Nrf2信号通路在小鼠内毒素性急性肺损伤时内质网应激反应中的作用  

Role of Nrf2 signaling pathway in endoplasmic reticulum stress response during lipopolysaccharide-induced acute lung injury in mice

作　　者：鲁稳梁[1] 李剑峰[1] 巩红岩[2] 董铁立[1] 艾艳秋[3] Lu Wenliang;Li Jianfeng;Gong Hongyan;Dong Tieli;Ai Yanqiu(Department of Anesthesiology, Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450052, China;Department of Anesthesiology, First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453000,China;Department of Anesthesiology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052,China)

机构地区：[1]郑州大学第二附属医院麻醉科,450014 [2]新乡医学院第一附属医院麻醉科,453100 [3]郑州大学第一附属医院麻醉科,450052

出　　处：《中华麻醉学杂志》2019年第4期494-497,共4页Chinese Journal of Anesthesiology

摘　　要：目的评价核转录因子NF-E2相关因子2(Nrf2)信号通路在小鼠内毒素性急性肺损伤时内质网应激反应中的作用。方法清洁级健康雄性C57BL/6小鼠40只,6~8周龄,体重22~26 g,采用随机数字表法分为4组(n=10):对照组(C组)、急性肺损伤组(ALI组)、salubrinal组(S组)和salubrinal+鸦胆子苦醇组(S+B组)。采用气管内注入LPS 5 mg/kg的方法制备小鼠内毒素性急性肺损伤模型;C组气管内注入生理盐水100 μl;S组于给予LPS后1和24 h时腹腔注射内质网应激反应抑制剂salubrinal 1 mg/kg;S+B组于给予LPS前10 d,隔天腹腔注射鸦胆子苦醇2 mg/kg,余处理同S组。于给予LPS后48 h时处死小鼠取肺,光镜下观察肺组织病理学结果并进行评分,计算肺含水量,采用ELISA法检测IL-17A、TNF-α和IL-6含量,Western blot法检测Nrf2、CCAAT增强子结合蛋白同源蛋白(CHOP)和caspase-12的表达。结果与C组比较,ALI组和S+B组肺含水量、IL-17A、TNF-α和IL-6含量升高,胞浆CHOP和caspase-12表达上调,胞核Nrf2表达下调,S组肺含水量、IL-17A、TNF-α和IL-6含量升高,胞核Nrf2和胞浆CHOP上调,caspase-12表达下调(P<0.05);与ALI组比较,S组肺含水量、IL-17A、TNF-α和IL-6含量降低,胞浆CHOP和caspase-12表达下调,胞核Nrf2表达上调(P<0.05),肺组织病理学损伤减轻;与S组比较,S+B组肺含水量、IL-17A、TNF-α和IL-6含量升高,胞浆CHOP和caspase-12表达上调,胞核Nrf2表达下调(P<0.05),肺组织病理学损伤加重。结论Nrf2信号通路参与了小鼠内毒素性急性肺损伤时内质网应激反应的过程。Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in endoplasmic reticulum stress response during lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Methods Forty clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 22-26 g, were divided into 4 groups (n=10 each) using a random number table method: control group (group C), group ALI, salubrinal group (group S) and salubrinal plus brusatol group (group S+ B). Animals were intratracheally instilled with 5 mg/kg of LPS diluted in normal saline to establish the model of ALI.Animals were intratracheally instilled with 100 μl of normal saline in group C. Mice in group S were intraperitoneally injected with endoplasmic reticulum stress response inhibitor 1 mg/kg salubrinal at 1 and 24 h after LPS instillation.Mice of group S+ B were intraperitoneally injected with brusatol 2 mg/kg once every other day for 10 days prior to LPS instillation, and the other treatments were similar to those previously described in group S. Mice were sacrificed at 48 h after LPS administration, and lungs were removed for microscopic examination of the pathological changes of lung tissues which were scored and for determination of contents of IL-17A, tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in lung tissues (by enzyme-linked immunosorbent assay) and expression of Nrf2, CCAAT/enhancer-binding protein homologous protein (CHOP) and caspase-12 in lung tissues (by Western blot). Lung water content was calculated. Results Compared with group C, the lung water content and contents of IL-17A, TNF-α and IL-6 were significantly increased, the expression of CHOP and caspase-12 in cytoplasma was up-regulated, and the expression of Nrf2 in nuclei was down-regulated in ALI and S+ B groups, and the lung water content and contents of IL-17A, TNF-α and IL-6 were significantly increased, the expression of Nrf2 in nuclei and CHOP in cytoplasma was up-regulated, and the expression of caspase-12 was down-regulated

关 键 词：NF-E2相关因子2 内毒素类 急性肺损伤 内质网应激 

分 类 号：R563.8[医药卫生—呼吸系统]