补体C3a受体在db/db小鼠糖尿病肾病发病中的作用  

Role of complement C3a receptor in the pathogenesis of diabetic nephropathy in db/db mice

作　　者：林恩琴 张小红[2,3,4] 翁梦洁 郑晶 万建新 Lin Enqin;Zhang Xiaohong;Weng Mengjie;Zhen Jing;Wan Jianxin(The First Clinical College of Fujian Medical University,Fuzhou 350005,China;Department of Nephrology,Blood Purification Research Center,the First Affiliated Hospital,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;National Regional Medical Center,Binhai Campus of the First Affiliated Hospital,Fujian Medical University,Fuzhou 350212,China)

机构地区：[1]福建医科大学第一临床医学院,福州350005 [2]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 [3]福建省代谢性慢性肾病临床医学研究中心,福州350005 [4]福建医科大学附属第一医院滨海院区国家区域医疗中心,福州350212

出　　处：《中华肾脏病杂志》2024年第6期465-474,共10页Chinese Journal of Nephrology

基　　金：福建省卫健委青年科研项目(2020QNB025);福建省自然科学基金(2022J01212)。

摘　　要：目的探讨补体C3a受体在db/db小鼠糖尿病肾病发病中的作用,为糖尿病肾病的防治提供新靶点。方法12只8周龄雄性2型糖尿病(db/db)小鼠及6只同窝野生型(db/m)小鼠饲养于无特定病原体级环境。实验分组及干预:db/m组、db/db组、C3a受体拮抗剂组,每组6只,其中C3a受体拮抗剂组为db/db小鼠腹腔注射C3a受体拮抗剂(SB290157,10 mg/kg)进行干预,2 d腹腔注射1次,连续注射8周。收集血、尿样本,检测小鼠体重、血糖、血肌酐、血尿素氮、尿微量白蛋白/尿肌酐、尿N-乙酰-β-D-氨基葡萄糖苷酶(NAG)水平;收集肾组织,采用HE、PAS、Masson染色观察肾组织病理变化,免疫组化、免疫荧光及Western印迹检测肾组织C3及C3a受体表达,Western印迹检测肾组织肾损伤分1(Kim-1)、α平滑肌肌动蛋白(α-SMA)、紧密连接蛋白1(ZO-1)、波形蛋白、E钙黏蛋白(E-cadherin)的表达,免疫荧光分析α-SMA、ZO-1、Kim-1表达及分布,免疫组化分析白细胞介素1(IL-1)、肿瘤坏死因子α(TNF-α)表达,TUNEL法检测肾组织细胞凋亡情况。结果与db/m组相比,db/db组小鼠体重、空腹血糖、尿微量白蛋白/尿肌酐及尿NAG均明显较高(均P<0.01);与db/db组相比,C3a受体拮抗剂组小鼠体重、空腹血糖、尿微量白蛋白/尿肌酐及尿NAG均较低(均P<0.01);三组血肌酐和血尿素氮的差异均无统计学意义(均P>0.01)。与db/m组相比,db/db组小鼠肾小球肥大,肾小管上皮细胞坏死、脱落,肾小管扩张,C3、C3a受体蛋白表达均明显较高(均P<0.01);与db/db组相比,C3a受体拮抗剂组肾小球病变轻微,肾小管上皮细胞轻度坏死,肾小管扩张不明显。db/db组小鼠肾组织Kim-1、IL-1、TNF-α蛋白表达均高于db/m组,而C3a受体拮抗剂组Kim-1、IL-1、TNF-α蛋白表达均低于db/db组(均P<0.01)。与db/m组相比,db/db组小鼠肾小管上皮细胞α-SMA、波形蛋白蛋白表达均明显较高,ZO-1、E-cadherin蛋白表达均明显较低(均P<0.01);与db/db组相�Objective To investigate the role of complement C3a receptor in the diabetic nephropathy pathogenesis of db/db mice,and to provide a new target for prevention and treatment of diabetic nephropathy.Methods Twelve 8-week-old male mice with type 2 diabetes mellitus(db/db mice)and 6 wild-type(db/m)mice were reared in the special pathogen free environment.The mice were grouped into db/m group,db/db group and C3a receptor antagonist group,with 6 mice in each group.db/db model mice were intraperitoneally injected with C3a receptor antagonist(SB290157,10 mg/kg)once every two days for 8 weeks in C3a receptor antagonist group.Blood and urine samples were collected,and body weight of mice,fasting blood glucose,serum creatinine,blood urea nitrogen,urinary microalbumin/urinary creatinine and urinary N-acetyl-β-D-glucosaminidase(NAG)were detected.Renal tissues were collected,and HE,PAS and Masson stainings were used to observe the pathological changes.Immunohistochemistry,immunofluorescence and Western blotting were used to detect the protein expression levels of C3 and C3a receptor.Western blotting was used to analyze the protein expression levels of kidney injury molecule-1(Kim‐1),α-smooth muscle actin(α‐SMA),zonula occluden-1(ZO‐1),vimentin and E‐cadherin in renal tissues.Immunofluorescence was used to analyze the protein expression levels and distribution ofα‐SMA,ZO‐1 and Kim‐1,and immunohistochemistry was used to analyze the protein expression levels of interleukin-1(IL‐1)and tumor necrosis factor-α(TNF‐α).TUNEL assay was used to detect apoptotic cells in renal tissues.Results Compared with db/m group,body weight,fasting blood glucose,urinary microalbumin/urinary creatinine and urinary NAG in db/db group were significantly higher,while these indicators in C3a receptor antagonist group were slightly lower than those in db/db group(all P<0.01).There were no significant differences in serum creatinine and blood urea nitrogen among the three groups(all P>0.01).Compared with db/m group,db/db group had g

关 键 词：补体C3a 糖尿病肾病 细胞凋亡 炎症 

分 类 号：R692.9[医药卫生—泌尿科学] R587.2[医药卫生—外科学]