摘要
目的研究加味芪黄饮减轻糖尿病肾病(diabetic kidney disease,DKD)线粒体损伤及胰岛素抵抗的沉默信息调节因子2相关酶1(SIRT1)/上游信号抑癌基因(p53)/分子发动相关蛋白1(Drp1)轴的影响及机制。方法40只SPF级C57BL/6小鼠,其中30只小鼠用于建立糖尿病肾病模型设为DKD组,10只不建立糖尿病肾病模型设作为对照组;建模成功后再将DKD组中小鼠随机分为模型组、低剂量组、高剂量组,每组各10只;低剂量组、高剂量组分别给予加味芪黄饮400、800 mg/(kg·d),对照组、模型组小鼠给予等体积生理盐水,均为灌胃给药12周后处死,留取小鼠血、尿标本测定尿白蛋白肌酐比值(UACR);尿素氮(BUN)、血肌酐(Scr)、β2微球蛋白(β2-MG)、胱抑素C(Cys-C);血糖仪检测空腹血糖(FBG),放射免疫法检测空腹胰岛素(FIns)水平,计算胰岛素抵抗指数(HOMA-IR)等指标,采用DCFH-DA探针检测肾脏细胞中活性氧(ROS)水平、MitoSOXtmRed探针检测肾脏细胞线粒体中超氧化物歧化酶(SOD)、荧光检测ATP,HE染色观察大鼠肾组织形态,蛋白免疫印迹(Western Blot)检测肾组织中SIRT1、p53、Drp1水平。结果与对照组小鼠比较,模型组小鼠UACR、BUN、Scr、β2-MG、Cys-C、FBG、FIns、HOMA-IR、ROS、p53、Drp1水平升高、SOD、ATP、SIRT1下降(P<0.05);与模型组比较,低剂量组、高剂量组小鼠UACR、BUN、SCr、β2-MG、Cys-C、FBG、FIns、HOMA-IR、ROS、p53、Drp1水平下降、SIRT1、SOD、ATP升高(P<0.05),且高剂量组小鼠UACR、BUN、SCr、β2-MG、Cys-C、FBG、FIns、HOMA-IR、ROS、p53、Drp1水平低于低剂量组,SOD、ATP、SIRT1高于低剂量组(P<0.05)。结论加味芪黄饮可延缓糖尿病肾病进展,其机制可能与介导SIRT1/p53/Drp1轴减轻肾脏线粒体损伤,改善胰岛素抵抗状态有关。
Objective To study the effect and mechanism of Jiawei Qihuang Decoction(加味芪黄饮)reducing mitochondrial damage and insulin resistance silent information regulator 2 related enzyme 1(SIRT1)/upstream signal tumor suppressor gene(p53)/dynamin related protein 1(Drp1)axis of diabetic kidney disease(DKD).Methods Forty SPF C57 BL/6 mice were chosen,30 of which were used to establish the DKD model and used as the DKD group,and 10 of which without DKD model were used as the control group.After successful modeling,the mice in DKD group were randomly divided into model group,low-dose group and high-dose group with 10 mice in each group.Through intragastric administration,low-dose group and high-dose group were given 400 mg/(kg·d)and 800 mg/(kg·d)of Jiawei Qihuang Decoction respectively,and mice in the control group and model group were given the equal volume of normal saline.All mice were sacrificed after 12 weeks.Blood and urine samples of mice were collected to determine the levels of urine albumin-creatinine ratio(UACR),urea nitrogen(BUN),blood creatinine(Scr),β2 microglobulin(β2-MG),Cystatin C(Cys-C)and fasting blood glucose(FBG)were detected by blood glucose meter.The fasting insulin(FIns)level was detected by radioimmunoassay.The homeostasis model assessment-insulin resistance index(HOMA-IR)and other indicators were calculated.DCFH-DA probe was used to detect the level of reactive oxygen species(ROS)in kidney cells and MitoSOXtmRed probe to detect the superoxide dismutase(SOD)in kidney cell mitochondria.The fluorescence was used to detect the ATP.HE staining was used to observe the morphology of rat kidney tissue and Western Blot to detect the levels of SIRT1,p53 and Drp1 in kidney tissue.Results Compared with those of the control mice,the levels of UACR,BUN,Scr,β2-MG,Cys-C,FBG,FIns,HOMA-IR,ROS,p53 and Drp1 were increased in the model group mice,while the levels of SOD,ATP and SIRT1 decreased(P<0.05).Compared with those of the model group,the levels of UACR,BUN,SCr,β2-MG,Cys-C,FBG,FIns,HOMA-IR,ROS,p53 and Drp1 in the low-dose group and high-dose group were decreased,whereas the levels of SIRT1,SOD and ATP increased(P<0.05),and the levels of UACR,BUN,SCr,β2-MG,Cys-C,FBG,FIns,HOMA-IR,ROS,p53 and Drp1 were lower in the high-dose mice than those in the low-dose group,but the levels of SOD,ATP and SIRT1 were higher than those in the low-dose group(P<0.05).Conclusion Jiawei Qihuang Decoction can delay the progression of DKD,and its mechanism may be related to mediating the SIRT1/p53/Drp1 axis to reduce kidney mitochondrial damage and improve insulin resistance.
作者
蒙向欣
李辉远
李娟
周艳利
MENG Xiangxin;LI Huiyuan;LI Juan;ZHOU Yanli(Affiliated Hospital of Traditional Chinese Medicine of Guangzhou Medical University,Guangzhou 510130,Guangdong,China)
出处
《中华中医药学刊》
CAS
北大核心
2022年第7期193-197,I0032,I0033,共7页
Chinese Archives of Traditional Chinese Medicine
基金
广东省中医药管理局项目(20171225)。
关键词
加味芪黄饮
糖尿病肾病
线粒体损伤
胰岛素抵抗
足细胞沉默信息调节因子2相关酶1
上游信号抑癌基因
分子发动相关蛋白1
Jiawei Qihuang Decoction(加味芪黄饮)
diabetic kidney disease
mitochondrial damage
insulin resistance
podocyte silent information regulator 2 related enzyme 1
upstream signal tumor suppressor gene
molecular launch related protein 1
