大黄糖络丸调控AGEs/RAGE/IKK/NF-κB通路改善糖尿病肾病小鼠肾脏炎症损伤的机制  

Mechanism of Dahuang Tangluo Pills in Improving Renal Inflammatory Injury in Diabetic Kidkdey Disease by Regulating AGEs/RAGE/IKK/NF-κB Pathway

作　　者：章溥 梁建庆[1] 杨霞 白敏 朱向东 薛春霞 苏蓓蓓 赵芸慧 ZHANG Pu;LIANG Jianqing;YANG Xia;BAI Min;ZHU Xiangdong;XUE Chunxia;SU Beibei;ZHAO Yunhui(Gansu University of Chinese Medicine,Lanzhou 730000,China;Key Laboratory of Traditional Chinese Medicine(TCM)Prevention and Treatment of Regional High-incidence Diseases of the Ministry of Education in Ningxia,Yinchuan 750004,China;Tianshui TCM Hospital,Tianshui 741000,China)

机构地区：[1]甘肃中医药大学,兰州730000 [2]宁夏区域高发病中医药防治教育部重点实验室,银川750004 [3]天水市中医院,甘肃天水741000

出　　处：《中国实验方剂学杂志》2024年第20期77-85,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：甘肃省教育厅产业支撑计划项目(2021CYZC-03);2022年度甘肃中医药大学“中医老年病学”学科建设项目(2022-3)。

摘　　要：目的:探讨大黄糖络丸对db/db小鼠早期糖尿病肾病(DKD)的保护作用。方法:取8只db/m小鼠作为空白组,取40只雄性db/db小鼠通过尾静脉取血,检测空腹血糖(FBG),FBG≥16.7 mmol·L^(-1),尿量增多,且持续出现白蛋白尿提示造模成功。造模成功后随机分为模型组、达格列净组(1.5 mg·kg^(-1)·d^(-1))、大黄糖络丸高、中、低剂量组(3.6、1.8、0.9 g·kg^(-1)·d^(-1)),每组8只。各给药组均采用灌胃给药,空白组和模型组每日给予等体积蒸馏水灌胃,连续给药10周,观察小鼠生存状态并测定小鼠体质量、FBG及肾功能相关指标;酶联免疫吸附测定法(ELISA)检测肾组织中炎症相关指标;苏木素-伊红(HE)、马松(Masson)染色、电镜观察各组肾脏组织病理学改变;免疫荧光观察晚期糖基化终末产物(AGEs)、晚期糖基化终末产物受体(RAGE)蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测各组小鼠肾组织中AGEs、RAGE、核转录因子-κB(NF-κB)抑制蛋白激酶(IKK)、NF-κB mRNA和蛋白表达水平。结果:与空白组比较,模型组小鼠体质量、FBG、血清肌酐(SCr)、尿微量白蛋白/尿肌酐(ACR)、总胆固醇(TC)、甘油三酯(TG)均明显增高(P<0.05);肾组织中细胞间黏附分子-1(ICAM-1)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)明显增加(P<0.05);肾脏病理染色、电镜显示肾组织排列疏松,出现间隙,结构紊乱,系膜增生,纤维化明显;Real-time PCR结果显示,肾组织中RAGE、IKK、NF-κB mRNA表达明显增加(P<0.05);免疫荧光结果显示,肾组织中AGEs、RAGE蛋白表达明显增加(P<0.05);Western blot结果显示,肾组织中AGEs、RAGE、IKK、NF-κB蛋白表达明显增加(P<0.05)。给药干预后,与模型组比较,达格列净组和大黄糖络丸高剂量组小鼠体质量、FBG、SCr、ACR明显降低(P<0.05);达格列净组和大黄糖络丸高剂量组小鼠血清中TC明显降低(P<0.05);大黄糖络丸高剂�Objective:To explore the protective effects of Dahuang Tangluo pills on early diabetic kidkdey disease(DKD)in db/db mice.Method:Eight db/m mice were selected as the control group.Forty male db/db mice were selected and blood samples were collected via tail vein to measure fasting blood glucose(FBG).Mice with FBG≥16.7 mmol·L^(-1),increased urine output,and persistent albuminuria were considered successful in model establishment.After successful modeling,they were randomly divided into a model group,a dapagliflozin group(1.5 mg·kg^(-1)·d^(-1)),and high,medium,and low dose groups of Dahuang Tangluo pills(3.6,1.8,0.9 g·kg^(-1)·d^(-1),respectively),with eight mice in each group.All medication groups were administered orally,while the control and model groups were given an equal amount of distilled water by gavage daily.After continuous administration for 10 weeks,the survival status of the mice was observed,and their body weight,FBG,and kidney function-related indicators were measured.Inflammatory indicators in renal tissues were determined by enzyme-linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)staining,Masson staining,and electron microscopy were used to observe the pathological changes in renal tissues in each group.Immunofluorescence was employed to examine the expression of advanced glycation end products(AGEs)and receptors for advanced glycation end products(RAGE)proteins.Real-time quantitative polymerase chain reaction(Real-time PCR)and Western blot were utilized to detect the gene and protein expression levels of AGEs,RAGE,inhibitor of nuclear factor-κB(NF-κB)kinase(IKK),and NF-κB in the renal tissues of mice in each group.Result:Compared with control group,the model group showed a significant increase in body weight,FBG,serum creatinine(SCr),urinary microalbumin/urine creatinine ratio(ACR),total cholesterol(TC),and triglycerides(TG)(P<0.05).The levels of intercellular adhesion molecule-1(ICAM-1),interleukin-6(IL-6),and tumor necrosis factor-alpha(TNF-α)in renal tissues were significantly e

关 键 词：大黄糖络丸 糖尿病肾病 2型糖尿病 晚期糖基化终末产物(AGEs)/晚期糖基化终末产物受体(RAGE)/核转录因子-κB(NF-κB)抑制蛋白激酶(IKK)/NF-κB 炎症 

分 类 号：R2-0[医药卫生—中医学] R33R289R692