补体C5a/C5aR通路在小鼠缺血/再灌注肾损伤中的作用及机制  

Effects and mechanism of complement C5a/C5aR pathway on ischemiareperfusion induced acute kidney injury in mice

作　　者：张坤 殷淑君 张延[1] 郑权友(指导) ZHANG Kun;YIN Shujun;ZHANG Yan;ZHENG Quanyou(Department of Urology,the 305th Hospital of PLA,Beijing 100017,China)

机构地区：[1]解放军第305医院泌尿外科,北京100017 [2]北京回龙观医院检验科,北京100096 [3]陆军军医大学西南医院第958医院泌尿科,重庆400020

出　　处：《中国免疫学杂志》2022年第22期2700-2705,共6页Chinese Journal of Immunology

基　　金：后保部医学科技青年培育计划孵化项目(20QNPY033)。

摘　　要：目的:探讨补体C5a/C5aR通路在小鼠缺血/再灌注肾损伤(I/R-AKI)中的作用及机制。方法:40只野生型小鼠随机分为假手术组、缺血20 min组、缺血30 min组、缺血40 min组;野生型和C5aR基因敲除小鼠各20只分为假手术组和缺血40 min组,再灌注24 h后取血清及肾组织。全自动生化分析仪检测BUN和Cre水平;RT-PCR检测肾组织C5aR、KIM-1、CXCL2、MIP-1α和CXCR2表达;ELISA检测血清C5a水平;HE明确肾损伤程度;IHC检测C5aR、KIM-1、Ccap3和Cyt C表达;IF检测中性粒细胞浸润;HK-2细胞体外模拟缺氧复氧环境,IF及Western blot检测C5aR的表达。结果:小鼠I/R-AKI模型中,随着缺血时间延长,BUN、Cre及血清C5a逐渐升高,肾脏病理损伤加重,肾组织C5aR表达逐渐增强,且C5a及C5aR表达与肾损伤程度正相关;体外缺氧复氧模型中,随着缺氧时间延长C5aR表达逐渐升高;与野生型小鼠相比,C5aR基因敲除小鼠BUN和Cre均明显减少,肾小管损伤减轻,KIM-1、Ccap3和Cyt C的表达下降;此外,C5aR基因敲除后CXCL2、MIP-1α和CXCR2表达降低,中性粒细胞浸润减少。结论:补体C5a/C5aR通路介导I/R-AKI病理过程,可能与其促进中性粒细胞浸润有关。Objective:To investigate the effects and underlying mechanisms of complement C5a/C5aR pathway on ischemiareperfusion induced acute kidney injury(I/R-AKI)in mice.Methods:Forty wild-type mice were divided into sham,20 min,30 min and 40 min ischemic groups;twenty wild-type and twenty C5aR knockout mice were divided into sham and 40 min ischemic groups,then the blood and kidney tissues were collected at 24 h after reperfusion. Automatic biochemical analyzer was used to measure BUN and Cre levels. C5aR,KIM-1,CXCL2,MIP-1α and CXCR2 expressions were tested by RT-PCR. C5a levels.were measured byELISA.Kidney pathological injury was measured by HE staining. C5aR,KIM-1,Ccap3 and Cyt C expressions were measured by IHC staining.The infiltration of neutrophils was observed by IF. Hypoxic re-oxygenation treatment to HK-2 cells in vitro was performed. The expression of C5aR in HK-2 cells was detected by IF and Western blot.Results:BUN,Cre,serum C5a levels,renal injury and C5aR expressions were significantly increased depending on the duration of ischemic time in I/R-AKI model. Moreover,C5a and C5aR expressions were positively correlated with the renal function and tubular injury. Consistently,the expression of C5aR increased gradually with prolonged hypoxia time in HK-2 cells in vitro. C5aR deficiency significantly attenuated renal ischemia reperfusion injury,as evidenced by decreased BUN and Cre,alleviated tubular damage scores,down-regulated KIM-1,Ccap3 and Cyt C expression,impaired CXCL2,MIP-1α and CXCR2 expression and attenuated neutrophils infiltration,compared with that of in wild-type mice.Conclusion:C5a/C5aR pathway mediates I/R-AKI,which mary be related to its effect on promoting and neutrophils infiltration.

关 键 词：缺血再灌注损伤 补体C5a/C5aR 中性粒细胞 

分 类 号：R692.9[医药卫生—泌尿科学]