【摘要】：目的觀察Nrf2-ARE通路對大鼠心臟移植冷缺血再灌注損傷的影響及萊菔硫烷保護(hù)大鼠移植心臟的相關(guān)機(jī)制。方法1實(shí)驗(yàn)?zāi)Ｐ偷慕⒑头纸M:健康雄性Sprague-Dawley(SD)大鼠40只隨機(jī)分成3組:Sham組(假手術(shù)組,n=8),I/R組(缺血再灌注組,n=16),I/R+SFN組(萊菔硫烷預(yù)處理組,n=16)。Sham組:僅行開/關(guān)腹手術(shù),術(shù)前24h經(jīng)鼠尾靜脈注射等量的生理鹽水;I/R組:受體于移植前24h經(jīng)鼠尾靜脈注射等量的生理鹽水;I/R+SFN組:受體于移植前24h經(jīng)尾靜脈注射SFN2.5 mg/kg;其中I/R組和I/R+SFN組均將冷藏于4℃HTK液9 h的供心移植到受體大鼠腹腔,建立大鼠同種異體異位心臟移植模型,移植后24h兩組均將移植心臟取出;Sham組開/關(guān)腹24h后取自體心臟。2檢測指標(biāo):心肌酶學(xué):再灌注后6h、24h分別從受體鼠眼內(nèi)眥靜脈取血,檢測血清中乳酸脫氫酶(LDH)、肌酸激酶同工酶(CKMB)、肌鈣蛋白T(Tn T)的水平。采用硫代巴比妥酸法(TBARS)法、黃嘌呤氧化酶法分別檢測心肌組織中脂質(zhì)過氧化物(LPO)含量及超氧化物歧化酶(SOD)的活性。用HE染色法觀察各組心臟移植后供心心肌組織學(xué)變化。應(yīng)用免疫組織化學(xué)法(En Vision兩步法)及免疫蛋白印跡法觀察Nrf2、HO-1和NQO1在各組心臟移植后24h供心心肌組織的蛋白表達(dá)水平。結(jié)果心肌酶學(xué):與Sham組比較,I/R組,I/R+SFN組血清中LDH、CK-MB、Tn T含量明顯升高(P0.05);萊菔硫烷預(yù)處理后,缺血再灌注6h,與I/R組比較,I/R+SFN組血清中LDH、CK-MB、Tn T含量明顯降低(P0.05),再灌注24h,I/R+SFN組Tn T含量、CK-MB活性、LDH活性明顯降低(P0.05)。心肌組織中LPO含量及SOD的活性:與Sham組相比,I/R組和I/R+SFN組心肌組織SOD活性明顯降低(P0.05),而心肌組織LPO含量明顯升高(P0.05);與I/R組比較,I/R+SFN組心肌組織SOD活性未見明顯變化;而心肌組織LPO含量顯著降低(P0.05)。心肌組織學(xué):Sham組:心肌纖維排列整齊,結(jié)構(gòu)清楚,無心肌纖維的破壞,組織間隙無炎細(xì)胞滲出,胞核清晰。I/R組:心肌細(xì)胞結(jié)構(gòu)紊亂、較多心肌纖維斷裂、大量中性粒細(xì)胞浸潤;I/R+SFN組:心肌結(jié)構(gòu)較清楚,心肌纖維排列較整齊,心肌組織中心肌間質(zhì)散在中性粒細(xì)胞浸潤。心肌組織Nrf2及其下游分子HO-1和NQO1蛋白表達(dá):Sham組:心肌組織內(nèi)Nrf2蛋白幾乎不表達(dá)、HO-1蛋白弱表達(dá)、NQO1蛋白也是弱表達(dá)。與Sham組比較,I/R組心肌組織Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平顯著升高(P0.01);與I/R組比較,I/R+SFN組心肌組織Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平顯著升高(P0.01),但與Sham組比較,心肌組織Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平仍高于正常(P0.01)。結(jié)論1心臟移植冷缺血再灌注損傷可以激活Nrf2-ARE通路。2萊菔硫烷可能是通過激活Nrf2-ARE通路,上調(diào)下游基因HO-1、NQO1表達(dá),提高心肌細(xì)胞對氧化應(yīng)激的防御能力,對移植心臟冷缺血再灌注損傷起保護(hù)作用。
[Abstract]:Objective to investigate the effects of Nrf2-ARE pathway on cold ischemia reperfusion injury in rat heart transplantation and the protective mechanism of sulforaphane. Methods 1 Establishment and grouping of experimental models: forty healthy male Sprague-Dawley (SD) rats were randomly divided into 3 groups: group 1: sham-Sham group (sham group, n = 8), group I / R, group I / R SFN (preconditioning group with sulforaphane, group n 16) .Sham group: open / close abdominal surgery only. 24 hours before transplantation, the same amount of normal saline was injected into the tail vein of rats in the I / R group: the receptor group was injected with the same amount of normal saline intravenously 24 hours before transplantation: the receptor was injected with SFN 2.5 mg / kg through the tail vein 24 hours before transplantation, in which both the I / R group and the I / R / SFN group were treated with SFN 2.5 mg / kg. The donor heart was transplanted into the abdominal cavity of the recipient rats after being refrigerated in HTK solution at 4 鈩，

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