芪石肾舒胶囊对早期糖尿病肾病小鼠肾纤维化的 作用机制研究  

Effect and mechanism of Qishishenshu Capsule on renal fibrosis in mouse early diabetic nephropathy

作　　者：劳筱清 陈晨 张宏民 杨秀 史姜珊 粟宏伟 沈宏萍 王丽 游蔓芮 李晓斌 赵长英 LAO Xiaoqing;CHEN Chen;ZHANG Hongmin;YANG Xiu;SHI Jiangshan;SU Hongwei;SHEN Hongping;WANG Li;YOU Manrui;LI Xiaobin;ZHAO Changying(College of Integrated Traditional Chinese and Western Medicine,Southwest Medical University,Luzhou 646000,China;Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000;Leshan Hospital of Traditional Chinese Medicine,Leshan 614000)

机构地区：[1]西南医科大学中西医结合学院,四川泸州646000 [2]西南医科大学附属中医医院,四川泸州646000 [3]乐山市中医医院,四川乐山614000

出　　处：《中国比较医学杂志》2024年第9期56-65,共10页Chinese Journal of Comparative Medicine

基　　金：四川省科技计划资助(2022YFS0407,2022YFS0621);四川省中医药管理局(2021MS426)。

摘　　要：目的探讨芪石肾舒胶囊对早期糖尿病肾病小鼠肾纤维化的治疗作用和机制。方法采用多次注射链脲佐菌素建立糖尿病肾病小鼠模型。小鼠随机分为正常组、模型组和给药组(0.9 g/(kg·d)),每组8只,连续灌胃4周,监测每周空腹血糖。4周后,检测尿微量白蛋白肌酐比、血肌酐和血尿素氮含量;苏木精-伊红染色、过碘酸雪夫染色和天狼星红染色观察肾病理变化情况;实时荧光定量逆转录聚合酶链式反应检测纤维连接蛋白、Ⅰ型胶原蛋白α1及α-平滑肌肌动蛋白的mRNA表达水平;免疫组化和免疫蛋白印迹法检测纤维连接蛋白、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、α-平滑肌肌动蛋白、足突蛋白、肾病蛋白及转化生长因子-β1/SMAD家族成员2/3(TGF-β1/Smad2/3)通路相关蛋白含量。结果与正常组相比,模型组小鼠空腹血糖和尿微量白蛋白肌酐比水平升高(P<0.001);肾组织出现系膜增生、基底膜增厚和胶原沉积;纤维连接蛋白、Ⅰ型胶原蛋白α1及α-平滑肌肌动蛋白的mRNA水平升高(P<0.05);足突蛋白和肾病蛋白水平下降(P<0.05),纤维连接蛋白、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、α-平滑肌肌动蛋白、足突蛋白、肾病蛋白及TGF-β1/Smad2/3通路蛋白水平升高(P<0.05);与模型组相比,给药组尿微量白蛋白肌酐比水平降低(P<0.05),肾病理损伤有所缓解;纤维连接蛋白、Ⅰ型胶原蛋白α1及α-平滑肌肌动蛋白的mRNA水平下降(P<0.05),足突蛋白和肾病蛋白水平升高(P<0.05),纤维连接蛋白、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、α-平滑肌肌动蛋白及TGF-β1/Smad2/3通路蛋白水平均降低(P<0.05)。结论芪石肾舒胶囊可改善糖尿病肾病小鼠肾纤维化,其机制可能与抑制TGF-β1/Smad2/3信号通路有关。Objective To investigate the therapeutic effect and underlying mechanism of Qishishenshu Capsule on renal fibrosis in mice with early diabetic nephropathy(DN).Methods A DN mouse model was established by multiple injections of streptozotocin.The mice were randomly divided into a normal group(NC),model group(DN),and Qishi group(QS)(0.9 g/(kg·d)),with eight mice in each group.Mice were gavaged continuously for 4 weeks,and fasting blood glucose(FBG)was measured weekly.Four weeks later,urinary albumin/creatinine(UACR),serum creatinine,and blood urea nitrogen were measured.Hematoxylin-eosin,periodicacid-Schiff,and Sirius red staining were used to analyze renal pathological changes.Real-time fluorescence quantitative reverse-transcription polymerase chain reaction was used to detect the mRNA levels of fibronectin(FN),collagen type Ⅰ alpha 1(Col1a1),andα-smooth muscle actin(α-SMA).Immunohistochemistry and Western blot were performed to detect FN,collagen type Ⅰ(Collagen Ⅰ),collagen type Ⅲ(Collagen Ⅲ),α-SMA,Podocin,Nephrin,and transforming growth factor-β1/SMAD family member2/3(TGF-β1/Smad2/3)pathway-related proteins.Results Compared with mice in the NC group,those in the DN group showed significantly higher levels of FBG and UACR(P<0.001),and mesangial hyperplasia,basement membrane thickening,and collagen deposition in the renal tissue.The mRNA levels of FN,Col1a1,and α-SMA were increased(P<0.05).Protein levels of Podocin and Nephrin were decreased(P<0.05).The levels of FN,Collagen Ⅰ,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were increased(P<0.05).Compared with the DN group,the QS group’s level of UACR was decreased(P<0.05),their renal pathological injury was alleviated,and mRNA levels of FN,Collagen Ⅰ,and α-SMA were attenuated(P<0.05);whereas their protein levels of Podocin and Nephrin were elevated(P<0.05).The levels of FN,Collagen Ⅰ,Collagen III,α-SMA,and TGF-β1/Smad2/3 pathway proteins were also decreased(P<0.05).Conclusions Qishishenshu Capsule improved renal fibrosis in DN

关 键 词：芪石肾舒胶囊 糖尿病肾病 肾纤维化 足细胞 TGF-β1/Smad2/3 

分 类 号：R-33[医药卫生]