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目的 研究活体大鼠肾脏中慢病毒载体介导的基因转染及表达情况。方法 以水泡性口炎病毒包膜蛋白 (VSV- G)为包膜、携带以磷酸甘油酸激酶 (PGK)启动子启动的 L ac Z报告基因的慢病毒载体注射 L ewis大鼠右侧肾脏实质 ,分别于注射后 1、2、3、7天处死大鼠 (每个时间点实验组 n=3,对照组 n=2 ) ,通过β-半乳糖苷酶(β- Gal)组织化学染色检测报告基因的表达。结果 实验组大鼠右肾可见 β- Gal染色阳性细胞 ,而对侧肾脏及其他脏器均未见 β- Gal表达 ;β- Gal的表达在肾实质注射 2天后达到峰值 ,到第 7天仍能维持较高水平。β- Gal阳性细胞周围未见明显炎症细胞浸润。病毒载体注射的肾脏外观及组织形态学未见异常。结论 慢病毒载体能有效而稳定的将外源基因导入活体大鼠肾脏而无明显毒副作用
Objective To study the lentiviral vector-mediated gene transfection and expression in living rat kidney. Methods The right renal parenchyma of L ewis rats were injected with the lentiviral vector carrying the L ac Z reporter gene activated by the phosphoglycerate kinase (PGK) promoter and encapsulated by vesicular stomatitis virus envelope protein (VSV-G) The rats were sacrificed at 1, 2, 3 and 7 days after injection (n = 3 in the experimental group at each time point and n = 2 in the control group), and detected by β-galactosidase Reporter gene expression. Results The β-Gal staining positive cells were found in the right kidney of the experimental group, while the expression of β-Gal was not seen in the contralateral kidney and other organs. The expression of β-Gal peaked at 2 days after injection of renal parenchyma, Can maintain a high level. There was no obvious inflammatory cell infiltration around the β-Gal positive cells. The appearance and histomorphology of kidney injected with viral vector showed no abnormalities. Conclusion The lentiviral vector can effectively and stably transfer the exogenous gene into the living rat’s kidney without any obvious toxic or side effects