氧化槐定碱通过Akt/Nrf2/HO-1和Akt/GSK3β信号通路对急性肾损伤的保护作用  

Protective effect of oxysophoridine from acute kidney injury by Akt/Nrf2/HO-1 and Akt/GSK3β signaling pathways

作　　者：王文文[1] 张敏[2] 

机构地区：[1]浙江大学医学院附属妇产科医院病理科,杭州310006 [2]浙江中医药大学附属第一医院病理科

出　　处：《山西医科大学学报》2017年第10期992-998,共7页Journal of Shanxi Medical University

摘　　要：目的探讨氧化槐定碱(oxysophoridine,OSR)对小鼠肾缺血再灌注(ischemia/reperfusion,I/R)诱导的急性肾损伤的保护作用及其机制。方法 C57/BL6小鼠30只随机分为假手术组(sham组)、模型组(I/R组)和给药组(I/R+OSR组)。I/R+OSR组预给药5 d[250 mg/(kg·d),腹腔注射],sham组和I/R组给予相同体积的生理盐水。第5天给药后1 h开始利用左侧肾切除、右侧肾蒂缺血30 min再灌注24 h的方法构建缺血再灌注损伤模型。下腔静脉取血并离心取血清,用自动生化仪检测肌酐和尿素氮,试剂盒检测丙二醛的生成及超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)的活性,ELISA法检测肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β),HE染色法检测肾脏组织形态学,Western blot法检测蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、细胞内核因子E2相关因子2(Nrf2)、血红素加氧酶(HO-1)、糖原合酶激酶-3β(GSK3β)、磷酸化GSK3β(p-GSK3β)、半胱天冬酶-3(pro-caspase-3)、活化的半胱天冬酶-3(cleaved caspase-3)、B淋巴细胞瘤-2(Bcl-2)的表达。结果与sham组比较,I/R组小鼠肌酐、尿素氮、丙二醛、TNF-α和IL-1β显著升高(P<0.05),SOD、CAT的活性显著降低(P<0.05),p-Akt、Nrf2、HO-1、p-GSK3β、Bcl-2、pro-caspase-3蛋白表达显著降低(P<0.05),而促凋亡蛋白cleaved caspase-3表达显著升高(P<0.05)。与I/R组比较,OSR组小鼠肌酐、尿素氮、丙二醛、TNF-α和IL-1β显著降低(P<0.05),SOD、CAT的活性显著增加(P<0.05),p-Akt、Nrf2、HO-1、p-GSK3β、Bcl-2、pro-caspase-3蛋白表达显著升高(P<0.05),促凋亡蛋白cleaved caspase-3表达显著降低(P<0.05)。与sham组比较,HE染色结果显示I/R组肾损伤显著加重,而给药后可显著改善急性肾损伤。结论 OSR通过Akt/Nrf2/HO-1和Akt/GSK3β信号通路及抗炎和抗凋亡相关信号通路机制对肾缺血再灌注诱导的急性肾损伤起到保护作用。Objective To investigate the protective effects of oxysophoridine（ OSR） from renal ischemia reperfusion-induced acute kidney injury in mice and its possible mechanism. Methods C57/BL6 mice were randomly divided into 3 groups： sham-operated group（sham group）,model group（I/R group）,OSR group（I/R ＋ OSR）. The mice in OSR group were pretreated with OSR for 5 d before inducing the I/R injury model（250 mg/kg,once a day,intraperitoneal injection）. The mice in sham group and I/R group were given the same volume of physiological saline. One hour after the last administration,the right renal pedicle was clamped for 30 min and left nephrectomy was performed to produce I/R injury model in mice. Blood samples were collected from the inferior vena cava and centrifuged. Serum creatinine and blood urea nitrogen were measured by an automatic biochemical analyzer. Malondialdehyde concentration and enzymatic activity of superoxide dismutase（ SOD） and catalase（ CAT） were measured by Kits respectively. Tumor necrosis factor alpha（ TNF-α） and interleukin-1 beta（ IL-1β） were detected by ELISA Kits. The histological structure changes of renal tissues were observed under microscope. The expression levels of protein kinase B（Akt）,phosphorylated Akt（ p-Akt）,nuclear factor erythroid 2-related factor 2（ Nrf2）,heme oxygenase（ HO-1）,glycogen synthase kinase 3 beta（ GSK3β）,phosphorylation GSK3β（ p-GSK3β）,pro-caspase-3,cleaved caspase-3,B-cell lymphoma-2（ Bcl-2） were detected by Western blot. Results Compared with sham group,serum creatinine,blood urea nitrogen,malondialdehyde,TNF-α and IL-1β increased significantly in I/R group（ P 〈 0. 05）,SOD and CAT decreased significantly in I/R group（ P 〈 0. 05）,and the expression of p-Akt,Nrf2,HO-1,p-GSK3β,Bcl-2,pro-caspase-3 reduced and cleaved caspase-3 increased significantly in I/R group（ P 〈 0. 05）. Compared with model group,serum creatinine,blood urea nitrogen,malondialdehyde,TNF-α and IL-1β decreased signi

关 键 词：氧化槐定碱 肾缺血再灌注 细胞内核因子E2相关因子2 血红素加氧酶 糖原合酶激酶-3Β 

分 类 号：R363[医药卫生—病理学] R692[医药卫生—基础医学]