急性肾衰大鼠肾脏上皮钠通道和钠钾氯共转运蛋白的表达

doi:吉林省科技厅科研基金资助课题 (200705356)

急性肾衰大鼠肾脏上皮钠通道和钠钾氯共转运蛋白的表达

李洪岩¹，高航²，程润芬²，祁霞²，李爽²，王建伟²

1.吉林大学基础医学院病理生理学教研室,吉林长春 130021; 2.吉林大学基础医学院病理生物学教育部重点实验室，吉林长春 130021

收稿日期:2008-09-01 修回日期:1900-01-01 出版日期:2009-01-28 发布日期:2009-01-28
通讯作者: 王建伟

NKCC₂ and γENaC expressions in kidney tissues of rats with acute renal failure

LI Hong-yan¹,GAO Hang²,CHENG Run-fen²,QI Xia²,LI Shuang²,WANG Jian-wei²

1.Department of Pathophysiology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China; 2.Key Laboratory of Pathobiology,Ministry of Education,School of Basic Medical Sciences,Jilin University,Changchun 130021,China

Received:2008-09-01 Revised:1900-01-01 Online:2009-01-28 Published:2009-01-28
Contact: WANG Jian-wei

摘要/Abstract

摘要： 目的：观察急性肾功能衰竭（ARF）时钠转运蛋白NKCC₂和γENaC的水平及细胞定位，探讨ARF时水盐代谢障碍的机制。方法：ARF组大鼠肌注甘油建立ARF模型，对照组(Control)则给等量生理盐水；观察肾组织病理学改变，检测血清及尿液生化指标，应用免疫组化和蛋白印迹方法检测肾脏NKCC₂和γENaC表达及胞内定位。结果：肌注甘油后第1、2和3天 ARF组大鼠尿量分别为（35.2±8.7）、（47.9±11.0）和（43.2±17.0） mL•d^-1，明显高于对照组(P<0.05)，〖JP3〗第4天开始逐渐恢复到对照组水平［(22.0±5.2 )mL•d^-1］；〖JP〗ARF大鼠血肌酐和尿素氮分别为（53.8±12.2）和（9.5±1.2） mmol•L^-1，明显高于对照组［分别为（38.9±9.1 ）和（7.9±1.5） mmol•L^-1］ (P<0.05)；ARF组大鼠肾小管管腔变小，上皮细胞发生脂肪变性，管周有炎细胞浸润；免疫组化检测ARF组大鼠NKCC₂和γENaC表达增加；蛋白印迹光密度比值分析，肾小管上皮细胞膜NKCC₂和γENaC蛋白增加比例(分别为2.54±0.27和1.83±0.23)明显高于细胞质(分别为1.53±0.17和1.21±1.18) (P<0.05)。结论：ARF时细胞膜及细胞质NKCC₂和γENaC表达均增高，以细胞膜增高更为明显，提示AFR时肾组织相对正常肾单位代偿性功能增强，通过钠转运蛋白水平及细胞定位的变化参与水盐代谢的调节。

关键词: 上皮钠通道, 钠钾氯共转运体

Abstract: Abstract:Objective To study the expression and location of Na⁺-K⁺-2Cl^-cotransporter (NKCC₂) and gamma epithelial Na⁺ channel(γENaC) in kidney tissues of rats with acute renal failure (ARF) and to explain the pathophysiological mechanism of water and sodium disorders during ARF.Methods The ARF rats were induced with glycerol by intramuscular injection (ARF group),the control rats were administered with the equal volume of normal saline (control group).Then the pathological alteration of kidney tissues and the serum and urine biochemical parameters of rats were examined,and the expressions of NKCC₂ and γENaC and their cellular location in kidney tissues were also determined by Western blotting and immunohistochemistry (IHC).Results The urine volumes (mL•d^-1)(35.2±8.7,47.9±11.0, and 43.2±17.0 on the first,second and third day, respectively)increased after intramuscular injection of glycerol,they were higher than that of control (P<0.05) ,but decreased gradually to the normal level on the fourth day(22.0±5.2). The serum creatinine and blood urea nitrogen levels in ARF rats were (53.8±12.2) μmol•L^-1 and (9.5±1.2) mmol•L^-1 respectively,and they were higher than those in control group (38.9 μmol•L^-1 ±9.1 μmol•L^-1 and 7.9 mmol•L^-1±1.5 mmol•L^-1 respectively) (P<0.05).　The pathological results showed the considerably shrinked renal tubules in diameter,the adipose degeneration of tubular epithelial cells and the immune cell infiltration in the kidney tissues of ARF rats.The increased NKCC₂ and γENaC expressions of ARF rats were found by IHC,and this was further proved with Western blotting,the ratio of membrane NKCC₂ and γENaC expressions in tubular epithelial cells (2.54±0.27 and 1.83±0.23) increased significantly when compared with the ratio of plasma (1.53±0.17 and 1.21±1.18) (P<0.05).Conclusion During ARF period,both membrane and cytoplasm NKCC₂ and γENaC expressions increase,and the increased membrane protein is the key step for sodium reabsorption.And this suggests that the compensatory enhanced function of the partial nephron with normal structure occurs,and the membrane location or the increased level of sodium transporter involves in the metabolism of sodium and water in the diseased kidney.

Key words: epithelial sodium channel, sodium potassium chloride cotransporter

中图分类号:

R-332

李洪岩，高航，程润芬，祁霞，李爽，王建伟. 急性肾衰大鼠肾脏上皮钠通道和钠钾氯共转运蛋白的表达[J]. J4, 2009, 35(1): 92-95.

LI Hong-yan,GAO Hang,CHENG Run-fen,QI Xia,LI Shuang,WANG Jian-wei. NKCC₂ and γENaC expressions in kidney tissues of rats with acute renal failure[J]. J4, 2009, 35(1): 92-95.