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

发布时间：2018-07-17 17:37

【摘要】：目的观察Nrf2-ARE通路对大鼠心脏移植冷缺血再灌注损伤的影响及莱菔硫烷保护大鼠移植心脏的相关机制。方法1实验模型的建立和分组:健康雄性Sprague-Dawley(SD)大鼠40只随机分成3组:Sham组(假手术组,n=8),I/R组(缺血再灌注组,n=16),I/R+SFN组(莱菔硫烷预处理组,n=16)。Sham组:仅行开/关腹手术,术前24h经鼠尾静脉注射等量的生理盐水;I/R组:受体于移植前24h经鼠尾静脉注射等量的生理盐水;I/R+SFN组:受体于移植前24h经尾静脉注射SFN2.5 mg/kg;其中I/R组和I/R+SFN组均将冷藏于4℃HTK液9 h的供心移植到受体大鼠腹腔,建立大鼠同种异体异位心脏移植模型,移植后24h两组均将移植心脏取出;Sham组开/关腹24h后取自体心脏。2检测指标:心肌酶学:再灌注后6h、24h分别从受体鼠眼内眦静脉取血,检测血清中乳酸脱氢酶(LDH)、肌酸激酶同工酶(CKMB)、肌钙蛋白T(Tn T)的水平。采用硫代巴比妥酸法(TBARS)法、黄嘌呤氧化酶法分别检测心肌组织中脂质过氧化物(LPO)含量及超氧化物歧化酶(SOD)的活性。用HE染色法观察各组心脏移植后供心心肌组织学变化。应用免疫组织化学法(En Vision两步法)及免疫蛋白印迹法观察Nrf2、HO-1和NQO1在各组心脏移植后24h供心心肌组织的蛋白表达水平。结果心肌酶学:与Sham组比较,I/R组,I/R+SFN组血清中LDH、CK-MB、Tn T含量明显升高(P0.05);莱菔硫烷预处理后,缺血再灌注6h,与I/R组比较,I/R+SFN组血清中LDH、CK-MB、Tn T含量明显降低(P0.05),再灌注24h,I/R+SFN组Tn T含量、CK-MB活性、LDH活性明显降低(P0.05)。心肌组织中LPO含量及SOD的活性:与Sham组相比,I/R组和I/R+SFN组心肌组织SOD活性明显降低(P0.05),而心肌组织LPO含量明显升高(P0.05);与I/R组比较,I/R+SFN组心肌组织SOD活性未见明显变化;而心肌组织LPO含量显著降低(P0.05)。心肌组织学:Sham组:心肌纤维排列整齐,结构清楚,无心肌纤维的破坏,组织间隙无炎细胞渗出,胞核清晰。I/R组:心肌细胞结构紊乱、较多心肌纤维断裂、大量中性粒细胞浸润;I/R+SFN组:心肌结构较清楚,心肌纤维排列较整齐,心肌组织中心肌间质散在中性粒细胞浸润。心肌组织Nrf2及其下游分子HO-1和NQO1蛋白表达:Sham组:心肌组织内Nrf2蛋白几乎不表达、HO-1蛋白弱表达、NQO1蛋白也是弱表达。与Sham组比较,I/R组心肌组织Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平显著升高(P0.01);与I/R组比较,I/R+SFN组心肌组织Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平显著升高(P0.01),但与Sham组比较,心肌组织Nrf2核蛋白、Nrf2、HO-1和NQO1蛋白水平仍高于正常(P0.01)。结论1心脏移植冷缺血再灌注损伤可以激活Nrf2-ARE通路。2莱菔硫烷可能是通过激活Nrf2-ARE通路,上调下游基因HO-1、NQO1表达,提高心肌细胞对氧化应激的防御能力,对移植心脏冷缺血再灌注损伤起保护作用。
[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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