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目的 通过RNA干扰和基因转染技术,研究沉默PAX2基因对UUO大鼠肾间质纤维化的影响.方法 将PAX2-siRNA转染至UUO大鼠肾脏被膜下,将64只幼年雄性大鼠随机分为:阴性对照组(NC)32只;沉默组(RNAi) 32只,于转染后3、5、7、14天分为4组,每组8只,留取肾组织标本,应用Real-Time PCR及Western blot检测肾皮质PAX2 mRNA及其蛋白的沉默情况,HE染色和Masson染色方法在光镜下观察肾小管损伤情况和肾间质纤维化程度.结果 ①沉默组与对照组比较PAX2 mRNA和PAX2蛋白表达量降低,有统计学意义(P＜0.05),②HE和Masson染色观察到对照组随梗阻时间延长,肾小管损伤逐渐明显,肾间质中可见胶原蛋白逐渐增加.沉默组与对照组比较在7d以前无明显变化(P＞0.05);在梗阻14d沉默组与对照组比较肾小管损伤减轻、肾间质病变减低、胶原纤维沉积减少(P＜0.05).结论 在UUO大鼠模型中,梗阻早期沉默PAX2基因对肾间质纤维化进程无明显影响,梗阻晚期可减轻肾小管损伤及肾间质纤维化病变,对肾间质纤维化有明显治疗作用.“,”Objecitve To investigate the effects of the silencing of PAX2 gene on renal interstitial fibrosis in unilateral ureteral obstruction rat model by RNA interference and gene transfection.Methods PAX2 -siRNA was transfected into the renal capsule in unilateral ureteral obstruction rat model;64 young male rats were randomly distributed into negative control group (NC) 32 rats and silencing group (RNAi) 32 rats,and the rats were distributed into 4 groups by 3,5,7,14 days after transfection and 8 rats each groups.Then the renal tissue was taken and the expression of PAX2 mRNA and protein was determined by real-time quantitative polymerase chain reaction (PCR) and Western blot respectively;HE and Masson staining showed the appearance of tubular injury renal and fibrosis after urinary obstruction.Results The expression of PAX2 mRNA and protein of RNAi group was lower than that of NC group,and there were significant differences between NC and RNAi group (P ＜ 0.05).HE and Masson staining showed that nephric tubular affection aggravated and collagen fibers proliferated gradually with prolonged obstruction in control group;There was no obvious changes between silencing group and control group at 3d,5d,7d (P ＞ 0.05),while compared with control group,degrees of tubular injury lessened,pathological changes in renal interstitum degraded and fibrotic proceeding delayed greatly at 14 in silencing group (P ＜ 0.05).Conclusions The silencing of PAX2 has no obvious influence on the progress of renal interstitial fibrosis in the earlier period of obstruction in UUO rat model.Nevertheless,it can lessen the appearance of tubular injury and renal interstitial fibrosis in the advanced stage of renal interstitial fibrosis,therefore providing a new tool to specifically target renal interstitial fibrosis for therapeutics.