藏族药灰兜巴下调Nox4表达减轻糖尿病肾病大鼠肾脏氧化应激损伤  

Huidouba Ameliorates Kidney Oxidative Stress Injury by Down-regulating Nox4 Expression in Rats with Diabetic Nephropathy

作　　者：杨坤宝 庞宗然[1] 白颖慧 鲁碧楠 于宁 韩桂艳[2] 尹长江 YANG Kun-bao;PANG Zong-ran;BAI Ying-hui;LU Bi-nan;YU Ning;HAN Gui-yan;YIN Chang-jiang(College of Pharmacy,Minzu University of China,Beijing 100081,China;Chengde Medical University,Chengde 067000,China)

机构地区：[1]中央民族大学药学院,北京100081 [2]承德医学院,河北承德067000

出　　处：《中国实验方剂学杂志》2020年第18期84-90,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81804217);承德医学院重点课题项目(201810);承德医学院重大项目科研专项(KY2020006)。

摘　　要：目的:从减轻糖尿病肾病(DN)大鼠肾脏氧化应激损伤角度探讨藏族药灰兜巴的肾脏保护作用,为其治疗DN提供科学依据。方法:本实验采取高脂饮食喂养加腹腔注射链脲佐菌素(STZ)65 mg·kg-1法诱导DN大鼠模型,空白组注射等体积溶媒并饲以大鼠维持饲料。将造模成功DN大鼠随机分为DN模型组(10 mL·kg-1·d-1),二甲双胍组(0.045 g·kg-1·d-1),灰兜巴组(0.18 g·kg-1·d-1),每组8只,分别以溶媒或相应药物灌胃给药干预6周。动态监测各组大鼠体质量、血糖变化,并于药物干预前后收集24 h尿液,检测24 h尿微量白蛋白(mAlb);实验取材后,计算肾脏质量指数(KI);并利用全自动生化分析仪检测大鼠血肌酐(SCr)和血尿素氮(BUN)水平;马松(Masson)染色观察肾组织病理学改变;脂质氧化丙二醛(MDA)测定试剂盒检测肾组织MDA含量;蛋白免疫印迹法(Western blot)和免疫荧光法检测肾组织烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶4(Nox4)蛋白表达。结果:与空白组比较,DN模型组大鼠空腹血糖(FBG)及24 h mAlb,SCr,BUN水平均显著升高(P<0.01),肾组织MDA含量显著增加(P<0.01),肾组织病理损伤明显,肾小球Nox4蛋白表达显著增多(P<0.01);与模型组比较,各给药组DN大鼠FBG及24 h mAlb,SCr,BUN水平显著下降(P<0.01),肾组织病理损伤及蛋白尿减轻,肾组织MDA含量显著降低(P<0.01),肾小球Nox4表达显著下调(P<0.01),肾脏氧化应激损伤得到改善。结论:益气养阴藏族药灰兜巴可减轻DN大鼠肾脏氧化应激损伤,降低蛋白尿,从而延缓DN发生发展,其作用机制与下调DN大鼠肾脏组织NADPH氧化酶Nox4蛋白表达有关。Objective:From a new perspective,to explore therapeutic effect of Huidouba(HDB)on alleviating kidney oxidative damage in rats with diabetic nephropathy(DN)and provide a scientific basis for developing HDB as a potential Tibetan medicine for treatment of DN.Method:Rats were fed with high-fat diet(HFD)and injected with streptozocin(STZ,65 mg·kg-1)intraperitoneally to induce DN model,while rats in Blank group were injected with an equal volume of vehicle and fed with normal chow.The successfully modeling DN rats were randomly divided into three groups,8 rats per group,DN model group(10 mL·kg-1·d-1),Metformin group(0.045 g·kg-1·d-1)and HDB group(0.18 g·kg-1·d-1).Monitor body weight(BW)and fasting blood glucose(FBG)weekly,and collect 24 hours urine before and after medication to examine microalbuminuria(mAlb).Calculate kidney index(KI)after sacrificing,analyze mAlb,serum creatinine(SCr)and blood urea nitrogen(BUN)with a fully automatic biochemical analyzer.Histopathology of kidney was observed by Masson staining.Lipid peroxidation malondialdehyde(MDA)assay kit was used to examine MDA content in kidney tissue.Nox4,as a subtype of triphosphopyridine nucleotide(NADPH)oxidase family was determined by Western blot and immunofluorescence assay of kidney tissue.Result:Compared with blank group,levels of FBG,24 h mAlb,SCr,BUN and MDA in DN model group were increased(P<0.01),tissue damage was obvious and Nox4 expression in glumeruli was increased significantly(P<0.01).Compared with DN model group,levels of FBG,24 h mAlb,SCr,BUN and MDA in drug administration groups were decreased(P<0.01),kidney injury was alleviated and Nox4 expression was down-regulated(P<0.01).Conclusion:HDB as a Yiqiyangyin Tibetan medicine,could ease oxidative stress injury of kidney and reduce proteinuria in DN rats,thus prevent the development of DN.Its mechanism is closely related to down-regulating Nox4 expression of kidney tissue in DN rats.

关 键 词：灰兜巴 烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶4(Nox4) 蛋白尿 糖尿病肾病 氧化应激 

分 类 号：R2-0[医药卫生—中医学] R587.1