膜肾1号方对大鼠肾脏病理及PI3K/Akt/mTOR信号通路表达的影响  

The effect of membranous kidney No.1 formula on renal pathology and PI3K/Akt/mTOR signaling pathway expression in rats

作　　者：孟曦[1] 丁伟 王建美[3] 王耀光 MENG Xi;DING Wei;WANG Jianmei;WANG Yaoguang(Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital,Tianjin 300120,China;Tianjin Hedong Traditional Chinese Medicine Hospital,Tianjin 300162,China;Tianjin Beichen Hospital,Tianjin 300400,China;The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China)

机构地区：[1]天津市中医药研究院附属医院,天津300120 [2]天津市河东中医院,天津300162 [3]天津市北辰医院,天津300400 [4]天津市中医药大学第一附属医院,天津300193

出　　处：《天津中医药》2023年第10期1320-1326,共7页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81573888);国家重点研发计划项目(2018YFC1704102);中医药事业传承与发展:王耀光天津市名中医传承工作室项目(883022);天津市卫生健康委员会中医中西医结合科研课题项目(2021038)。

摘　　要：[目的]观察膜肾1号方对膜性肾病大鼠肾脏病理的改善作用及其对自噬通路磷脂肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/雷帕霉素靶蛋白(mTOR)相关蛋白表达的影响。[方法]将大鼠随机分为对照组、模型组、膜肾1号方高剂量组、中剂量组、低剂量组,盐酸贝那普利组。采用大鼠尾静脉注射阳离子化牛血清白蛋白(C-BSA)的方法建立MN大鼠模型,灌胃、取材。苏木精-伊红(HE)染色法观察大鼠肾脏组织病理改变;免疫球蛋白G(IgG)免疫荧光染色观察大鼠IgG沉积;蛋白免疫印迹法(Western Blot)检测PI3K/Akt/mTOR信号通路相关蛋白及自噬相关蛋白轻链3(LC3)表达。[结果]药物干预后,膜肾1号方组大鼠24 h尿蛋白、三酰甘油、总胆固醇、低密度脂蛋白下降且低于模型组,并具有统计学差异(P<0.05)。光镜下观察,HE染色示正常组肾组织整体结构基本正常,膜性肾病(MN)模型组肾小球毛细血管丛充血,系膜增生,基底膜出现增厚,部分肾小管细胞空泡变、组织内可见炎症细胞浸润,可见嗜复红蛋白及IgG沉积。经膜肾1号方和盐酸贝那普利干预后,大鼠肾脏病理学改变均有所减轻。各组大鼠IgG沉积显示,与对照组比较模型组IgG沉积明显,IgG荧光表达升高,差异具有统计学意义(P<0.05),盐酸贝那普利组和膜肾1号方组IgG荧光表达下降且低于模型组,具有统计学差异(P<0.05)。Western Blot检测显示,药物干预后,盐酸贝那普利组和膜肾1号方组大鼠PI3K-Akt信号通路相关蛋白表达下降,LC3表达增加,并具有统计学差异(P<0.05)。[结论]膜肾1号方可改善大鼠肾脏病理损伤,膜肾1号方干预后PI3K-Akt信号通路相关蛋白磷酸化磷酸肌醇3激酶(p-PI3K),磷酸化Akt蛋白(p-Akt),磷酸化雷帕霉素靶蛋白(p-mTOR)表达明显降低,自噬相关蛋白LC3表达升高,其分子机制与自噬信号通路的调控有关。[Objective]Observe the improvement effect of membranous kidney No.1 formula on renal pathology in rats with membranous nephropathy and its effect on the expression of autophagy pathway PI3K/Akt/mTOR related proteins.[Methods]Randomly divide the rats into a control group,a modeling group,a high-dose group of membranous kidney No.1 formula,a medium dose group,a low dose group,and a benazepril hydrochloride group.A membranous kidney rat model was established by injecting cationic bovine serum albumin(C-BSA)into the tail vein of rats,and the samples were taken by gavage.HE staining method was used to observe the pathological changes of rat kidney tissue;IgG immunofluorescence staining was used to observe the deposition of IgG in rats;Western blot was used to detect the expression of PI3K/Akt/mTOR signaling pathway related proteins and autophagy related protein LC3.[Results]After drug intervention,the 24-hour urinary protein,triglyceride,total cholesterol,high-density lipoprotein and low-density lipoprotein of rats in the membranous kidney No.1 formula group decreased and were lower than those in the model group,with statistical differences(P<0.05).Under the light microscope,HE staining showed that the overall structure of the normal group’s renal tissue was basically normal.In the membranous kidney model group,the glomerular capillary plexus was congested,the mesangial hyperplasia was observed,the basement membrane was thickened,some renal tubular cells were vacuolated,inflammatory cell infiltration was observed in the tissue,and eosinophil and IgG deposition were observed.After intervention with membranous kidney No.1 formula and benazepril hydrochloride,the pathological changes in the kidneys of rats were alleviated.The IgG deposition of rats in each group showed that compared with the normal group,the model group had significant IgG deposition and significantly increased IgG fluorescence expression(P<0.05).The IgG fluorescence expression of the benazepril hydrochloride group and the membranous kidney No.1 formul

关 键 词：膜肾1号方 PI3K/Akt/mTOR信号通路 膜性肾病 

分 类 号：R692[医药卫生—泌尿科学]