促红细胞生成素对单侧输尿管部分梗阻幼鼠肾脏水通道蛋白2表达的影响  

Effect of erythropoietin on expression of aquaporin-2 in kidney of young rats with partial unilateral ureter obstruction

作　　者：文建国[1] 任川川[1] 吕宇涛[1] 邢璐 杨黎 李真珍 王焱 苏志强 夏明亮 王庆伟[1,3] 程敬亮[2] 

机构地区：[1]郑州大学第一附属医院小儿尿动力中心和泌尿外科,450052 [2]郑州大学第一附属医院磁共振科,450052 [3]河南省高等学校临床医学重点学科开放实验室

出　　处：《中华实用儿科临床杂志》2013年第5期337-340,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（30571931）

摘　　要：目的探讨促红细胞生成素（EPO）对单侧输尿管部分梗阻（PUUO）幼鼠。肾脏水通道蛋白2（AQP2）表达的影响。方法SD幼鼠24只，随机分为PUUO组、PUUO＋EPO组和假手术组，各8只。PUUO组和PUUO＋EPO组幼鼠采用输尿管腰大肌埋植法建立左侧输尿管部分梗阻模型，PUUO＋EPO组术后给予EPO腹腔注射（500U／kg），首次于术后1h给予，其余隔天1次，共4次；PUUO组给予同等剂量9g／L盐水。假手术组仅分离输尿管，不包埋。术后1周处死大鼠，取左侧’肾脏标本，采用荧光定量PCR、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。结果免疫组织化学结果显示PUUO组内髓集合管AQP2阳性染色弱于PUUO＋EPO组和假手术组，经Westernblot进一步验证，PUUO组AQP2蛋白表达水平低于PUUO＋EPO组和假手术组（分别为0．290±0．016、0．640±0．106和0．840±0．134，P〈0．05）。经荧光定量PCR检测，3组幼鼠肾脏组织中AQP2mRNA表达水平差异有统计学意义，假手术组表达最多，PUUO＋EPO组次之，PUUO组表达最少（3组ACt值分别为-7．60±0．18、-9．70±0．54和-10．80±0．31，P〈0．05）。结论EPO有阻止PUUO幼鼠肾脏AQP2mRNA及蛋白表达下调的功能。Objective To investigate the effect of erythropoietin （EPO） on expression of aquaporin-2 （AQP2） in the kidney of young SD rats with partial unilateral ureter obstruction （PUUO）. Methods Twenty-four young SD rats were randomly divided into 3 groups （ PUUO group, n = 8 ; PUUO ＋ EPO group, n = 8 ; sham group, n = 8）. The PUUO model was built by embedding the upper quarter of the left ureter into psoas muscle. EPO （500 U/kg） was given to PUUO ＋ EPO rats at 1 h after surgery,and then repeated 1,3,5 days later after surgery by intraperitoneal injection and the same volume of 9 g/L saline was simuhaneously given to PUUO rats. The sham group was prepared in parallel by way of laparotomy and free dissection of the left ureter. In 1 week after surgery, the left kidneys of rats were harvested to examine the effect of EPO on the expression of AQP2 in inner medulla by immunohistochemistry, real-time PCR and Western blot. Results Immunohistochemistry showed that the expression of AQP2 in the collecting duct in PUUO group was significantly downregulated than those in sham group and PUUO ＋ EPO group, whereas,it was slightly weaker in PUUO ＋ EPO group than those in sham group. These results were further confirmed by Western blot, and the relative quantity of AQP2 in PUUO group was also lower than those in PUUO ＋ EPO group and sham group （0. 290 ± 0. 016 vs 0. 640 ± 0. 106 vs 0. 840 ± 0. 134 ,P 〈 0.05 ）. Real-time PCR showed that the level of AQP2 mRNA in PUUO group was lower than those in PUUO ＋ EPO group and sham group （ ACt values of 3 groups were - 7.60 ± 0. 18 vs - 9.70 ± 0.54 vs - 10.80± 0.31, P 〈 0.05 ）. Conclusions EPO can partially prevent the downregulation of AQP2 in young PUUO rats, and its mechanism needs to be further investigated.

关 键 词：促红细胞生成素 单侧输尿管部分梗阻 水通道蛋白2 肾脏 

分 类 号：R726[医药卫生—儿科] R6[医药卫生—临床医学]