肾脏缺血再灌注损伤E-钙粘附素的表达研究  

The change of E-cadherin expression in renal ischemia reperfusion injury

作　　者：刘书馨[1] 陈香美[1] 孙雪峰[1] 尹忠[1] 吕杨[1] 师锁柱[1] 洪权[1] 刘维萍[1] 

机构地区：[1]解放军总医院肾内科解放军肾脏病研究所暨肾病重点实验室,北京100853

出　　处：《中国实用内科杂志》2007年第6期432-434,共3页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金创新研究群体科学基金项目(30121005)

摘　　要：目的探讨肾脏缺血再灌注损伤时E-钙粘附素(E-cadherin)的变化。方法2006年1月至6月对解放军肾脏病研究所暨肾病重点实验室利用ATP耗竭剂抗霉素A建立的犬集合管上皮细胞(MDCK)缺血模型进行体外实验;体内实验使用双侧肾动脉钳闭法建立大鼠缺血再灌注损伤模型。PAS染色了解肾脏病理改变;E-cadherin免疫组化了解E-cadherin的定位和表达;Western bolt检测E-cadherin的蛋白表达。结果正常MDCK细胞E-cadherin在细胞膜表达,ATP耗竭4hE-cadherin表达极性消失,主要在细胞质表达。肾组织免疫组化显示,假手术组E-cadherin主要表达在肾小管上皮细胞基底膜侧;缺血肾脏组E-cadherin的表达极性消失,空泡变性的肾小管上皮细胞基底膜侧E-cadherin表达明显减少,主要在细胞质表达;而在脱落的肾小管上皮细胞只表达于细胞质。肾组织Western bolt显示,假手术组E-cadherin为120ku条带,缺血45min120ku条带减弱,出现明显的80ku条带,缺血60min120ku条带几乎检测不出,而主要表现为80ku条带。结论体内体外实验均证实,肾脏缺血再灌注损伤时E-cadherin的定位、蛋白表达量发生明显变化,并出现时间依赖性E-cadherin降解,使上皮细胞的极性和细胞间连接受损,部分解释了肾脏缺血再灌注损伤时上皮细胞脱落的原因。Objective To explore the changes of E-cadherin expression in renal ischemia-reperfusion injury. Methods For the in vitro analysis of epithelial ischemia, confluent monolayers of MDCK cells growing in DMEM were depleted of ATP for 4 h by incubation in PBS （ supplemented with 1.5 mM CaCl2 and 2 mM MgCl2 ） containing 10 μM antimycinA, For the in vivo studies of epithelial ischemic injury, adult Sprague-Dawley rats were subjected to bilateral renal artery ligation. Renal pathological changes were measured by PAS stain. Lecadon and expression of E-cadherin were detected by immunohistochemistry and western blot respectively. Results E-cadherin were primarily found in a linear pattern at the lateral portions of the plasma membrane in normal MDCK. After ATP depletion for 4 hours, the linear pattern altered and manifested by the appearance of intracellular staining. In invivo ischemia-reperfusion model rats, E-cadherin expression was changed from normal tubular epithelial cell basal membrane to cytoplasma. Western blot suggested that in sham-operated rats,E-cadherin was 120 ku lane vs 80 ku lane in ischemia for 60 min rats,while in ischemia for 45 min rats,both the 120 ku and 80 ku lanes were detected. Conclusion In renal ischemia-reperfusion,the location and expression of E-cadherin are obviously altered in vivo and in vitro study and E-cadherin are degradated as ischemia time prolongs. These changes may be the reason why tubular epithelial cell exfoliated from TBM in ischemia-reperfusion injury.

关 键 词：E-钙粘附素 肾脏 缺血再灌注损伤 细胞极性 

分 类 号：R5[医药卫生—内科学]