PI3K/Akt/Nrf2信号通路在白藜芦醇预处理对糖尿病大鼠心肌保护效应中的作用  

Role of PI3K/Akt/Nrf2 signaling pathway in resveratrol preconditioning-induced cardioprotection in diabetic rats

作　　者：徐桂萍[1] 付鹃 赵萱 王晓丽[1] Xu Guiping;Fu Juan;Zhao Xuan;Wang Xiaoli(Department of Anesthesiology,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,China;Graduate School,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆维吾尔自治区人民医院麻醉科,乌鲁木齐830000 [2]新疆医科大学研究生学院,乌鲁木齐830011

出　　处：《中华麻醉学杂志》2019年第10期1189-1193,共5页Chinese Journal of Anesthesiology

基　　金：新疆维吾尔自治区自然科学基金(2018D01C110)。

摘　　要：目的评价磷脂酰肌醇-3-激酶/蛋白激酶B/核因子E2相关因子2(PI3K/Akt/Nrf2)信号通路在白藜芦醇预处理对糖尿病大鼠心肌保护效应中的作用。方法清洁级健康雄性SD大鼠,2~3月龄,体重240~280 g,腹腔注射链脲佐菌素30 mg/kg,每天1次,连续7 d,制备糖尿病模型。取糖尿病大鼠40只,按照随机数字表法分为4组(n=10):假手术组(S组)、心肌缺血再灌注组(I/R组)、白藜芦醇+心肌缺血再灌注组(Res+I/R组)、PI3K抑制剂LY294002+白藜芦醇+心肌缺血再灌注组(LY+Res+I/R组)。采用结扎冠状动脉左前降支30 min再灌注120 min的方法制备心肌缺血再灌注损伤模型。Res+I/R组和LY+Res+I/R组术前1周腹腔注射白藜芦醇20 mg/kg,1次/d,连续7 d;LY+Res+I/R组术前30 min腹腔注射LY2940021.5 mg/kg。再灌注120 min时,取血样,采用ELISA法测定血清CK-MB和LDH水平;取缺血心肌组织,采用ELISA法测定SOD、谷胱甘肽(GSH)和MDA水平,采用Western blot法检测Akt、磷酸化Akt(p-Akt)、糖原合成酶激酶3β(GSK3β)、磷酸化GSK3β(p-GSK3β)和核因子E2相关因子2(Nrf2)的表达。结果与S组比较,I/R组血清CK-MB和LDH浓度升高,SOD和GSH水平降低,MDA水平升高,p-Akt、p-GSK3β表达下调(P<0.05);与I/R组比较,Res+I/R组血清CK-MB和LDH浓度降低,SOD、GSH水平升高,MDA水平降低,p-Akt、p-GSK3β和Nrf2表达上调(P<0.05);与Res+I/R组比较,LY+Res+I/R组血清CK-MB和LDH浓度升高,SOD、GSH水平降低,MDA水平升高,p-Akt、p-GSK3β和Nrf2表达下调(P<0.05)。结论白藜芦醇预处理对糖尿病大鼠心肌保护效应的机制与激活PI3K/Akt/Nrf2信号通路,抑制氧化应激反应有关。Objective To evaluate the role of phosphatidylinositol-3-kinase/protein kinase B/nuclear factor erythroid 2-related factor 2(PI3K/Akt/Nrf2)signaling pathway in resveratrol preconditioning-induced cardioprotection in diabetic rats.Methods Clean-grade healthy male Sprague-Dawley rats,aged 2-3 months,weighing 240-280 g,were used in the study.The diabetes model was established by intraperitoneal injection of streptozotocin 30 mg/kg once a day for 7 consecutive days.Forty diabetic rats were selected and divided into 4 groups(n=10 each)according to the random number table method:sham operation group(S group),myocardial ischemia-reperfusion(I/R)group(I/R group),resveratrol plus myocardial I/R group(Res+I/R group),and PI3K inhibitor LY294002 plus resveratrol plus myocardial I/R group(LY+Res+I/R group).The myocardial I/R injury model was established by ligating the left anterior descending coronary artery for 30 min followed by 120-min reperfusion in anesthetized rats.Resveratrol 20 mg/kg was intraperitoneally injected once a day for 7 consecutive days at one week before surgery in Res+I/R and LY+Res+I/R groups.LY2940021.5 mg/kg was intraperitoneally injected at 30 min before operation in LY+Res+I/R group.After 120 min of reperfusion,blood samples were taken for determination of serum creatine kinase-MB(CK-MB)and lactic dehydrogenase(LDH)concentrations(by enzyme-linked immunosorbent assay),and myocardial tissues at the ischemic area were obtained for determination of superoxide dismutase(SOD),glutathione(GSH)and malondialdehyde(MDA)levels(by enzyme-linked immunosorbent assay)and expression of Akt,phosphorylated Akt(p-Akt),glycogen synthase kinase 3β(GSK3β),phosphorylated GSK3β(p-GSK3β)and Nrf2(by Western blot).Results Compared with group S,serum CK-MB and LDH concentrations were significantly increased,the SOD and GSH levels were decreased,the MDA level was increased,and the expression of p-Akt and p-GSK3βwas down-regulated in I/R group(P<0.05).Compared with I/R group,serum CK-MB and LDH concentrations were significa

关 键 词：心肌再灌注损伤 糖尿病 酚类 1-磷脂酰肌醇3-激酶 蛋白质丝氨酸苏氨酸激酶 NF-E2相关因子2 

分 类 号：R285.5[医药卫生—中药学]