HO-1/CO信号通路在大鼠内毒素性急性肺损伤时内质网应激中的作用  

Role of endogenous heme oxygenase-1/carbon monoxide signaling pathway in endoplasmic reticulum stress during endotoxin-induced acute lung injury in rats

作　　者：宫丽荣[1] 吴丽丽[1] 穆蕊 宗春辉[1] 杜诗涵 张艳芳[1] 余剑波[1] Gong Lirong;Wu Lili;Mu Rui;Zong Chunhui;Du Shihan;Zhang Yanfang;Yu Jianbo(Department of Anesthesiology,Tianjin Nankai Hospital Nankai Clinical College of Tianjin Medical University,Tianjin 300100,China)

机构地区：[1]天津医科大学南开临床学院天津市南开医院麻醉科,300100

出　　处：《中华麻醉学杂志》2019年第6期734-737,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81601707,81772106).

摘　　要：目的评价内源性血红素氧合酶-1/一氧化碳(HO-1/CO)信号通路在大鼠内毒素性急性肺损伤(ALI)时内质网应激中的作用.方法健康清洁级雄性SD大鼠40只,体重190~210 g,8周龄,采用随机数字表法分为4组(n=10):对照组(C组)、ALI组、ALI+锌原卟啉-IX组(AZ组)和ALI+溶剂碳酸氢钠组(AV组).采用静脉注射LPS 5 mg/kg的方法制备内毒素性ALI模型.模型制备前30 min时AZ组腹腔注射锌原卟啉-IX10 mol/kg(50 mmol/L碳酸氢钠溶液稀释至1 ml),AV组腹腔注射50 mmol/L碳酸氢钠溶液1 ml.注射LPS 6 h后取颈总动脉血样并处死大鼠取肺组织,测定血浆和肺组织CO水平,光镜下观察病理学结果,并进行肺损伤评分,确定湿重/干重(W/D)比值,采用TUNEL法测定细胞凋亡情况,计算细胞凋亡指数(AI),采用Western blot法检测肺组织HO-1、葡萄糖调节蛋白78(GRP78)、p-PERK、p-elF2a、CCAAT/增强子结合蛋白同源蛋白(CHOP)及caspase-12的表达水平.结果与C组比较,其余3组肺损伤评分、肺组织W/D比值及AI升高,血浆及肺组织CO水平升高,肺组织HO-1、GRP78、p-PERK、p-elF2a、CHOP及caspase-12表达上调(P<0.05);与ALI组比较,AZ组肺损伤评分、W/D比值及AI升高,血浆及肺组织CO水平降低,肺组织HO-1表达下调,GRP78、p-PERK、p-elF2a、CHOP及caspase-12表达上调(P<0.05),AV组上述各指标差异无统计学意义(P>0.05).结论 HO-1/CO信号通路可能通过抑制内质网应激,对内毒素性ALI大鼠发挥内源性保护作用.Objective To evaluate the role of endogenous heme oxygenase-1/carbon monoxide ( HO-1/CO) signaling pathway in endoplasmic reticulum stress during endotoxin-induced acute lung injury ( ALI) in rats. Methods Forty healthy clean-grade male Sprague-Dawley rats, aged 8 weeks, weighing 190-210 g, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), ALI group, ALI plus ZnPP-IX group (group AZ), and ALI plus vehicle sodium bicarbonate group ( group AV). ALI was induced by intravenously injecting lipopolysaccharide 5 mg/kg in anesthetized rats. At 30 min before establishing the model, ZnPP-IX 10μmol/kg (diluted to 1 ml in 50 mmol/L sodium bicarbonate) was intraperitoneally injected in group AZ, and 50 mmol/L sodium bicarbonate 1 ml was intra-peritoneally injected in group AV. After injecting lipopolysaccharide for 6 h, blood samples were collected from the common carotid artery for determination of plasma CO concentration, the rats were then sacrificed, and lungs were removed for microscopic examination of the pathological changes which were scored and for determination of CO level, wet to dry weight ratio ( W/D ratio), cell apoptosis ( by TUNEL), and expres-sion of heme oxygenase-1 ( HO-1), glucose-regulated protein 78 ( GRP78), phosphorylated protein kinase R-like endoplasmie reticulum kinase (p-PERK), phosphorylated eukaryotic translation initiation factor 2 alpha ( p-elF2 ), CCAAT/enhancer-binding protein homologous protein ( CHOP ) and caspase-12 in lung tissues ( by Western blot). Apoptosis index ( AI) was calculated. Results Compared with group C, the lung injury scores, W/D ratio, AI and CO levels in plasma and lung tissues were significantly increased, and the expression of HO-1, GRP78, p-PERK, p-elF2, CHOP and caspase-12 was up-regulated in the other three groups ( P<0. 05). Compared with group ALI, lung injury scores, W/D ratio and AI were sig-nificantly increased, CO levels in plasma and lung tissues were decreased, the expression of HO-1 was down-regulated, and

关 键 词：血红素加氧酶-1 一氧化碳 内毒素类 急性肺损伤 内质网应激 

分 类 号：R563[医药卫生—呼吸系统]