基于Nrf2/HO-1通路探讨金雀异黄酮对糖尿病肾病大鼠肾损伤的影响及机制  

Effect and mechanism of genistein on renal injury in diabetic nephropathy rats via Nrf2/HO-1 pathway

作　　者：陈洁[1] 杨林燕 郝晓娟[1] 何琦 侯世杰 CHEN Jie;YANG Linyan;HAO Xiaojuan;HE Qi;HOU Shijie(Handan Central Hospital,Handan 056001,Hebei,China)

机构地区：[1]邯郸市中心医院,河北邯郸056001

出　　处：《现代中西医结合杂志》2023年第16期2241-2248,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家“十三五”科技支撑计划项目(2016YFC1319026);河北省医学科学研究课题(20211577)。

摘　　要：目的探讨金雀异黄酮对糖尿病肾病大鼠肾组织损伤的影响及其可能的分子机制。方法取雄性SD大鼠55只,随机选择10只作为正常组,其余大鼠采用一次性腹腔注射链脲佐菌素60 mg/kg的方法复制糖尿病肾病模型。将40只成模大鼠随机分为模型组、金雀异黄酮组、鸦胆子苦醇组和金雀异黄酮+鸦胆子苦醇组,每组10只。金雀异黄酮组给予30 mg/kg金雀异黄酮灌胃,鸦胆子苦醇组给予鸦胆子苦醇2 mg/kg灌胃,金雀异黄酮+鸦胆子苦醇组给予30 mg/kg金雀异黄酮和鸦胆子苦醇2 mg/kg灌胃,均1次/d,连续灌胃6周。检测各组大鼠空腹血糖(FPG)水平及肾功能指标[血清肌酐(SCr)、尿素氮(BUN)和24 h尿微量白蛋白(24h U-mAlb)],计算肾脏指数,HE染色、PAS染色、Masson染色观察肾组织形态,透射电子显微镜观察肾细胞超微结构,分光光度法检测肾组织中总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量,ELISA法检测肾组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)含量,Western blot法检测肾组织中E2相关因子2(Nrf2)、p-Nrf2、血红素加氧酶1(HO-1)、醌氧化还原酶1(NQO1)、总核因子κB(NF-κB)、胞核NF-κB蛋白表达情况。结果与正常组比较,模型组大鼠FPG、SCr、BUN、24hU-mAlb水平和肾脏指数均明显升高(P均<0.05);肾小球肥大、硬化,炎性浸润,胶原沉积,肾小球细胞可见足突肿胀或消失、基底膜弥漫性增厚、线粒体嵴断裂或消失;肾组织中T-SOD、CAT活性均明显降低(P均<0.05),MDA、TNF-α、IL-1β、IL-8含量均明显升高(P均<0.05);肾组织中p-Nrf2、HO-1、NQO1蛋白相对表达量及p-Nrf2/Nrf2比值均明显降低(P均<0.05),胞核NF-κB蛋白相对表达量及胞核NF-κB/总NF-κB比值均明显升高(P均<0.05)。与模型组比较,金雀异黄酮组和金雀异黄酮+鸦胆子苦醇组大鼠FPG、SCr、BUN、24hU-mAlb水平和肾脏指数均明显降低(P均<0.05);肾组织Objective It is to explore the effects of genistein on renal tissue injury in rats with diabetic nephropathy and its possible molecular mechanisms.Methods Fifty-five male SD rats were taken,10 of which were randomly selected as the normal group,and the rest were used to replicate the diabetic nephropathy models by one-time intraperitoneal injection of streptozotocin 60 mg/kg.The 40 modeled rats were randomly divided into model group,genistein group,brusatol group and genistein+brusatol group,with 10 rats in each group.The rats in the genistein group were gavaged with 30 mg/kg of genistein,the rats in the brusatol group were gavaged with 2 mg/kg of brusatol,and the rats in the genistein+brusatol group were gavaged with 30 mg/kg of genistein and 2 mg/kg of genistein+brusatol,all of which were administered once daily,continuously treated for 6 weeks.The levels of fasting plasma glucose(FPG)and renal function indexes[serum creatinine(SCr),urea nitrogen(BUN)and 24 h urinary microalbumin(24hU-mAlb)]of rats in each group were detected,and their renal index was calculated,and the morphology of renal tissues was observed by HE staining,PAS staining,and Masson staining,and the ultrastructure of renal cells was observed by transmission electron microscope,the activities of total superoxide dismutase(T-SOD),catalase(CAT)and the contents of malondialdehyde(MDA)in renal tissues were detected by spectrophotometry,the contents of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-8(IL-8)in renal tissues were detected by ELISA,and the expressions of E2-related factor 2(Nrf2),p-Nrf2,heme oxygenase 1(HO-1),quinone oxidoreductase 1(NQO1),total nuclear factorκB(NF-κB),and cytosolic NF-κB proteins in renal tissues were detected by Western blot.Results Compared with the normal group,the levels of FPG,SCr,BUN,24hU-mAlb and renal index of rats in the model group were significantly increased(all P<0.05);the renal tissue presented pathological changes such as glomerular hypertrophy,matrix hyperplasia,mesangial expansio

关 键 词：金雀异黄酮 糖尿病肾病 肾损伤 血红素加氧酶1 E2相关因子2 氧化应激 炎症反应 

分 类 号：R-332[医药卫生]