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

Sulforaphane reduces cold myocardial ischemia reperfusion injury through Nrf2-ARE pathway in rat heart transplantation

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24h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NQ01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。

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. Dounor 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 I/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 ） , heine oxygenase 1 （ HO-1 ） and quinone oxidoreductase 1 （ NQOI ） 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, myoeardial 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.