磷霉素对万古霉素致肾损伤模型大鼠Keap1-Nrf2/ARE通路的影响  

Effect of Fosfomycin on Keap1-Nrf2/ARE Pathway in Vancomycin-induced Kidney Injury of Model Rats

作　　者：黄晓青[1] 黄丽蓓 杨斌[2] 林子杰 梁雪 何庆林 丘岳[1] HUANG Xiaoqing;HUANG Libei;YANG Bin;LIN Zijie;LIANG Xue;HE Qinglin;QIU Yue(Department of Pharmacy,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;College of Pharmacy,Guangxi Medical University,Nanning 530021,China;The First Clinical Medical College,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院药学部,南宁530021 [2]广西医科大学药学院,南宁530021 [3]广西医科大学第一临床医学院,南宁530021

出　　处：《医药导报》2023年第1期12-16,共5页Herald of Medicine

基　　金：广西教育厅高校中青年教师科研基础能力提升项目(2019KY0131);广西壮族自治区卫生健康委课题(Z20190075);大学生创新创业项目(国家级:201910598039)。

摘　　要：目的探讨磷霉素对万古霉素致肾损伤模型大鼠的保护作用及其机制。方法SD大鼠60只,随机分为模型对照组、正常对照组、磷霉素组和万古霉素联合磷霉素小、中、大剂量组(联合-L、联合-M、联合-H组),每组10只。正常对照组给予等剂量0.9%氯化钠注射液,模型对照组给予万古霉素0.2 g·kg^(-1),磷霉素组给予磷霉素0.25 g·kg^(-1),联合-L组、联合-M组、联合-H组分别给予磷霉素0.125,0.25,0.5 g·kg^(-1),30 min后再给予万古霉素0.2 g·kg^(-1),均腹腔注射,连续21 d。酶联免疫吸附实验(ELISA)测定大鼠尿N-乙酰-β-D-氨基葡萄糖苷酶(NAG)、肾损伤因子1(KIM-1)含量,考马斯亮蓝法测定24 h尿蛋白,反转录-聚合酶链反应(RT-PCR)检测肾组织Kelch样环氧氯丙烷相关蛋白-1(Keap1)、核因子E2相关因子(Nrf2)、抗氧化反应元件(ARE)信号通路Nrf2、Keap1及下游靶蛋白谷胱甘肽过氧化物酶-1(GPX-1)mRNA表达。结果与正常对照组比较,模型对照组24 h尿蛋白、NAG、KIM-1水平显著升高(P<0.05),肾组织Nrf2 mRNA表达升高(P<0.01),Keap1几乎无变化,GPX-1 mRNA表达下降(P<0.01);联合-L组、联合-M组Nrf2、Keap1 mRNA表达明显升高(P<0.01),联合给药组GPX-1 mRNA表达均显著升高(P<0.01)。与模型对照组比较,联合-M组、联合-H组24 h尿蛋白、NAG、KIM-1水平显著下降(P<0.01),且与磷霉素呈剂量依赖性。结论万古霉素可通过影响肾组织Nrf2表达引起肾损伤。磷霉素可能通过激活机体抵抗外界氧化刺激的防御性转导Keap1-Nrf2/ARE信号通路改善万古霉素引起的肾损伤。Objective To explore the protective effect and mechanism of fosfomycin on vancomycin-induced renal injury of model rats.Methods A total of 60 healthy SD rats were randomly divided into the normal control group,model control group,vancomycin with high,medium,and low dose fosfomycin groups(recorded as combined-L,combined-M,and combined-H group),10 animals in each group.Each rat in the normal control group was injected intraperitoneally(IP)with the same dose of 0.9%sodium chloride solution.The rats in the model control group were injected with vancomycin(0.2 g·kg^(-1)),in the fosfomycin group was injected with fosfomycin(0.25 g·kg^(-1)).In combination regimens,rats were injected with fosfomycin at a dose of 0.125,0.25,and 0.5 g·kg^(-1)respectively in combined-L,combined-M,and combined-H group,and 30 minutes later rats were injected with vancomycin(0.2 g·kg^(-1)).The drug was administered for 21 days continuously.The contents of NAG and KIM-1 in urine were determined by enzyme-linked immunosorbent assay(ELISA),proteinuria(Upro/24 h)by Coomassie brilliant blue staining method,and the expressions of Nrf2,Keap1,and the target protein glutathione peroxidase1(GPX-1)downstream of the Keap1-Nrf2/ARE pathway were detected by RT-PCR.Results Compared with the model control group,the levels of Upro/24h,NAG,and KIM-1 in the urine of the model control group were significantly higher(P<0.05),the mRNA expression of Nrf2 was significantly increased(P<0.01),Keap1 was almost unchanged,and the mRNA expression of GPX-1 was decreased(P<0.01)in model group.Expressions of Nrf2 and Keap1 mRNA in the combined-L and the combined-M were significantly increased(P<0.01),and the expression of GPX-1 mRNA in three groups of the combination regimen was significantly increased(P<0.01).Compared with the model control group,the level of Upro/24 h,NAG,and KIM-1 in combined-M and combined-H groups decreased significantly(P<0.01).With the increase in fosfomycin dose,it showed a certain dose dependence.Conclusion Vancomycin may cause renal injury by af

关 键 词：磷霉素 万古霉素 肾损伤 氧化应激 

分 类 号：R978.19[医药卫生—药品] R965[医药卫生—药学]