磷脂酰肌醇3激酶／丝苏氨酸蛋白激酶信号转导通路在芬太尼后处理和远隔缺血后处理心肌保护中的作用

Roles of phosphoinositide 3 kinase/serine-threonine kinase signal pathway in cardioprotection of fentanyl postconditioning and limb remote postconditioning

目的在心肌缺血，再灌注损伤（ischemia／reperfusioninjury，I／RD大鼠探讨磷脂酰肌醇3激酶／丝苏氨酸蛋白激酶（phosphoinositide3kinase／serine-threoninekinase，P13K／Akt）信号转导通路在芬太尼后处理和远隔缺血后处理心肌保护中的作用。方法将32只成年雄性sD大鼠（体重250g-350g）麻醉后，采用计算机产生的随机数随机分为4组：对照组（c组）、芬太尼后处理组（F组）、肢体远隔缺血后处理组（R组）及联合应用芬太尼后处理和肢体远隔缺血后处理组（F_R组）。在结扎大鼠冠状动脉左前降支（1eftanteriordescendingcoronaryartery，LAD）30min造成局部心肌缺血后，开放心肌再灌注60min建立大鼠心肌I／RI模型。采用SABioscience公司功能分类基因芯片和免疫蛋白印迹分析法检测再灌注60min后缺血区心肌内与P13K／Akt相关基因的表达和磷酸化Akt蛋白的表达情况。结果利用基因芯片检测的与P13K／Akt相关的基因中，与C组比较，F组共有9个基因的表达显著上调，而R组仅2个基因的表达显著上调；但F-R组共有33个基因的表达较C组显著上调。蛋白印记分析结果显示，与C组比较，F组、R组和F-R组心肌标本内磷酸化Akt蛋白表达量均增高；而与F组和R组比较，F-R组心肌标本内磷酸化Akt蛋白表达量进一步增高。结论联合应用芬太尼后处理和肢体远隔缺血后处理可明显增强P13K／Akt信号转导通路激活。

Objective To assess the roles of phosphoinositide 3 kinase/serine-threonine kinase （PI3K/Akt） signal pathway in cardioprotection of fentanyl postconditioning and limb remote postconditioning in an in vivo rat model with myocardial ischemia reperfusion injury. Methods Thirty-two anesthetized male SD rats （weighed 250 g-350 g） were randomly allocated into the four groups： group C （control）, group F （fentanyl postconditioning）, group R （RIPOC）, and group F-R （combined fentanyl postconditioning and RIPOC）. All rats were treated with left anterior descending coronary artery（LAD） occluded for 30 rain, followed by a 60-rain reperfusion （LAD open） in vivo. The Olingo PI3k/Akt signaling pathway microarray （SA Bioscience）and Western-blot technique were used to distinguish the expression of genes related to PI3K/Akt pathway and levels of phosphorylated Akt in myocardial tissue sample from ischemia area. Results Expressions of many detected genes relating to PI3K/Akt signal pathway were significantly different among the four groups. As compared with group C, 9 genes showed a significant up-regulated expression in group F, 2 genes showed a significant up-regulated expression in group R, and a total number of 33 genes showed a significant up-regulated expression in group F-R. The Western-blot analysis revealed that the expression of phosphorylated Akt in myocardium increased significantly in groups F, R and F-R compared with group C. The expression of phosphorylated Akt in myocardium was also stronger in group F-R than in groups F and R. Conclusions Combined fentanyl postconditioning and RIPOC can produce an enhanced activation of PI3K/Akt signal pathway.