丹酚酸B基于Nrf2-Gpx4通路介导的铁死亡途径改善单侧输尿管梗阻大鼠肾脏间质纤维化  

Salvianolic acid B attenuates renal interstitial fibrosis in rats with unilateral ureteral obstruction via Nrf2-Gpx4 pathway-mediated ferroptosis

作　　者：孙源博 宋嫣然 王诗琪 孙丽欣 李桂芹 SUN Yuanbo;SONG Yanran;WANG Shiqi;SUN Lixin;LI Guiqin(Department of Nephrology,Hongqi Hospital Affiliated to Mudanjiang Medical College,Mudanjiang,Heilongjiang Province,157000,China;Hemodialysis Room,Hongqi Hospital Affiliated to Mudanjiang Medical College,Mudanjiang,Heilongjiang Province,157000,China)

机构地区：[1]牡丹江医学院附属红旗医院:肾脏内科,黑龙江牡丹江157000 [2]牡丹江医学院附属红旗医院:血液透析室,黑龙江牡丹江157000

出　　处：《陆军军医大学学报》2022年第10期1018-1024,共7页Journal of Army Medical University

基　　金：黑龙江省青年创新人才培养计划项目(UNPYSCT-2018120);黑龙江省省属高等学校基本科研业务费科研项目(2019-KYYWFMY-0047);“红旗科研基金”科技项目(2018HQ-05)。

摘　　要：目的基于核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)-谷胱甘肽过氧化物酶4(glutathione peroxidase 4,Gpx4)通路介导的铁死亡途径,探究丹酚酸B对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾脏间质纤维化(renal interstitial fibrosis,RIF)的改善作用。方法采用随机数字表法将40只大鼠分为假手术组、模型组、丹酚酸B组、丹酚酸B+Nrf2抑制剂组,每组10只。除假手术组外,其余各组建立UUO模型,假手术组除不结扎、剪断输尿管外,其余操作相同。丹酚酸B组灌胃100 mg·kg^(-1)·d^(-1)丹酚酸B,丹酚酸B+Nrf2抑制剂组在丹酚酸B组基础上腹腔注射Nrf2抑制剂ML38530 mg·kg^(-1)·d^(-1),假手术组、模型组灌胃等体积无菌生理盐水,连续9 d。HE染色和Masson染色检测肾组织形态和纤维化情况;试剂盒检测肾脏组织中Fe^(2+)、丙二醛(maleicdialdehyde,MDA)含量及超氧化物歧化酶(superoxide dismutase,SOD)活性;实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)和蛋白质免疫印迹检测肾脏组织中Ⅰ型胶原(Collagen-Ⅰ,CoL-Ⅰ)、Ⅲ型胶原(Collagen-Ⅲ,CoL-Ⅲ)、Nrf2、Gpx4的mRNA和蛋白水平。结果与假手术组相比,模型组肾脏组织中Fe^(2+)、MDA含量,CoL-Ⅰ、CoL-Ⅲ的mRNA和蛋白水平升高(P<0.05),血清中SOD活性降低(P<0.05),肾脏组织中Nrf2、Gpx4的mRNA和蛋白水平降低(P<0.05);与模型组相比,丹酚酸B组肾脏组织中Fe^(2+)、MDA含量,CoL-Ⅰ、CoL-Ⅲ的mRNA和蛋白水平降低(P<0.05),血清中SOD活性,肾脏组织中Nrf2、Gpx4的mRNA和蛋白水平升高(P<0.05);与丹酚酸B组相比,丹酚酸B+Nrf2抑制剂组肾脏组织中Fe^(2+)、MDA含量,CoL-Ⅰ、CoL-Ⅲ的mRNA和蛋白水平升高(P<0.05),血清中SOD活性,肾脏组织中Nrf2、Gpx4的mRNA和蛋白水平降低(P<0.05)。结论丹酚酸B能够激活Nrf2-Gpx4通路进而抑制铁死亡实现对UUO大鼠RIF的缓解。ObjectiveTo determine the improving effect of salvianolic acid B on renal interstitial fibrosis(RIF)of rats with unilateral ureteral obstruction(UUO)based on ferroptosis mediated by nuclear factor erythroid 2-related factor 2(Nrf2)-glutathione peroxidase 4(Gpx4)pathway.MethodsA total of 40 rats were divided into sham operation group,model group,salvianolic acid B group,and salvianolic acid B+Nrf2 inhibitor group,with 10 rats in each group.UUO model was established in the latter 3 groups,and those of the sham operation group received the same operation except for not ligating and cutting the ureter.Salvianolic acid B of 100 mg·kg^(-1)·d^(-1)and Nrf2 inhibitor ML385 of 30 mg·kg^(-1)·d^(-1)were intragastrically administered to the corresponding groups,and sterile normal saline of equal volume was given to the sham operation group and model group,for 9 consecutive days.Hematoxylin-eosin(HE)staining and Masson staining were used to detect the morphology and fibrosis in renal tissues.The contents of Fe^(2+),maleicdialdehyde(MDA)and activity of superoxide dismutase(SOD)in renal tissues were detected with corresponding kits.Quantitative real-time PCR(RT-qPCR)and Western blotting were used to detect the mRNA and protein levels of Collagen-Ⅰ(CoL-Ⅰ),Collagen-Ⅲ(CoL-Ⅲ),Nrf2,and Gpx4 in the renal tissues.ResultsCompared with the sham operation group,the contents of Fe^(2+)and MDA and the mRNA and protein levels of CoL-Ⅰand CoL-Ⅲin the renal tissues were increased(P<0.05),and the serum SOD activity and the mRNA and protein levels of Nrf2 were decreased(P<0.05)in the model group.While the contents of Fe^(2+)and MDA and the mRNA and protein levels of CoL-Ⅰand CoL-Ⅲwere decreased(P<0.05),and the SOD activity and the levels of Nrf2 and Gpx4 were increased(P<0.05)in the salvianolic acid B group than the model group.The treatment of salvianolic acid B+Nrf2 inhibitor increased the contents of Fe^(2+)and MDA and the mRNA and protein levels of CoL-Ⅰand CoL-Ⅲ(P<0.05),and reduced the SOD activity and the levels of

关 键 词：肾脏间质纤维化 单侧输尿管梗阻 丹酚酸B 核因子E2相关因子2-谷胱甘肽过氧化物酶4通路 

分 类 号：R285.5[医药卫生—中药学] R692[医药卫生—中医学]