SIRT1/Nrf2信号通路在小檗碱预处理减轻小鼠肠缺血再灌注损伤中的作用及其与铁死亡的关系  

Role of SIRT1/Nrf2 signaling pathway in berberine preconditioning-induced reduction of intestinal ischemia-reperfusion injury in mice and the relationship with ferroptosis

作　　者：高焕焕 冷玉芳[1,2] 王心润 贾筱晶 Gao Huanhuan;Leng Yufang;Wang Xinrun;Jia Xiaojing(The First Clinical Hospital 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》2023年第12期1515-1519,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82260381);兰州大学医学教育创新发展项目(lzuyxcx-2022-108)。

摘　　要：目的评价沉默信息调节因子1/核因子E2相关因子2(SIRT1/Nrf2)信号通路在小檗碱预处理减轻小鼠肠缺血再灌注损伤中的作用及其与铁死亡的关系。方法SPF级健康雄性C57BL/6小鼠36只,8~10周龄,体质量22~25 g,采用随机数字表法分为6组(n=6):假手术组(S组)、假手术+小檗碱组(SB组)、肠缺血再灌注组(IR组)、小檗碱预处理+肠缺血再灌注组(B组)、小檗碱预处理+SIRT1抑制剂EX527+肠缺血再灌注组(BE组)和小檗碱预处理+铁死亡诱导剂RSL3+肠缺血再灌注组(BR组)。采用夹闭肠系膜上动脉45 min再灌注30 min的方法构建小鼠肠缺血再灌注损伤模型。SB组、B组、BE组和BR组于造模前7 d开始灌胃小檗碱50 mg/kg,1次/d;BE组和BR组于造模前3 d分别腹腔注射EX5275 mg/kg、RSL35 mg/kg,1次/d;其余组予以等量生理盐水。于再灌注30 min时处死小鼠,取小肠组织,光镜下观察肠黏膜病理结果并行Chiu评分,采用比色法检测Fe^(2+)和MDA含量及SOD活性,采用ELISA法检测谷胱甘肽(GSH)含量,采用荧光染色法检测ROS含量,采用Western blot法检测谷胱甘肽过氧化物酶4(GPX4)、SIRT1和Nrf2的表达。结果与S组比较,IR组肠组织Chiu评分升高,Fe^(2+)、MDA和ROS含量升高,GSH含量和SOD活性下降,GPX4表达下调,SIRT1和Nrf2表达上调(P<0.05),SB组Chiu评分差异无统计学意义(P>0.05);与IR组比较,B组肠组织Chiu评分降低,Fe^(2+)、MDA和ROS含量降低,GSH含量和SOD活性升高,GPX4表达上调,SIRT1和Nrf2表达上调(P<0.05);与B组比较,BE组和BR组肠组织Chiu评分升高,Fe^(2+)、MDA和ROS含量升高,GSH含量和SOD活性下降,GPX4表达下调,BE组SIRT1和Nrf2表达下调(P<0.05)。结论小檗碱预处理减轻小鼠肠缺血再灌注损伤的机制可能与激活SIRT1/Nrf2信号通络,进而抑制铁死亡有关。Objective To evaluate the role of silencing information regulatory factor 1/nuclear factor E2 related factor 2(SIRT1/Nrf2)signaling pathway in berberine preconditioning-induced reduction of intestinal ischemia-reperfusion(I/R)injury in mice and the relationship with ferroptosis.Methods Thirty-six SPF-grade healthy male C57BL/6 mice,aged 8-10 weeks,weighing 22-25 g,were divided into 6 groups(n=6 each)by a random number table method:sham operation group(S group),sham operation+berberine preconditioning group(SB group),intestinal I/R group(IR group),intestinal I/R+berberine preconditioning group(B group),intestinal I/R+berberine preconditioning+SIRT1 inhibitor EX527 group(BE group)and berberine preconditioning+intestinal I/R+ferroptosis inducer RSL3 group(BR group).The model of intestinal I/R injury was prepared by clamping the superior mesenteric artery for 45 min followed by 30-min reperfusion in IR group,B group,BE group and BR group,while the superior mesenteric artery was only isolated without ligation in S group and SB group.Berberine 50 mg/kg was administered by intragastric gavage once a day starting from 7 days before developing the model in SB group,B group,BE group and BR group.EX5275 mg/kg and RSL35 mg/kg were intraperitoneally injected once a day at 3 days before surgery in BE group and BR group,respectively.The equal volume of normal saline was given in the other groups.The mice were sacrificed at 30 min of reperfusion,and the intestinal tissues were taken for microscopic examination of the pathological changes of intestinal mucosa(with a light microscope)which was scored according to Chiu and for determination of the contents of Fe^(2+)and malondialdehyde(MDA)and superoxide dismutase(SOD)activity(by colorimetry),glutathione(GSH)content(by enzyme-linked immunosorbent assay),reactive oxygen species(ROS)content(by fluorescence staining),and expression of glutathione peroxidase 4(GPX4),SIRT1 and Nrf2(by Western blot).Results Compared with S group,Chiu′s score was significantly increased,the contents of

关 键 词：小檗碱 肠 再灌注损伤 铁死亡 抗衰老酶1 NF-E2相关因子2 

分 类 号：R965[医药卫生—药理学]