莱菔硫烷通过Nrf2-ARE通路减轻大鼠移植心脏冷缺血再灌注损伤

摘要

Objective To discuss whether sulforaphane (SFN) can reduce cold ischemia-reperfusion injury through Nrf2-ARE pathway in rats' heart transplantation.Methods 40 health Male Sprague-Dawley(SD) rats were randomly divided into 3 groups:sham group:n =8,I/R group--ischemia reperfusion group,n =16,I/R + SFN group-sulforaphane preconditioning group,n =16.Donnor rats' hearts stored in 4℃ HTK solution for 9 hours were transplanted into recipient rats intraperitoneal,and then rat heterotopic heart transplantation model was performed in I/R and 1/R + SFN groups.Autologous heart and graft were take out 24 hours after heart transplantation in sham group,IRI group,and IRI + SFN group.Cardiac histology and nuclear factor-related factor 2(Nrf2),heme oxygenase 1 (HO-1) and quinone oxidoreductase 1 (NQO1) protein were observed using hematoxylin-eosin staining,immunohistochemistry and western blot method in each group.Results I/R pretreatment group myocardial tissue damage mitigation,myocardial tissue Nrf2,HO-1,NQO1 protein levels were significantly increased(P ＜0.01) compared with ischemia reperfusion group.However,myocardial histology and myocardial tissue Nrf2 nuclear protein,Nrf2,HO-1 and NQO1 protein levels still higher than normal(P ＜ 0.01) compared with sham group.Conclusion Sulforaphane improves myocardial cells against oxidative stress defense capability from heart transplantation cold ischemia-reperfusion injury by activating Nrf2-ARE pathway regulated downstream protein HO-1,NQO1 expression.%目的 探讨莱菔硫烷(SFN)是否通过核因子相关因子2-抗氧化反应元件(Nrf2-ARE)通路对抗大鼠心脏移植冷缺血再灌注损伤.方法 健康雄性Sprague-Dawley(SD)大鼠40只随机分成3组:假手术组(Sham组,n=8),缺血再灌注组(I/R组,n=16),莱菔硫烷预处理组(I/R+ SFN组,n=16).建立大鼠异体异位心脏移植模型,将冷藏于4℃ HTK液9h的供心移植到I/R组和I/R+ SFN组受体大鼠腹腔,移植后24h取出移植心脏.Sham组开/关腹24 h后取出自体心脏.应用HE染色法、免疫组织化学法及western blot方法观察心肌组织学、核因子相关因子2(Nrf2)、血红素加氧酶1(HO-1)、醌氧化还原酶1(NQO1)的蛋白表达水平.结果 与I/R组比较,I/R+ SFN组心肌组织损伤减轻,心肌组织Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平显著升高(P＜0.01).与Sham组比较,I/R+ SFN组心肌组织学和心肌组织Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平仍高于正常(P＜0.01).结论 莱菔硫烷通过激活Nrf2-ARE通路,上调下游蛋白HO-1、NQO1表达,提高心肌细胞对氧化应激的防御能力,对抗移植心脏冷缺血再灌注损伤.