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小鼠肢体缺血再灌注后肾组织AT1和Mas受体蛋白差异性表达与肾损伤的关系
作者:王建辉 张伟 刘燕 王丽君 王丽萍 范素静 杨秀红 
单位:063000 河北唐山 华北理工大学基础医学院唐山市慢性病临床基础重点实验室(王建辉、张伟、刘燕、王丽君、王丽萍、范素静、杨秀红) 
关键词:再灌注损伤 急性肾损伤 肾素-血管紧张素系统 
分类号:R363
出版年,卷(期):页码:2016,41(3):184-188
摘要:
[摘要] 目的 通过观察小鼠肢体缺血再灌注(LIR)损伤后的肾组织中AT1Mas受体蛋白的表达变化与肾损伤程度的改变,探讨局部组织肾素血管紧张素系统(RAS)稳态失衡在LIR后肾损伤中的作用。方法 止血带套扎双后肢阻断血流,2h后解套扎进行再灌注复制小鼠LIR模型。将48只雄性8周龄ICR小鼠随机分为8组,每组6只,其中一组作为对照组,其余7组为再灌注10min0.5h1h2h4h6h12h模型组。肾组织病理切片常规HE染色观察肾组织形态变化并进行病理损伤评分。全自动生化分析仪测定血清肌酐和尿素的含量。Western blotting检测肾组织AT1Mas受体蛋白的表达变化。结果 病理组织学检测结果显示,LIR后不同时间点小鼠肾组织有充血、水肿、炎细胞浸润和上皮细胞变性等不同程度损伤,且随着灌注时间的延长,损伤评分逐渐升高。生化检测结果显示,肌酐和尿素的含量随灌注时间的延长而增加,再灌注4h达最高。Western blotting检测结果显示,AT1受体蛋白的表达随灌注时间的延长而降低,Mas受体蛋白的表达随灌注时间的延长而增加。结论 LIR后肾组织损伤逐渐加重,AT1/Mas受体蛋白表达失衡可能参与小鼠LIR后肾损伤的发生。

[Abstract]  Objective  To explore the role of imbalance of local renin angiotensin system (RAS) in renal injury by observing the changes of AT1 and Mas receptor protein expression in renal and the degree of renal injury after limb ischemia-reperfusion (LIR) in mice. Methods  The hind legs of ICR mice were rendered ischemic by using a tourniquet to block blood flow. The tourniquet was release after 2 hours to initiate reperfusion. Forty-eight 8-week-old male ICR mice were randomly assigned into 8 groups (6 in each group), including a control group and 7 model groups of 10min, 0.5h, 1h, 2h, 4h, 6h and 12h after reperfusion. Histopathological technique was used to observe morphological changes in renal tissue, and pathological score was assessed. Automatic biochemical analyzer was employed to determine the contents of serum creatinine and urea. Western blotting was performed to determine the expression of AT1 and Mas receptor protein in renal tissue. Results  Pathological changes were found in renal tissue of model mice, including interstitial congestion, edema, inflammatory cell infiltration, and epithelial cell degeneration, and the injury score elevated gradually along with the length of reperfusion time. Biochemical results showed that the contents of serum creatinine and urea increased with the prolongation of reperfusion time, reaching the highest level at 4h point. Western blotting showed that the expression of AT1 protein decreased significantly and of Mas receptor protein increased significantly with the prolongation of perfusion time. Conclusion  The renal damage is gradually increased after LIR, and the expression imbalance of AT1/Mas receptor proteins may be involved in the damage process.

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