缺血再灌注损伤后大鼠肾小管上皮细胞有机阴离子转运蛋白的表达及其功能  

Expression and function of organic anion transporters after ischemia reperfusion injury in rat kidney tubular epithelial cells

作　　者：张锐[1] 阳晓[1] 李军[1] 伍军[1] 彭文兴[1] 董秀清[1] 余学清[1] 

机构地区：[1]中山大学附属第一医院肾内科卫生部肾脏病临床研究重点实验室,广州510080

出　　处：《中华肾脏病杂志》2008年第2期119-124,共6页Chinese Journal of Nephrology

基　　金：广东省中医药管理局科研基金（1060173）

摘　　要：目的探讨大鼠肾脏缺血再灌注损伤（IRI）后肾小管上皮细胞有机阴离子转运蛋白（OAT）1和3表达及功能变化。方法建立大鼠IRI模型，于术后第1、2、4、6天取血及肾组织标本。常规测定Scr浓度及观察肾脏组织形态。分别用免疫组织化学和Western印迹检测肾脏组织OAT1和3蛋白表达。毛细血管电泳仪检测肾脏对氨基马尿酸（PAH）排泄率。钼蓝分光光度法测定基侧膜钠-钾-ATP酶（Na^＋-K^＋-ATP酶）活性。另设假手术组作对照。结果模型组大鼠术后第1天肾小管上皮细胞坏死脱落，管腔内可见大量管型，刷状缘消失，基底膜裸露；Scr显著升高。与假手术组比较，模型组大鼠肾组织匀浆及肾小管上皮细胞OAT1和OAT3表达增强；PAH排泄率降低[（0．53±0．15）比（4．00±0．67）ml／min，P〈0．01]；肾小管上皮细胞基侧膜Na^＋-K^＋-ATP酶活性降低[（9．33±1．37）比（15．07±0．15）μmolPi·（mg蛋白）^-1·h^-1，P〈0．01]。损伤后第2天及第4天，损伤的肾小管逐渐修复，OAT1和3表达逐渐降低，PAH排泄率和Na^＋-K^＋-ATP酶活性逐渐增加；至第6天时肾小管基本恢复正常。结论急性肾缺血再灌注导致大鼠肾小管上皮细胞可逆性损伤，损伤初期肾小管OAT1和OAT3表达增强，转运有机阴离子功能下降，PAH排泄减少。Objective To investigate the expression and function transporter（OAT） 1 and 3 after ischemia reperfusion injury（IRI） in rat kidney of organic anion Methods Rat renal ischemia reperfusion injury models were established and were sacrificed at day 1, 2, 4, 6 after injury. Renal OAT1 as well as OAT3 expression were analyzed by immunohistoehemistry and Western blotting. P-aminohippurate （PAH） excretion was detected by capillary electrophoresis and net secretion of PAH was calculated. Results IRI operation resulted in severe renal tubules damage on day 1 with significantly increased serum creatinine level [（278.9±51.8） vs （41.4±4.6） μmolL, P〈0.01]. Compared with sham-operated rats, there was a significant increment in the expression of OAT1 and OAT3 in renal cortex homogenates and basolateral membranes on day 1 after IRI. While on day 2 and 4 after IRI, the expression of OAT1 and OAT3 in renal cortex homogenates and basolateral membranes decreased gradually but were still significantly higher than that of the sham-operated rats. The excretion rate of PAH was significantly decreased on day 1 compared with that of sham-operatod rats [（0.53±0.15） vs （4.00±0.67） ml/min, P〈0.01] and returned to normal when the tubular injury recovered. Renal tubular basolateral membrane Na^＋-K^＋-ATPase activity decreased significantly on day 1 [（9.33±1.37） vs （15.07±0.15） μmol Pi·（mg pro）^-1·h^-1, P〈0.01]but gradually increased on day 2, 4 and 6 in IRI rats. On day 6 after IRI, renal tubular injury recovered to normal state..Consistently, the Na^＋-K^＋-ATPase activity was regulated to the level comparable to that in sham-operated rats. Conclusion IRI is reversible and the expression levels of OAT1 and OAT3 are elevated in the early stage of IRI while OAT-mediated PAH excretion is markedly decreased, which may be due to diminished activity of the Na^＋-K^＋-ATPase.

关 键 词：有机阴离子转运子 再灌注损伤 肾小管 对氨基马尿酸 

分 类 号：R285.5[医药卫生—中药学]