组蛋白去乙酰化酶6在丁酸钠减轻小鼠肠缺血再灌注损伤中的作用  

Role of histone deacetylase 6 in reduction of intestinal ischemia-reperfusion injury by sodium butyrate in mice

作　　者：吕兴娇 冷玉芳[1,2] 韩晓霞 侯小煜 曹雪芬 NIBARUTA JANVIER Lyu Xingjiao;Leng Yufang;Han Xiaoxia;Hou Xiaoyu;Cao Xuefen;Janvier NIBARUTA(The First Clinical Medical College of Lanzhou University,Lanzhou 730000,China;Department of Anesthesiology,The First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一临床医学院,兰州730000 [2]兰州大学第一医院麻醉科,兰州730000

出　　处：《中华麻醉学杂志》2022年第6期690-694,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81960345);甘肃省自然科学基金(21JR1RA069);兰州大学第一医院院内基金(ldyyyn2019-25)。

摘　　要：目的评价组蛋白去乙酰化酶6(HDAC6)在丁酸钠减轻小鼠肠缺血再灌注损伤中的作用。方法SPF级健康成年雄性C57BL/6小鼠24只,8~10周龄,体重22~25 g,采用随机数字表法分为4组(n=6):假手术组(S组)、肠缺血再灌注组(I/R组)、肠缺血再灌注+丁酸钠组(I/R+SB组)和肠缺血再灌注+ITSA-1+丁酸钠组(I/R+I+SB组)。采用夹闭肠系膜上动脉45 min、再灌注120 min的方法建立肠缺血再灌注损伤模型。I/R+I+SB组分别于缺血前6、3和1 d时腹腔注射HDACs激活剂ITSA-10.5 mg/kg;I/R+SB组和I/R+I+SB组缺血前1周每天灌胃给予丁酸钠500 mg/kg,S组和I/R组予以等量生理盐水。再灌注120 min时心脏取血后处死小鼠,取小肠组织,采用ELISA法检测血清和小肠组织二胺氧化酶(DAO)水平;光镜下观察小肠组织病理学结果并进行Chiu′s评分;采用Western blot法测定小肠组织微管相关蛋白轻链3Ⅱ(LC3Ⅱ)、P62和HDAC6的表达水平;采用ELISA法检测小肠组织组蛋白H3(H3)和乙酰化组蛋白H3(Ac-H3)的含量。结果与S组比较,I/R组小肠组织Chiu′s评分、血清和小肠组织DAO水平升高,小肠组织LC3Ⅱ和HDAC6表达上调,P62表达下调,H3含量升高,Ac-H3含量降低(P<0.05);与I/R组比较,I/R+SB组小肠组织Chiu′s评分、血清和小肠组织DAO水平降低,小肠组织LC3Ⅱ和HDAC6表达下调,P62表达上调,H3含量降低,Ac-H3含量升高(P<0.05);与I/R+SB组比较,I/R+I+SB组小肠组织Chiu′s评分、血清和小肠组织DAO水平升高,小肠组织LC3Ⅱ和HDAC6表达上调,P62表达下调,H3含量升高,Ac-H3含量降低(P<0.05)。结论丁酸钠可通过抑制HDAC6活性减轻小鼠肠缺血再灌注损伤,其机制可能与抑制自噬和促进H3乙酰化有关。Objective To evaluate the role of histone deacetylase 6(HDAC6)in reduction of intestinal ischemia-reperfusion(I/R)injury by sodium butyrate in mice.Methods Twenty-four SPF healthy adult male C57BL/6 mice,aged 8-10 weeks,weighing 22-25 g,were divided into 4 groups(n=6 each)by the random number table method:sham operation group(S group),intestinal I/R group(I/R group),intestinal I/R+sodium butyrate group(I/R+SB group),and intestinal I/R+ITSA-1+sodium butyrate group(I/R+I+SB group).The model of intestinal I/R injury was established by clipping superior mesenteric artery for 45 min followed by 120 min of reperfusion in anesthetized animals.In I/R+I+SB group,the HDACs activator ITSA-10.5 mg/kg was intraperitoneally injected at 6,3 and 1 days before ischemia.Sodium butyrate 500 mg/kg was given by intragastric administration every day one week before ischemia in I/R+SB group and I/R+I+SB group,and the equal volume of normal saline was given in S group and I/R group.At 120 min of reperfusion,the mice were sacrificed and their small intestine tissues were obtained.The levels of diamine oxidase(DAO)in serum and intestinal tissues were detected by enzyme-linked immunosorbent assay.The pathological changes of small intestinal tissues were observed with a light microscope,and intestinal damage was assessed and scored according to Chiu.The expression of microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ),P62 and HDAC6 was determined by Western blot.The contents of histone H3(H3)and acetylated histone H3(Ac-H3)in small intestinal tissues were determined by enzyme-linked immunosorbent assay.Results Compared with S group,the Chiu′s score,levels of DAO in serum and small intestinal tissues were significantly increased,the expression of LC3Ⅱand HDAC6 was up-regulated,P62 expression was down-regulated,H3 content was increased,and AC-H3 content was decreased in I/R group(P<0.05).Compared with I/R group,the Chiu′s score,levels of DAO in serum and small intestinal tissues were significantly decreased,the expression of LC3�

关 键 词：丁酸盐类 再灌注损伤 肠 组蛋白脱乙酰基酶类 

分 类 号：R574[医药卫生—消化系统]