摘要
目的探讨二甲双胍能否减轻肾缺血再灌注损伤(I/R)后内质网(ER)应激、自噬作用和炎性反应。方法健康清洁级Wistar大鼠分为3组(n=6),即行假手术(Sham组)、双侧缺血1h再灌注2h(I/R组),缺血前腹腔注射二甲双胍0.125mg/(kg·d)14天(metformin组)。采用全自动生化分析仪测定血浆样品中肌酐(Cr)和乳酸脱氢酶(LDH)含量,通过紫外可见分光度计分别测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)、还原型谷胱甘肽(GSH)、丙二醛(MDA)含量;蛋白印迹分析GRP 78、CHOP、p-Eif2-α、Beclin-1、β-actin、XBP-1、ATF-6α、LAMP-2蛋白表达;RT-PCR检测IL-1β、IL-6和MCP-1表达;HE法进行组织学评估。结果二甲双胍可增强肾功能,减少细胞溶解(P<0.05)。与I/R组比较,显著提高了抗氧化酶(SOD、CAT、GPX)活性和GSH水平(P<0.05)。二甲双胍处理显著降低ER应激,这一作用通过下调GRP78、ATF-6、p-eIF-2α、XPB-1和CHOP得以证实。二甲双胍预处理的大鼠显示自噬参数(Beclin-1和LAMP-2)低表达,炎性因子(IL-1β、IL-6和MCP-1)减少(P<0.05)。结论二甲双胍对肾I/R具有保护作用,在I/R后减轻ER通路和自噬过程。
Objective To investigate whether metformin can reduce endoplasmic reticulum(ER)stress,autophagy and inflammatory response after renal ischemia-reperfusion injury(I/R).Methods Wistar rats were divided into three groups(n=6):sham operation(Sham group),bilateral ischemia 1h reperfusion for 2h(I/R group),and intraperitoneal injection of metformin 0.125mg/(kg·d)for 14 days(metformin group).The content of creatinine(Cr)and lactate dehydrogenase(LDH)in plasma samples was determined by automatic biochemical analyzer.Superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GPx),reduced glutathione(GSH)and malondialdehyde(MDA)content were determined by UV-Vis spectrophotometer.Western blot was used to analysis GRP 78,CHOP,p-Eif2-α,Beclin-1,β-actin,XBP-1,ATF-6α,LAMP-2 protein expression.RT-PCR was used to detect IL-1β,IL-6 and MCP-1 expression.HE method was used to test histological evaluation.Results Metformin enhanced renal function and decreased cell lysis(P<0.05).Compared with the I/R group,the activities of antioxidant enzymes(SOD,CAT,GPX)and GSH levels were significantly increased(P<0.05).Metformin treatment significantly reduced ER stress,which was confirmed by down-regulation of GRP78,ATF-6,p-eIF-2α,XPB-1 and CHOP.Metformin-pretreated rats showed low expression of autophagy parameters(Beclin-1 and LAMP-2)and decreased inflammatory factors(IL-1β,IL-6 and MCP-1)(P<0.05).Conclusion Metformin has a protective effect on renal I/R,and reduces the ER pathway and autophagy after I/R.
作者
金领微
潘敏
叶菡洋
陈琰
叶白如
郑育
潘殊方
施珍
张静
Jin Lingwei;Pan Min;Ye Hanyang(Department of Nephrology,The Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University,Zhejiang 325027,China)
出处
《医学研究杂志》
2020年第1期111-117,共7页
Journal of Medical Research
基金
浙江省医药卫生科技项目(2020358611)
浙江省温州市科技计划项目(Y20180499,Y20190063)
贺林院士新医学临床转化工作站科研基金资助项目。
关键词
自噬
内质网
炎症
缺血再灌注
氧化应激
二甲双胍
Autophagy
Endoplasmic reticulum
Inflammation
Ischemia-reperfusion
Oxidative stress
Metformin
作者简介
通讯作者:张静,电子信箱:23293313@qq.com。
