【摘要】 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更多还原

【Abstract】 Objective To analyze gene transduction of rat kidney in vivo after administration of lentiviral vectors.Methods The vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped lentiviral vector harboring the LacZ reportor gene driven by phosphoglycerate kinase (PGK) promoter or saline solution (as control) were delivered into the right kidneys of Lewis rats by direct injection into the renal parenchyma.Animals were killed at 1,2,3 and 7 days after treatment(3 cases each time point for experimental animals,2 cases each time point for controls).The transducted gene expression was determined by histochemical staining of β-galactosidase (β-Gal).Results β-Gal staining positive cells were only detectable in the kidneys injected with the lentiviral vector but not in the contralateral kidneys or the other organs.Transgene expression was mostly localised to cortex and corticomedullary junction.Expression level of β-Gal peaked at 2 days after lentiviral transduction and persisted for the entire duration of the experiments.No apparent inflammatory infiltration was observed around β-Gal positive cells.All the lentivirus-injected kidneys demonstrated normal appearance and histology.Conclusion Lentiviral vector can effectively and stably transfer exogenous gene to rat kidney in vivo without significant toxic effects.更多还原