酚妥拉明对肠缺血再灌注损伤小鼠肠黏膜屏障及组织灌注的影响

Effect of phentolamine on gut barrier and perfusion in mice after intestinal ischemia-reperfusion injury

目的:探讨酚妥拉明(Regitine)对肠缺血再灌注(Ⅱ/R)损伤小鼠肠黏膜屏障和组织灌注的保护作用。方法:采用夹闭肠系膜上动脉方法建立Ⅱ/R模型,将32只小鼠随机分为四组:对照组、Ⅱ/R组(肠缺血再灌注)、Regitine组(Ⅱ/R+酚妥拉明肠系膜根部注射)和Regitineⅳ组(Ⅱ/R+酚妥拉明尾静脉注射),每组8只。观察各组肠道炎症因子(IL-6、TNF-α)、血清肠损伤标记物水平(D-lactate、I-FABP),肠道血流灌注指数、平均动脉压(MAP)、血清血管内皮损伤标记物(Syndecan-1),组织病理评分、肠黏膜屏障通透性(FD4)、紧密连接蛋白(Claudin-1、Occludin)表达变化,肠组织细胞凋亡(TUNEL)、线粒体自噬水平(p62、mito-LC3、Tim23)以及细胞色素C(Cyt-c)泄露水平。结果:与Ⅱ/R组相比,Regitine组和Regitineⅳ组小鼠肠道IL-6、TNF-α、血清D-lactate、I-FABP、Syndecan-1、FD4、组织病理评分、肠组织TUNEL以及Cyt-c泄露减少(P<0.05),肠道血流灌注指数、紧密连接蛋白表达、线粒体自噬水平升高,差别具有统计学意义(P<0.05);与Regitineⅳ组相比,Regitine组肠道IL-6、血清D-lactate、I-FABP、FD4水平、肠组织TUNEL及Cyt-c泄露水平降低(P<0.05),同时肠道血流灌注指数、MAP、Occludin表达、线粒体自噬水平升高(P<0.05)。结论:酚妥拉明对Ⅱ/R小鼠肠黏膜屏障和组织灌注具有一定的保护作用,肠系膜根部局部给药效果优于尾静脉全身给药,该发现有望为保护Ⅱ/R后肠黏膜屏障功能和组织灌注提供新的治疗思路。

Objective:To investigate the protective effect of phentolamine on gut barrier and tissue perfusion in mice after intestinal ischemia-reperfusion(II/R)injury.Methods:The II/R model was established by clipping the superior mesenteric artery.32 mice were randomly divided into four groups:control group,II/R group(intestinal ischemia-reperfusion),Regitine group(II/R+phentolamine,root of mesentery injection)and Regitine iv group(II/R+phentolamine,tail vein injection).Intestinal inflammatory factors(IL-6,TNF-α),serum intestinal injury markers(D-lactate,I-FABP),intestinal blood perfusion index,mean arterial pressure(MAP),serum vascular endothelial injury marker(Syndecan-1),histopathological score,gut barrier permeability(FD4),expression changes of tight junction proteins(Claudin-1,Occludin),intestinal tissue apoptosis(TUNEL),mitophagy levels(p62,mito-LC3,Tim23)and cytochrome C(Cyt-c)leakage levels were studied.Results:Compared with II/R group,the intestinal IL-6,TNF-α,serum D-lactate,I-FABP,Syndecan-1,FD4,histopathological score,TUNEL and Cyt-c leakage decreased in the Regitine group and the Regitine iv group(P<0.05).Intestinal blood perfusion index,tight junction protein expression,and mitophagy levels increased in the Regitine group and the Regitine iv group compare with the II/R group(P<0.05).Intestinal IL-6,serum D-lactate,I-FABP,FD4 level,TUNEL and Cyt-c leakage levels decreased in the Regitine group compare with the Regitine iv group(P<0.05).In addition,the intestinal blood perfusion index,MAP,Occludin expression,mitophagy level were increased in Regitine group(P<0.05).Conclusion:Phentolamine has a certain protective effect on the gut barrier and tissue perfusion in II/R mice.The effect of local administration(mesentery root injection)is better than systemic administration.This finding may provide new therapeutic options for protecting gut barrier function and tissue perfusion after II/R injury.