组蛋白去乙酰化酶在丁酸钠减轻小鼠肠缺血再灌注损伤中的作用:与氧化应激和细胞凋亡的关系

Role of histone deacetylase in sodium butyrate-induced reduction of intestinal ischemia-reperfusion injury in mice:relationship with oxidative stress and cell apoptosis

目的评价组蛋白去乙酰化酶(HDAC)在丁酸钠减轻小鼠肠缺血再灌注损伤中的作用及其与氧化应激和细胞凋亡的关系。方法SPF级健康雄性C57BL小鼠24只,6~8周龄,体重22~25 g,根据随机数字表法分成4组(n=6):假手术组(S组)、肠缺血再灌注组(IR组)、肠缺血再灌注+丁酸钠组(IN组)和肠缺血再灌注+ITSA-1+丁酸钠组(INI组)。IR组、IN组和INI组采用夹闭肠系膜上动脉45 min再灌注2 h的方法制备小鼠肠缺血再灌注损伤模型,S组仅分离肠系膜上动脉,不结扎。于模型制备前1周开始IN组和INI组小鼠给予500 mg/kg丁酸钠灌胃,1次/d,INI组腹腔注射HDAC激动剂ITSA-10.5 mg/kg,3次/周,其余组给予等量生理盐水。于再灌注2 h时处死小鼠取小肠组织,HE染色光学显微镜下观察病理学结果,并进行Chiu评分,采用ELISA法测定MDA含量,采用Western blot法检测cleaved caspase-3表达。结果与S组比较,IR组、IN组和INI组Chiu评分升高,小肠组织cleaved caspase-3表达上调,IR组和INI组小肠组织MDA含量升高(P<0.05);与IR组比较,IN组和INI组Chiu评分降低,IN组小肠组织MDA含量降低,cleaved caspase-3表达下调(P<0.05);与IN组比较,INI组Chiu评分升高,小肠组织MDA含量升高,cleaved caspase-3表达上调(P<0.05)。结论HDAC参与了丁酸钠减轻小鼠肠缺血再灌注损伤的过程,与抑制氧化应激和细胞凋亡有关。

Objective To evaluate the role of histone deacetylase(HDAC)in sodium butyrate-induced reduction of intestinal ischemia-reperfusion(I/R)injury in mice and the relationship with oxidative stress and cell apoptosis.Methods Twenty-four SPF healthy male C57BL mice,aged 6-8 weeks,weighing 22-25 g,were divided into 4 groups(n=6 each)according to the random number table method:sham operation group(S group),intestinal I/R group(IR group),intestinal I/R+sodium butyrate group(IN group)and intestinal I/R+ITSA-1+sodium butyrate group(INI group).In IR,IN and INI groups,the superior mesenteric artery was clamped for 45 min,followed by reperfusion for 2 h to prepare the model of intestinal I/R injury,while the superior mesenteric artery was only isolated without ligation in S group.One week before preparation of the model,sodium butyrate 500 mg/kg was intragastrically administered once a day in IN group and INI group,the HDAC activator ITSA-10.5 mg/kg was intraperitoneally injected three times a week in INI group,and the equal volume of normal saline was given instead in the other groups.The mice were sacrificed at 2 h of reperfusion and small intestinal tissues were obtained for microscopic examination of the pathological changes which were assessed using Chiu′s score and for determination of the content of MDA(by enzyme-linked immunosorbent assay)and expression of cleaved caspase-3(by Western blot).Results Compared with S group,Chiu′s score was significantly increased,and the expression of cleaved caspase-3 was up-regulated in IR,IN and INI groups,the content of MDA in small intestinal tissues was significantly increased in IR and INI groups(P<0.05).Compared with IR group,Chiu′s score was significantly decreased in IN and INI groups,and the content of MDA was significantly decreased,and the expression of cleaved caspase-3 was down-regulated in IN group(P<0.05).Compared with IN group,Chiu′s score and content of MDA were significantly increased,and the expression of cleaved caspase-3 was up-regulated in INI group(P<0.05).Conclusions HDAC is involved in sodium butyrate-induced reduction of intestinal I/R injury in mice,which is related to the inhibition of oxidative stress and cell apoptosis.