Nrf2／HO-1信号通路在远端缺血预处理减轻小鼠内毒素性急性肺损伤中的作用  

Role of Nrf2/HO-1 signaling pathway in remote ischemic preconditioning-induced reduction of li-popolysaccharide-induced acute lung injury in mice

作　　者：巩红岩[1] 郑芳[2] 贾志杰 刘景景[1] 董铁立[4] 赵国安[3] Gong Hongyan;Zheng Fang;Jia Zhijie;Liu Jingjing;Dong Tieli;Zhao Guoan(Department of Anesthesiology, First Affiliated Hospital oJ X~nxtang Meatcat untvers~y, A~nx~ang .~J.~t~t~L,, Chin;Department of Ultrasound, First Affiliated Hospital of Xinxiang Medi- cal University, Xinxiang 453000, Chin;Department of Coronary Heart Disease Center, First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453000, China;Department of Anesthesiology, Second Affiliated Hospital of Zhengzhou, Zhengzhou 450003, China)

机构地区：[1]新乡医学院第一附属医院麻醉科,453000 [2]新乡医学院第一附属医院超声科,453000 [3]新乡医学院第一附属医院心血管研究中心,453000 [4]郑州大学第二附属医院麻醉科,450003

出　　处：《中华麻醉学杂志》2018年第2期245-249,共5页Chinese Journal of Anesthesiology

摘　　要：目的 评价核转录因子NF-E2相关因子2/血红素加氧酶-1（Nrf2/HO-1）信号通路在远端缺血预处理减轻小鼠内毒素性急性肺损伤中的作用.方法 健康雄性C57BL/6小鼠68只,6-8周龄,体重22- 26 g,采取随机数字表法分为4组（n=17）：对照组（C组）、急性肺损伤组（ALI组）、远端缺血预处理组（RIPC组）、鸦胆子苦醇＋远端缺血预处理组（B＋RIPC组）.C组气管内注入生理盐水100μl;ALI组气管内注入LPS 5 mg/kg制备急性肺损伤模型;RIPC组于制备急性肺损伤模型前1h,施行远端缺血预处理：将止血带置于小鼠右后肢,充气5 min后放气5 min,共6个循环;B组于制备急性肺损伤模型前10 d开始,隔天腹腔注射Nrf2抑制剂鸦胆子苦醇2 mg/kg（溶于1％二甲基亚砜100μl中）;B＋RIPC组于制备急性肺损伤模型前10d,隔天腹腔注射鸦胆子苦醇2 mg/kg,模型制备前1h时行远端缺血预处理.模型制备后24 h时,收集支气管肺泡灌洗液（BALF）,测定蛋白浓度,并行中性粒细胞计数;然后处死小鼠,取肺组织,计算含水率,光镜下观察病理学结果,采用比色法测定MPO活性,采用ELISA法检测IL-1β和TNF-α的含量,采用Western blot法测定Nrf2、HO-1和高迁移率族蛋白B1（HMGB1）的表达水平.结果 与C组比较,ALI组肺组织含水率、MPO活性、IL-1β和TNF-α含量、BALF中性粒细胞计数和蛋白浓度升高,肺组织Nrf2、HO-1和HMGB1表达上调（P＜0.05）;与ALI组比较,RIPC组肺组织含水率、MPO活性、IL-1β和TNF-α含量、BALF中性粒细胞计数和蛋白浓度降低,肺组织Nrf2和HO-1表达上调,HMGB1表达下调（P＜0.05）,病理学损伤减轻;与RIPC组比较,B＋RIPC组组肺组织含水率、MPO活性、IL-1β和TNF-α含量、BALF中性粒细胞计数和蛋白浓度升高,肺组织Nrf2和HO-1表达下调,HMGB1表达上调（P＜0.05）,病理学损伤加重.结论Nrf2/HO-1信号通路的激活参与了远端缺血预处理减轻大鼠内毒素性急性肺损伤.Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway in remote ischemic preconditioning-induced reduction of lipopolysaccharide （LPS）-induced acute lung injury （ALI） in mice.Methods Sixty-eight healthy male C57BL/6 mice,aged 6-8 weeks,weighing 22-26 g,were divided into 4 groups （n =17 each） using a random number table：control group （group C）,ALI group,remote ischemic preconditioning group （group RIPC） and brusatol plus remote ischemic preconditioning group （group B＋RIPC）.Normal saline 100 μl was intratracheally instilled in group C.ALI was induced by intratracheal instillation of LPS 5 mg/kg in group ALI.Mice in group RIPC were subjected to 6 cycles of 5-min ischemia followed by 5-min reperfusion in the right hindlimbs using a tourniquet,and 1 h later the model of ALI was established.Nrf2 inhibitor brusatol 2 mg/kg （in 100 μl of 1% dimethyl sulfoxide） was intraperitoneally injected every other day for 10 days prior to establishment of the ALI model in group B.Brusatol 2 mg/kg was intraperitoneally injected every other day for 10 days prior to establishment of the ALI model,and remote ischemic preconditioning was performed at 1 h before establishment of the ALI model in group B＋RIPC.Seven mice in each group were selected at 24 h after establishment of the ALI model,and bronchoalveolar lavage fluid （BALF） was collected for determination of protein concentrations and neutrophil count.Mice were then sacrificed and lungs were removed for determination of lung water content,myeloperoxidase （MPO） activity,contents of interleukin-1beta （IL-1β） and tumor necrosis factor-alpha （TNF-α）,and expression of Nrf2,HO-1 and high-mobility group box 1 protein （HMGB1） in lung tissues （by Western blot） and for examination of pathological changes （with a light microscope）.Results Compared with group C,the lung water content,MPO activity,contents of IL-1β and TNF-α,and neutrophil count and

关 键 词：NF-E2相关因子2 血红素加氧酶-1 缺血预处理 呼吸窘迫综合征 成人 内毒素类 

分 类 号：R614[医药卫生—麻醉学]