文献类型：期刊文献

中文题名：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

英文题名：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[1];梁建庆[1];杨霞[3];白敏[1];朱向东[2];薛春霞[1];苏蓓蓓[1];赵芸慧[1]

第一作者：章溥

机构：[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

第一机构：甘肃中医药大学

年份：2024

卷号：30

期号：20

起止页码：77

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

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)。

语种：中文

中文关键词：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

外文关键词：Dahuang Tangluo pills;diabetic nephropathy;type 2 diabetes mellitus;advanced glycation end products(AGEs)/receptors for advanced glycation end products(RAGE)/inhibitor of nuclear factor-κB(NF-κB)kinase(IKK)/NF-κB;inflammation

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);大黄糖络丸高剂量组小鼠血清中TG明显降低(P<0.05);各给药组肾组织中ICAM-1、IL-6和TNF-α表达明显降低(P<0.05)。肾脏病理染色、电镜显示各给药组小鼠肾脏损伤减轻,胶原纤维沉积减轻,系膜增生减少;Real-time PCR结果显示达格列净组和大黄糖络丸高、中剂量组肾组织中RAGE、IKK、NF-κB mRNA表达明显降低(P<0.05);免疫荧光结果显示,达格列净组和大黄糖络丸高、中剂量组肾组织中AGEs、RAGE蛋白表达明显降低(P<0.05);Western blot结果显示,达格列净组和大黄糖络丸高、中剂量组肾组织中AGEs、RAGE、IKK、NF-κB蛋白表达明显降低(P<0.05)。结论:大黄糖络丸能够改善DKD小鼠肾脏病理结构和肾脏炎症损伤,其作用机制可能与抑制AGEs/RAGE/IKK/NF-κB通路有关。
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 elevated(P<0.05).Renal histopathological staining and electron microscopy revealed loose arrangement,gaps,structural disarray,mesangial proliferation,and significant fibrosis in renal tissues.Real-time PCR results showed a significant increase in the expression of RAGE,IKK,and NF-κB genes in renal tissues(P<0.05).Immunofluorescence results demonstrated a significant increase in the expression of AGEs and RAGE proteins in renal tissues(P<0.05).Western blot results showed a significant increase in the expression of AGEs,RAGE,IKK,and NF-κB proteins in renal tissues(P<0.05).After drug intervention,compared with model group,the dapagliflozin group and the high-dose Dahuang Tangluo pills group showed significant reductions in body weight,FBG,SCr,and ACR(P<0.05),and a significant decrease in TC in mouse serum(P<0.05),while the high-dose Dahuang Tangluo pills group showed a significant decrease in TG in mouse serum(P<0.05).All treatment groups showed a significant reduction in ICAM-1,IL-6,and TNF-αin renal tissues(P<0.05).Renal histopathological staining and electron microscopy showed improved kidney injury,decreased collagen fiber deposition,and reduced mesangial proliferation in all treatment groups.Real-time PCR results showed a significant decrease in the expression of RAGE,IKK,and NF-κB genes in the dapagliflozin group and the highand medium-dose Dahuang Tangluo pills groups(P<0.05).Immunofluorescence results demonstrated a significant decrease in the expression of AGEs and RAGE proteins in the dapagliflozin group and the high-and medium-dose Dahuang Tangluo pills groups(P<0.05).Western blot results showed a significant decrease in the expression of AGEs,RAGE,IKK,and NF-κB proteins in the dapagliflozin group and the high-and mediumdose Dahuang Tangluo pills groups(P<0.05).Conclusion:Dahuang Tangluo pills can improve the pathological structure of the kidneys and reduce renal inflammation in DKD mice,possibly through inhibiting the AGEs/RAGE/IKK/NF-κB pathway.