吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (4): 834-841.doi: 10.13481/j.1671-587X.20210403

• 基础研究 • 上一篇    下一篇

血管紧张素(1-7)对小鼠肢体缺血再灌注所致肾损伤的改善作用及其机制

朱丽艳1,2,王耀明1,秦政1,赵欢欢1,王增颖1,杨秀红1,2()   

  1. 1.华北理工大学基础医学院机能实验室,河北 唐山 063000
    2.河北省唐山市慢性疾病重点实验室,河北 唐山 063000
  • 收稿日期:2020-11-13 出版日期:2021-07-28 发布日期:2021-07-22
  • 通讯作者: 杨秀红 E-mail:ljkyxhljn@163.com
  • 作者简介:朱丽艳(1975-),女,河北省唐山市人,讲师,医学硕士,主要从事肢体缺血再灌注损伤方面的研究。
  • 基金资助:
    国家自然科学基金项目(81372029);河北省唐山市科技局科技计划项目(14130244B);华北理工大学大学生创新创业训练计划项目(X2017158)

Improvement effect of angiotensin(1-7) on kidney injury induced by limb ischemia-reperfusion in mice and its mechanism

Liyan ZHU1,2,Yaoming WANG1,Zheng QIN1,Huanhuan ZHAO1,Zengying WANG1,Xiuhong YANG1,2()   

  1. 1.Functional Laboratory,School of Basic Medical Sciences,North China University of Science and Technology,Hebei Province,Tangshan 063000,China
    2.Key Laboratory of Chronic Diseases of Tangshan City,Hebei Province,Tangshan 063000,China
  • Received:2020-11-13 Online:2021-07-28 Published:2021-07-22
  • Contact: Xiuhong YANG E-mail:ljkyxhljn@163.com

摘要: 目的

观察血管紧张素(1-7)[Ang-(1-7))]预处理肢体缺血再灌注(LIR)小鼠肾组织和血清中Ang-(1-7)水平及肾组织中核因子κB(NF-κB)、肿瘤坏死因子α(TNF-α)及Mas受体蛋白表达水平和分布,探讨Ang-(1-7)对LIR小鼠肾损伤的影响及其炎症机制。

方法

30只10周龄C57BL/6小鼠随机分为对照组、LIR组和LIR+Ang-(1-7)组,每组10只。采用双后肢止血带套扎阻断血流2 h、再灌注4 h复制小鼠LIR模型。LIR+Ang-(1-7)组小鼠于造模前14 d埋置渗透泵皮下给予Ang-(1-7)(24 μg·kg-1·h-1),LIR组小鼠于造模前14 d埋置渗透泵皮下给予等量生理盐水。采用全自动生化分析仪检测小鼠血清中血肌酐(SCr)和尿素氮(BUN)水平,采用HE染色观察小鼠肾组织病理形态表现并进行肾组织病理损伤评分,采用酶联免疫吸附测定法(ELISA)检测小鼠肾组织和血清中Ang-(1-7)水平,免疫组织化学染色法和Western blotting法检测小鼠肾组织中NF-κB、TNF-α和Mas受体蛋白表达分布情况和表达水平。

结果

与对照组比较, LIR组小鼠血清中SCr和BUN水平明显升高(P<0.05),肾组织有明显充血、肾小管扩张、水肿、坏死和炎细胞浸润等病理损害,肾组织病理损伤评分明显升高(P<0.05),肾组织和血清中Ang-(1-7)水平明显降低(P<0.05),肾组织中NF-κB、TNF-α和Mas受体蛋白表达水平明显升高(P<0.05);与LIR组比较,LIR+Ang-(1-7)组小鼠血清中SCr和BUN水平明显降低(P<0.05),肾组织充血等病理损害明显减轻,肾组织病理损伤评分明显降低(P<0.05),肾组织及血清中Ang-(1-7)水平明显升高(P<0.05),肾组织中NF-κB和TNF-α蛋白表达水平明显降低(P<0.05),Mas受体蛋白表达水平明显升高(P<0.05)。

结论

小鼠LIR可诱导远端肾脏损伤,Ang-(1-7)可能通过降低肾组织中NF-κB和TNF-α蛋白表达水平进而对小鼠LIR所致肾损伤起到改善作用。

关键词: 缺血再灌注, 急性肾损伤, 肾素-血管紧张素系统, 炎症

Abstract: Objective

To explore the effect of angiotensin (1-7) [Ang-(1-7)] on renal injury and inflammation in the limb ischemia-reperfusion (LIR) mice by detecting the level of Ang-(1-7) in kidey tissue and serum and the expression levels of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α) and Mas receptor protein in kindey tissue of the LIR mice pretreated with Ang-(1-7).

Methods

Thirty 10-week-old C57BL/6 mice were randomly divided into control group, LIR group, and LIR+Ang-(1-7) group, with 10 mice in each group. The mice in LIR group and LIR+Ang-(1-7) group were subjected to 2 h of ischemia and 4 h of reperfusion to establish the mouse LIR models. The mice in LIR+Ang-(1-7) group were treated Ang-(1-7) (24 μg·kg-1·h-1) for two weeks before LIR by placing subcutaneous osmotic pump. The mice in LIR group were treated the same amount of normal saline for two weeks before LIR by placed subcutaneous osmotic pump. Automatic biochemical analyzer was used to determine the levels of serum creatinine (SCr) and urea nitrogen (BUN) of the mice; HE staining was used to observe the pathomorphology of kidney tissue of the mice, and renal tubular pathological score was performed. Enzyme linked immunosorbent assay (ELISA) was used to determine the Ang-(1-7) levels in kidney tissue and serum of the mice. Immunohistochemical staining and Western blotting methods were used to detect the expression, distribution and the expression levels of NF-κB,TNF-α and Mas receptor proteins in the mouse kidney tissue.

Results

Compared with control group, the levels of serum SCr and BUN of the mice in LIR group were increased significantly (P<0.05), and the obvious pathological damages such as hyperemia, renal tubular dilation, edema, necrosis, and inflammatory cell infiltration were observed in the kidney tissue; the pathological injury score of kidney tissue was increased(P<0.05); the Ang-(1-7) levels in kidney tissue and serum of the mice were decreased significantly (P<0.05), and the expression levels of NF-κB, TNF-α and Mas receptor proteins were increased significantly (P<0.05). Compared with LIR group, the levels of serum SCr and BUN of the mice in LIR+Ang-(1-7) group were decreased significantly (P<0.05),and the kidney congestion and other damages were significantly reduced;the pathological injury score of kindey tissue was decreased(P<0.05); the Ang-(1-7) levels in kidney tissue and serum of the mice were increased significantly (P<0.05); the expression levels of NF-κB and TNF-α proteins were decreased (P<0.05), and the expression level of Mas receptor protein in kidney tissue of the mice in LIR+Ang-(1-7) group was increased significantly (P<0.05).

Conclusion

Mouse LIR can induce the distal kidney damage, and Ang-(1-7) may reduce the expression levels of NF-κB and TNF-α proteins in kidney tissue to improve the kidney damage caused by mouse LIR.

Key words: ischemia-reperfusion, acute kidney injury, renin-angiotensin system, inflammation

中图分类号: 

  • R363