OAT1/3在肾小管表达差异及竞争性抑制对急性马兜铃酸Ⅰ肾小管损伤的影响  

Effects of OAT1/3 differential expression and competitive inhibition on renal tubular injury induced by aristolochic acid I

作　　者：杨心怡 黄春华 李小芬 李茂娟 夏铭 汤海明 刘德芳 艾兴辉 楼迪栋 YANG Xinyi;HUANG Chunhua;LI Xiaofen;LI Maojuan;XIA Ming;TANG Haiming;LIU Defang;AI Xinghui;LOU Didong(Department of Forensic Medicine,Guizhou University of Chinese Medicine,Guiyang 550000,China;Judicial Appraisal Center,Guizhou University of Traditional Chinese Medicine,Guiyang 550000,China;Guizhou Provincial Key Laboratory of Forensic Medicine Toxicology,Guizhou University of Chinese Medicine Guiyang 550000,China)

机构地区：[1]贵州中医药大学基础医学院法医学教研室,贵州贵阳550000 [2]贵州中医药大学司法鉴定所,贵州贵阳550000 [3]贵州中医药大学贵州省法医中药毒理学特色重点实验室,贵州贵阳550000

出　　处：《药物评价研究》2024年第10期2301-2308,共8页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(82160099);贵州省科技计划项目(黔科合基础-ZK[2021]一般356);贵州省科技厅基础研究计划(黔科合基础-ZK[2022]一般465)。

摘　　要：目的 通过丙磺舒(PRB)抑制有机阴离子转运蛋白(OAT)1和3,观察马兜铃酸Ⅰ(AAⅠ)对大鼠肾小管上皮细胞的急性损伤,以探究AAⅠ进入肾小管上皮细胞的途径。方法雄性SD大鼠随机分为空白对照组、溶剂对照(PEG300)组、PRB(150 mg·kg^(-1))组、AAⅠ(80 mg·kg^(-1))组、AAⅠ(80 mg·kg^(-1))+PRB(150mg·kg^(-1))组,每2天ig给药1次,连续给药4次。观察大鼠肾脏指数;生化仪检测血清肌酐(CREA)和尿素氮(BUN)水平;苏木精-伊红(HE)染色观察肾脏组织病理学变化;免疫组化染色观察OAT1和OAT3在肾小管的组织定位和蛋白表达水平;免疫透射电镜观察OAT1和OAT3在肾小管上皮细胞的亚细胞定位和表达水平。结果 与溶剂对照组相比,AAⅠ组肾脏指数、CREA和BUN水平显著增加(P<0.01、0.001);AAⅠ+PRB组与AAⅠ组相比,肾脏指数、CREA和BUN水平显著降低(P<0.05、0.001)。HE染色结果显示,与溶剂对照组相比,AAⅠ组肾近端小管上皮细胞(PCTEC)出现空泡样变性、微绒毛脱落以及片状坏死脱落,而AAⅠ+PRB组与AAⅠ组相比,PCTEC空泡样变性率下降,无其他类型病理学变化;另外,肾远端小管上皮细胞(DCTEC)AAⅠ组与AAⅠ+PRB组呈现少量空泡样变性。免疫组化和电镜结果显示,OAT1主要在PCTEC基底膜侧表达,OAT3主要在DCTEC基底膜侧表达,且AAⅠ暴露后,与溶剂对照组比较,前者在近端小管(PCT)表达有下降趋势(P<0.05),后者在远端小管(DCT)表达有上升趋势(P<0.05)。结论 AAⅠ能够导致PCT和DCT的损伤,PRB抑制OAT1和OAT3后,能够改善肾脏功能,并减少肾小管上皮细胞的病理学损害;OAT1可能是AAⅠ进入PCTEC的主要通道,而OAT3则可能是其进入DCTEC的主要通道。Objective Utilizing probenecid(PRB)as an inhibitor of organic anion transporter(OAT)1 and 3,to investigate the acute injury of renal tubular epithelial cells in rats induced by aristolochic acid I(AAⅠ)and elucidate the pathway through which AAⅠenters renal tubular epithelial cells.Methods Male Sprague-Dawley rats were randomly assigned to five groups:the blank control group,the solvent control(PEG300)group,the PRB(150 mg·kg^(−1))group,the AAⅠ(80 mg·kg^(−1))group,and the AAⅠ+PRB(80 mg·kg^(−1)+150 mg·kg^(−1))group.Administer ig once every two days for four consecutive doses.Observed the renal organ index of rats,and biochemical analyzer detected serum creatinine(CREA)and urea nitrogen(UREA)levels.Hematoxylin-eosin(HE)staining was employed to examine the pathological alterations in renal tissue.Immunohistochemical(IHC)staining was conducted to assess the tissue localization and protein expression levels of OAT1 and OAT3 in renal tubules.Immunoelectron microscopy was utilized to investigate the subcellular localization and expression levels of OAT1 and OAT3 in renal tubular epithelial cells.Results Compared with the solvent control group,the renal index,CREA,and BUN levels in the AAⅠgroup were significantly increased(P<0.01,0.001);Compared with the AAⅠgroup,the renal index,CREA,and BUN levels were significantly reduced in the AAⅠ+PRB group(P<0.05,0.001).HE staining revealed cellular edema,necrotic shedding,microvillous loss in proximal convoluted tubule epithelial cells(PCTEC)in the AAⅠgroup(P<0.05).Conversely,in the AAⅠ+PRB group,the incidence of vacuolar degeneration in PCTEC decreased significantly(P<0.05)without other observed pathological changes.Additionally,a minor degree of vacuolar degeneration was observed in distal convoluted tubule epithelial cells(DCTEC)in both the AAⅠand AAⅠ+PRB groups.IHC and electron microscopy revealed that OAT1 was predominantly expressed in the basolateral membrane of PCTEC,while OAT3 was primarily expressed in the basolateral membrane of DCTEC.Fol

关 键 词：马兜铃酸I 有机阴离子转运蛋白1 有机阴离子转运蛋白3 丙磺舒 肾小管损伤 

分 类 号：R992[医药卫生—毒理学]