补体C3在肾间质纤维化过程中参与募集、活化肥大细胞  

Complement C3 is involved in the recruitment and activation of mast cells during renal interstitial fibrosis

作　　者：冯艳萍 李镇洲[3,4,5] 崔炯 吴小婷[3,4,5] 杨丽燕 万建新 Feng Yanping;Li Zhenzhou;Cui Jiong;Wu Xiaoting;Yang Liyan;Wan Jianxin(Department of Nephrology,Fuzhou Second General Hospital,Fuzhou 350007,China;The First Clinical College of Fujian Medical University,Fuzhou 350005,China;Department of Nephrology,Blood Purification Research Center,Fujian Medical University,Fuzhou 350005,China;Fujian Clinical Research Center for Metabolic Chronic Kidney Disease,the First Affiliated Hospital,Fujian Medical University,Fuzhou 350005,China;Department of Nephrology,National Regional Medical Center,Binhai Campus of the First Affiliated Hospital,Fujian Medical University,Fuzhou 350212,China)

机构地区：[1]福州市第二总医院肾内科,福州350007 [2]福建医科大学第一临床医学院,福州350005 [3]福建医科大学附属第一医院肾内科血液净化研究中心,福州350005 [4]福建省代谢性慢性肾病临床医学研究中心,福州350005 [5]福建医科大学附属第一医院滨海院区国家区域医疗中心肾内科,福州350212

出　　处：《中华肾脏病杂志》2024年第12期952-960,共9页Chinese Journal of Nephrology

基　　金：福建省卫生健康青年科研课题(2021QNA023);福建省卫生健康医学创新课题(2022CXA025)。

摘　　要：目的观察补体C3在肾脏间质纤维化过程中的作用。方法8~12周龄的C57BL/6野生型(wild type,WT)和补体C3基因敲除(C3 gene knockout,C3KO)小鼠用随机数字表法分为:WT小鼠假手术组(WTcontrol)、WT小鼠单侧输尿管结扎(unilateral ureter obstruction,UUO)模型组(WTuuo)、C3KO小鼠假手术组(C3KOcontrol)和C3KO小鼠UUO模型组(C3KOuuo)。左侧输尿管结扎法建立UUO模型,每组6只。Masson及HE染色观察肾组织病理改变;免疫组化对肾组织C3、胰蛋白酶(tryptase)、血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)、转化生长因子β1(transforming growth factor⁃β1,TGF⁃β1)、基质金属蛋白酶9(matrix metalloproteinase⁃9,MMP⁃9)进行定位及半定量检测;免疫荧光法检测肾组织糜蛋白酶(chymase)水平;酶联免疫吸附测定法检测肾组织AngⅡ、C3裂解片段C3a、MMP⁃9的水平;实时定量PCR法检测肾组织肾素mRNA的变化;Western印迹法检测肾组织chymase、肾素、TGF⁃β1的变化。结果与WTcontrol组小鼠相比,WTuuo组小鼠肾组织肾小管出现明显损伤、肾间质纤维化,肥大细胞浸润增多,C3、C3a、chymase、肾素、AngⅡ、TGF⁃β1、MMP⁃9的表达均较高(均P<0.05);与WTuuo组小鼠相比,C3KOuuo组小鼠肾小管损伤、肾间质纤维化明显减轻,肾组织未检测到C3、C3a,肥大细胞浸润减少,chymase、肾素、AngⅡ、TGF⁃β1、MMP⁃9的表达均较低(均P<0.05)。结论C3/C3a在肾脏间质纤维化过程中可能参与募集、活化肥大细胞使其释放chymase并促进肾素、AngⅡ、TGF⁃β1、MMP⁃9等物质表达增加从而加重肾脏损伤。Objective To observe the role of complement C3 in the process of renal interstitial fibrosis.Methods Renal interstitial fibrosis model was established by unilateral ureteral obstruction(UUO)in male C3-deficient(C3KO)mice and age-matched C57BL/6 wild type(WT)mice(8-12 weeks of age).Mice were randomly divided into 4 groups,including sham operation in WT group(WTcontrol)(n=6),UUO operation in WT group(WTuuo)(n=6),sham operation in C3-deficient group(C3KOcontrol)(n=6),and UUO operation in C3-deficient group(C3KOuuo)(n=6).Tubular interstitial fibrosis was observed by both HE staining and Masson staining.The expression of C3,trypsin(tryptase),angiotensinⅡ(AngⅡ),transforming growth factorβ1(TGF-β1),and matrix metalloproteinase-9(MMP‐9)was detected by immunohistochemical staining.Chymase level were assessed by immunofluorescence staining.The levels of AngⅡand C3 cleavage fragments C3a and MMP‐9 were determined by enzyme-linked immunosorbent assay.The change in renin mRNA was determined by real-time PCR.The changes of chymase,renin,and TGF-β1 were detected by Western blotting.Results Compared with the WTcontrol group mice,the WTuuo group mice showed significant renal tubular injury,renal interstitial fibrosis,increased infiltration of mast cells,and significantly increased expression of C3,C3a,chymase,renin,AngⅡ,TGF-β1,and MMP‐9 in the renal tissue(all P<0.05).Compared with the WTuuo group mice,the renal tubular injury and renal interstitial fibrosis in the C3KOuuo group mice were significantly reduced,and C3 and C3a were not detected in renal tissue.Mast cells infiltration was reduced,and the expression of chymase,renin,AngⅡ,TGF-β1,and MMP‐9 was weakened(all P<0.05).Conclusion C3/C3a can participate in the recruitment and activation of mast cells to release chymase in kidney interstitial fibrosis,and promote the expression of renin,AngⅡ,TGF-β1,MMP 9 and other substances,thus aggravating kidney injury.

关 键 词：补体C3 纤维化 肥大细胞 肾间质 

分 类 号：R692[医药卫生—泌尿科学]