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目的 探讨幼鼠双侧输尿管梗阻后肾脏 3型钠氢交换蛋白 (Na+ H+ exchangers,NHE)和钠氯协同转运通道蛋白 (thiazide sensitivesodiumcortransporter,TSC)的变化和了解输尿管梗阻解除后利尿现象发生的分子生物学机制。方法 2 4只幼年大白鼠随机分为梗阻组 (BUO ,n =13)及对照组 (n=11) ,梗阻组行双侧输尿管梗阻 2 4h后再解除梗阻 ,对照组仅行输尿管游离而不结扎。每日观察体重、饮水量、食量、尿量变化 ,并留取尿液冰冻待检。 4d后抽取动脉血化验 ,同时取出双侧肾脏 ,右肾全肾快速放入液氮冰冻备份、左肾分离出外髓内带 (ISOM)、外髓外带加肾皮质 (OSOM +C) ,行免疫杂交 (westernblooting)实验 ,检测NHE3和TSC的变化。结果 解除输尿管梗阻后 ,实验组肾脏功能明显受损 ,发生明显的梗阻解除后利尿现象 ,血浆渗透压增高 ,血液中肌苷和尿素氮明显高于对照组 ;外髓内带 (ISOM)和外髓外带加皮质 (OSOM +C)检查 ,发现NHE3均明显降低 (P <0 .0 1和P <0 .0 5 ) ;而TSC在OSOM +C表达也明显下降 (P <0 .0 1)。结论 大白鼠BUO 2 4h肾近曲小管和Henle’s环的粗升管 ,NHE3表达明显下降 ,TSC水平显著降低 ,提示NHE3和TSC减少可能是形成肾小管性钠吸收障碍、尿量增多的主要原因之一。
Objective To investigate the changes of renal sodium exchange protein (NHE) and thiazide sensitive intracortransporter (TSC) after ureteral obstruction in young rats and the molecular biology of diuresis after ureteral obstruction was relieved Learning mechanism. Methods 24 juvenile rats were randomly divided into obstructive group (BUO, n = 13) and control group (n = 11). Obstruction group received bilateral ureteral obstruction for 24 hours and then obstruction was removed. In the control group, only ureter was detached ligation. Daily observation of body weight, water intake, food intake, urine output changes, and to keep urine frozen pending. After 4 days, arterial blood samples were drawn and the bilateral kidneys were removed. The whole kidney of the right kidney was quickly placed in liquid nitrogen for frozen backup. ISOM, OSOM + C, Immunoblotting (westernblooting) experiments, detection of NHE3 and TSC changes. Results After ureteral obstruction was relieved, renal function of the experimental group was significantly impaired, diuresis was obvious after the obstruction was relieved, plasma osmolality increased, blood creatinine and urea nitrogen were significantly higher than those in the control group. Extramedullary extracortical cortex (OSOM + C) examination found that NHE3 were significantly lower (P <0.01 and P <0.05), while the expression of TSC in OSOM + C was also significantly decreased (P <0.01 ). Conclusions The expression of NHE3 and the level of TSC in the proximal tubule and Henle’s ring of rats with BUO 2 4h were significantly decreased, suggesting that the reduction of NHE3 and TSC may be the main reason for the formation of tubular sodium-uptake disorder and increased urine output one.