急性肾损伤时水通道蛋白2及钠钾泵的表达

The expression of water channel protein 2 and Na^＋-K^＋-ATPase in acute kidney injury

目的 探讨水通道蛋白2的表达及钠-钾-三磷酸腺苷酶在多器官功能障碍肾损伤发病机制中的作用,研究大鼠多器官功能障碍肾损伤与水通道蛋白2及钠-钾-三磷酸腺苷酶的表达.方法 选取健康大鼠72只,随机（随机数字法）分为对照组24只,脂多糖组48只,采用大鼠腹腔注射脂多糖5 mg/kg造成内毒素致多器官功能障碍动物模型,对照组仅作假手术处理.造模成功后6h、12h、24 h、2d、3d、5d分别处死动物,留取血液、尿量进行生化检测,并应用免疫组化法和RT-PCR技术检测肾组织内水通道蛋白2 mRNA及蛋白的表达水平.应用试剂盒测定钠-钾-三磷酸腺苷酶的含量及活性.结果 造模成功后致伤组大鼠尿量迅速减少,48 h后尿量增多.尿素氮、肌酐逐渐增高,48 h达高峰,此后逐渐下降.水通道蛋白2 mRNA及蛋白表达迅速减少,48 h降至最低,此后逐渐增多.钠-钾-三磷酸腺苷酶含量差异无统计学意义,但其活性在造模成功后明显降低,此后逐渐增高,但仍低于对照组.结论 大鼠多器官功能障碍综合征肾损伤模型中,水通道蛋白2是肾脏重吸收功能恢复的结构基础,钠-钾-三磷酸腺苷酶则直接参与或间接反映肾脏的能量代谢状态,只有在水通道蛋白2及能量代谢恢复后,大鼠的肾脏功能才能逐渐好转.

Objective To investigate the role of AQP2 and Na ＋-K ＋-ATPase in the pathogenesis of kidney injury with multiple organ dysfunction syndrome,and try to find the express characteristics of them.Methods A total of 72 healthy rats were randomly （random number） divided into two groups：control group （n =24） and Lipopolysaccharide group （n =48）.The Lipopolysaccharide group rats were injected with 5 mg/kg lipopolysaccharide at the beginning while the control group was 0.9％ sodium chloride.After the model was succeeded,the rats were put to death at 6 h,12 h,24 h,2 days,3 days and 5 days equally.The urine and blood were collected.Blood were used biochemical tests to check.kidney AQP2 protein and mRNA expression level in the organization were applied the immune organized and RT-PCR technique to detect.Applied kit for determining the content and activity of sodium/potassium-atpase.Results The volume of urine in LPS group decrease quickly at 12 h and 24 h,but increased after 2 days.Urea nitrogen and creatinine increased gradually,and peaked at 48 h,after then gradually decline.AQP2 mRNA and protein expression decreased,and minimize at 48 h.The content of Na＋-K＋-ATP ase has no obvious difference,but the activity significantly decreased at the beginning,then increased gradually,but it was still lower than the control group.Conclusions In renal injury rats model with multiple organ dysfunction syndrome,AQP2 is the structure of renal reabsorption function,while Na ＋-K ＋-ATPase directly involved in or indirectly reflected the state of kidney energy metabolism.Recovery of AQP2 protein and energy metabolism,before the rat kidney function improved.