摘要
目的:探讨脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)信号通路介导莱菔硫烷(SFN)预处理对大鼠心肌冷缺血再灌注损伤(IRI)的作用,阐明其作用机制。方法:64只健康雄性SD大鼠随机分为冷IRI组、SFN组、LY(PI3K抑制剂)+冷IRI组和LY+SFN组,每组供、受体各8只。将冷藏于组氨酸-色氨酸-酮戊二酸盐液(HTK液)9h供心移植到受体大鼠的腹腔,建立同种大鼠异体异位心脏移植模型,术后24h取供心心肌组织,采用苏木精-伊红(HE)染色观察心肌组织形态表现,免疫组织化学法和Western blotting法检测心肌组织中丝氨酸/苏氨酸蛋白激酶,即蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、Bax、B细胞淋巴瘤/白血病-2(Bcl-2)的蛋白表达水平。结果:心肌组织形态表现,冷IRI组和LY+冷IRI组大鼠心肌组织损伤最严重,SFN组心肌组织损伤最轻,LY+SFN组心肌组织损伤介于冷IRI组和SFN组之间。免疫组织化学法和Western blotting法,与冷IRI组比较,SFN组p-Akt蛋白表达水平明显升高(P<0.05),Bcl-2蛋白表达水平升高(P<0.05),而Bax蛋白表达水平降低(P<0.05);应用阻滞剂LY294002后,与LY+冷IRI组比较,LY+SFN组p-Akt蛋白表达水平差异无统计学意义(P>0.05),但Bcl-2蛋白表达水平仍升高(P<0.05),Bax蛋白表达水平仍降低(P<0.05),Bcl-2/Bax比值升高(P<0.05)。结论:SFN可能通过PI3K/Akt信号通路减轻心脏移植心肌冷IRI。
Objective:To explore the effect of sulforaphane(SFN)preconditioning on the cold myocardial ischemia-reperfusion injury(IRI)in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley(SD)rats were randomly divided into cold IRI group,SFN group,LY(LY294002) +cold IRI group,and LY+SFN group(n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 hafter reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2protein in SFN group were increased(P〈0.05),and the expression level of Bax protein was decreased(P〈0.05).After treatment of blockage LY294002,compared with LY+cold IRI group,the expression level of p-Akt protein in LY+SFN group was not statistically significant(P〉0.05),the expression level of Bcl2 protein was increased(P〈0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased(P〈0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.
出处
《吉林大学学报(医学版)》
CAS
CSCD
北大核心
2017年第5期918-922,I0005,共6页
Journal of Jilin University:Medicine Edition
基金
福建省科技厅自然科学基金资助课题(2016D014)
福建省卫计委医学创新项目资助课题(2015-CXB-47)