells, respectively. After 23 hours, On the 3rd, the seventh, the tenth and the fourteenth days, 200 μl of the culture supernatant was collected, and the cells were washed three times with PBS buffer on the 14th day. (2) COBAS quantitative PCR detection of CMV DNA copy number in the culture supernatant; (3) PCR detection CMVpol gene in infected cells; (4) RT-PCR, Real The expression of renin in infected cells was detected by RT-PCR and Western blot. Results (1) The morphological changes of BI-5 and AD169 after infection of vein and arterial cells were similar and there was no pathological effect of cell lysis. (2) There was no significant increase of copy number of CMV DNA in the culture supernatant of AD169 infected cells, 5 showed a trend of proliferation. (3) CMV DNA copy number and renin expression in BI-5-infected arterial cells were larger than that of vein cells. Conclusion The clinical isolates CMV persist in vascular endothelial cells in non-lytic form and induce renin gene expression. The secretion of renin by vascular endothelial cells may be a new mechanism of CMV-induced cardiovascular disease.