补体C3参与小鼠缺血再灌注急性肾损伤后肾脏炎症及慢性纤维化  

Complement C3 participates in inflammation and post-injury fibrosis in mice with ischemia-reperfusion induced acute kidney injury

作　　者：饶斯逸 张晶 崔炯[1] 万建新[1] 陈怡[1] RAO Si-yi;ZHANG Jing;CUI Jiong;WAN Jian-xin;CHEN Yi(Department of Nephrology,Blood Purification Research Center,The First Affiliated Hospital of Fujian Medical University,Fuzhou,Fujian 350005,China)

机构地区：[1]福建医科大学附属第一医院肾内科血液净化研究中心,福建福州350005

出　　处：《中华高血压杂志》2022年第2期145-152,共8页Chinese Journal of Hypertension

基　　金：福建省自然科学基金项目(2019J01682)。

摘　　要：目的探讨补体C3是否参与缺血再灌注急性肾损伤(AKI)后肾脏慢性纤维化过程。方法C57BL/6小鼠50只予微型动脉夹夹闭左侧肾动脉,持续30 min后恢复血流,1周后切除右侧肾脏,建立肾脏缺血再灌注损伤后慢性纤维化模型,随机分为5组,分别于第0、3、7、14、28天,酶联免疫吸附试验(ELISA)法和Western-blot法检测肾组织补体C3、白细胞介素(IL)1、IL-6、转化生长因子(TGF)-β1含量或表达水平。另取20只C57BL/6小鼠和20只周龄、性别与之相配对补体C3基因敲除小鼠(C3^(-/-)),各随机分为2组:肾脏缺血再灌注损伤后慢性纤维化组[C57BL/6-AKI-慢性肾脏病(CKD)组、C3^(-/-)-AKI-CKD组]及仅分离左肾动脉,不阻断的假手术(sham)组(C57BL/6-sham组、C3^(-/-)-sham组),28 d后检测24 h尿蛋白、血肌酐、尿素氮;苏木素-伊红(HE)染色和Masson染色观察肾小球、肾小管及肾间质纤维化;ELISA法测肾组织C3、IL-1、IL-6、TGF-β1含量;用Western-blot法测补体C3、IL-1、IL-6、TGF-β1蛋白表达水平。结果AKI后肾组织C3及炎症因子IL-1、IL-6、TGF-β1呈时间依赖性增加,第7天达到高峰(均P<0.01)。与C57BL/6-sham组、C3^(-/-)-sham组相比,第28天,C57BL/6-AKI-CKD组和C3^(-/-)-AKI-CKD组小鼠血肌酐[肌酐:C57BL/6-sham组(8.05±1.06)比C57BL/6-AKI-CKD组(25.35±3.02)比C3^(-/-)-sham组(7.96±1.09)比C3^(-/-)-AKI-CKD组(16.94±1.89)μmol/L,F=210.72,P<0.01]、尿素氮和尿蛋白升高,肾脏组织补体C3、炎症因子IL-1、IL-6、TGF-β1的表达增加,肾脏纤维化明显;与C57BL/6-AKI-CKD组相比,第28天,C3^(-/-)-AKI-CKD组小鼠血肌酐、尿素氮和尿蛋白水平降低,肾脏组织补体C3、炎症因子IL-1、IL-6、TGF-β1的表达水平降低,肾小球系膜增生、肾小球硬化和肾小管间质评分降低;肾小球、肾小管和肾间质胶原面积减少(均P<0.05)。结论补体C3参与了缺血再灌注AKI后肾脏炎症及慢性纤维化。Objective To investigate whether complement C3 is involved in the process of post-injury fibrosis after ischemia-reperfusion induced acute kidney injury(IR-AKI).Methods The IR-AKI and post-injury fibrosis model was established by left renal artery clamping with micro-arterial clip for 30 minutes,and resecting the right kidney one week later in C57 BL/6 mice.Fifty successful mice were randomly divided into 5 groups to detect the concentrations of complement C3,interleukin(IL)1,IL-6 and transforming growth factor(TGF)-β1 and the protein expression of these indexes in renal tissue by enzyme-linked immunosorbent assay(ELISA)and Western blot on day 0,3,7,14 and 28 respectively.In addition,twenty C57 BL/6 mice and twenty gender-and age-matched complement C3 knockout(C3^(-/-))mice were randomly divided into IR-AKI and post-injury fibrosis(chronic kidney disease,CKD)group and sham-operation group respectively,namely C57 BL/6-AKI-CKD group,C3^(-/-)-AKI-CKD group,C57 BL/6-sham group,C3^(-/-)-sham group.In the sham group,only the left renal artery were isolated without blocking the blood flow.After 28 days,the 24 h urine protein,serum creatinine(Scr)and blood urea nitrogen(BUN)were detected;HE and Masson staining were performed to observe glomerulus,renal tubules and renal fibrosis;the concentrations of complement C3,IL-1,IL-6 and TGF-β1 of renal tissue were measured by ELISA,and the expression of complement C3,IL-1,IL-6,and TGF-β1 protein were detected by Western blot.Results After AKI,the expression of renal complement C3,IL-1,IL-6 and TGF-β1 increased in a time-dependent manner and reached to the peak on the 7 th day.Compared with the sham group,on the 28 th day,the Scr[C57 BL/6-sham(8.05±1.06)vs C57 BL/6-AKI-CKD(25.35±3.02)vs C3^(-/-)-sham(7.96±1.09)vs C3^(-/-)-AKI-CKD(16.94±1.89)μmol/L,F=210.72,P<0.01],BUN and urine protein levels in C57 BL/6-AKI-CKD group and C3^(-/-)-AKI-CKD group increased,and the levels of renal complement C3,IL-1,IL-6 and TGF-β1 increased,and the renal fibrosis was obvious.Compared with

关 键 词：急性肾损伤 慢性肾脏病 补体C3 炎症反应 C3基因敲除 小鼠 

分 类 号：R692[医药卫生—泌尿科学]