促红细胞生成素对输尿管梗阻解除后幼鼠肾脏水通道蛋白2表达及功能的影响  

Effect of erythropoietin on the expression and function of renal aquaporin -2 after release of bilateral ureter obstruction in young rats

作　　者：王焱[1] 任川川[1] 杨黎[2] 吕宇涛[1] 文建国[1] 黄书满[1] 谢佳丰 贾智明 王庆伟[1] 

机构地区：[1]郑州大学第一附属医院泌尿外科&小儿尿动力中心,450052 [2]郑州大学第一附属医院生物治疗中心

出　　处：《中华实用儿科临床杂志》2016年第5期367-370,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81370869）;郑州市科技局资助项目（131PLJRC657）

摘　　要：目的探讨促红细胞生成素（EPO）对输尿管梗阻-解除梗阻后（BUO—R）幼鼠肾脏水通道蛋白2（AQP2）表达及功能的影响。方法幼年SD大鼠32只，按随机数字表法随机均分为BUO组、BUO．R组、BUO-R＋EPO组和假手术（Sham）组，每组8只。实验组大鼠均行双侧输尿管结扎，BUO组梗阻24h后处死；BUO—R组和BUO-R＋EPO组梗阻24h后解除梗阻，BUO-R＋EPO组分别于解除梗阻后2h、6h、12h、24h和36h腹腔注射EPO（每次剂量500U／kg），48h后处死；BUO—R组在相同时间点腹腔注射等量9g／L盐水，48h后处死；Sham组只分离输尿管，不结扎，72h后处死。幼鼠处死前，代谢笼内收集尿液进行渗透压检测。留取双侧肾脏标本，采用Real—timePCR、免疫组织化学及Westernblot检测各组肾脏组织中AQP2mRNA及蛋白表达水平。结果Sham组尿液渗透压最高，BUO—R＋EPO组次之，BUO—R组最低（P〈0．05）。免疫组织化学结果显示BUO组、BUO—R组和BUO—R＋EPO组集合管管壁变薄、管腔增大，经Image—ProPlus图像分析软件统计分析，BUO组内髓集合管AQP2阳性染色弱于其他3组，Sham组表达最强，BUO—R＋EPO组和BUO—R组次之（P〈0．05）。经Westernblot进一步验证，BUO组AQP2蛋白表达水平低于BUO—R＋EPO组、BUO—R组和Sham组（P〈0．05）。经Real—timePCR检测，4组幼鼠肾脏组织中AQP2mRNA表达水平差异有统计学意义（P〈0．05），Sham组表达最多，是BUO组的（24．30±1．03）倍、BUO—R组的（10．60±1．05）倍、BUO—R＋EPO组的（5．70±1．01）倍。结论EPO可促进BUO—R幼鼠肾脏AQP2mRNA、蛋白表达及功能恢复。Objective To investigate the effect of erythropoietin （EPO） on the expression of aquaporin -2 （AQP2） in the kidney of young SD rats after release of bilateral ureter obstruction （ BUO - R）. Methods Thirty - two young SD rats were equally divided into 4 groups randomly （ BUO group, BUO - R group, BUO - R ＋ EPO group and Sham group,8 rats in each group）. The BUO model was built through bilateral ureteral ligation. EPO （500 U/kg） was given to BUO - R ＋ EPO rats at 2 h after release of BUO, and then repeated 6 h, 12 h,24 h and 36 h thereafter and the same volume of 9 g/L saline was simultaneously given to BUO - R rats. The Sham group was prepared in parallel by laparotomy and free dissection of bilateral ureters but not ligated. Both side kidneys were harvested 48 h （72 h for Sham group） after release of BUO to examine the effect of EPO on the expression of AQP2 in inner medulla by immunohisto- chemistry, Real - time PCR and Western blot. The urine samples were collected by using metabolic cage before death. Results The osmotic pressure of BUO - R ＋ EPO group was higher than that of BUO - R group,but lower than that of Sham group （P 〈 0.05）. Immunohistochemistry showed that the collecting duct wall thinned and lumen enlarged. After the pictures were analysized by using Image - Pro Plus software, it showed that the expression of AQP2 in collecting duct in BUO group was significantly down - regulated compared with that in Sham group, whereas, it was slightly weaker in BUO - R group and BUO - R ＋ EPO group than Sham group（P 〈0.05）. These results were further confirmed by a- dopting Western blot, and the relative quantity of AQP2 in BUO group was also the lowest of the four groups （ P 〈 0.05）. Real - time PCR showed that the level of AQP2 mRNA in Sham group was （24.30±1.03 ） folds of BUO group, （ 10.60 ±1.05 ） folds of BUO - R group and （ 5.70 ± 1.01 ） folds of BUO - R ＋ EPO group, respectively. Conclusion EPO could promote not only the recovery of AQP2 m

关 键 词：促红细胞生成素 输尿管梗阻 水通道蛋白2 肾脏 

分 类 号：R726.9[医药卫生—儿科]