TIPE2在脓毒症小鼠急性肺损伤内源性保护机制中的作用:与TAK1/p38MAPK/NF-κB信号通路的关系  

Role of TIPE2 in endogenous protective mechanism of acute lung injury in septic mice:the relationship with TAK1/p38 MAPK/NF-κB signaling pathway

作　　者：明婷倩 吴晓静[1] 王倩[1] 孔倩[1] 袁敏[1] 陈丽丽 夏中元[1] Ming Tingqian;Wu Xiaojing;Wang Qian;Kong Qian;Yuan Min;Chen Lili;Xia Zhongyuan(Department of Anesthesiology,Renmin Hospital,Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,430060

出　　处：《中华麻醉学杂志》2021年第11期1374-1377,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81970722,81901952);湖北省自然科学基金(2019CFB473)。

摘　　要：目的评价肿瘤坏死因子α诱导蛋白家族8-样蛋白2(TIPE2)在脓毒症小鼠急性肺损伤内源性保护机制中的作用及其与转化生长因子-β激活的蛋白激酶(TAK1)/p38-丝裂原活化的磷酸激酶(p38 MAPK)/NF-κB信号通路的关系。方法雄性野生型小鼠和TIPE2基因敲除小鼠各16只,6~8周龄,体重20~25 g,采用随机数字表法分别分为野生型假手术组(WT-sham组)、野生型ALI组(WT-ALI组)、TIPE2基因敲除型假手术组(KO-sham组)和TIPE2基因敲除急性肺损伤组(KO-ALI组),每组8只。采用盲肠结扎穿孔法制备脓毒症小鼠急性肺损伤模型。于术后24 h时处死小鼠并留取左侧颈总动脉血标本和肺组织标本。HE染色观察肺组织病理结果并进行肺损伤评分;采集左侧颈总动脉血进行血气分析并计算氧合指数(OI);检测每组小鼠肺泡灌洗液中性粒细胞(PMN)计数;采用Western blot法检测肺组织TIPE2、p-TAK1、p-p38 MAPK和p-NF-κB p65的表达;ELISA法检测血清IL-1β和IL-6的浓度。结果与WT-sham组比较,WT-ALI组和KO-ALI组小鼠肺损伤评分、PMN计数、肺组织p-TAK1、p-p38 MAPK和p-NF-κB p65表达水平和血清IL-1β和IL-6浓度升高,PaO_(2)、OI和肺组织TIPE2表达水平降低(P<0.05);与WT-ALI组比较,KO-ALI组小鼠肺损伤评分、PMN计数、肺组织p-TAK1、p-p38 MAPK和p-NF-κB p65表达水平和血清IL-1β和IL-6浓度升高,PaO_(2)、OI和肺组织TIPE2表达水平降低(P<0.05)。结论TIPE2参与了脓毒症小鼠急性肺损伤的内源性保护机制,与抑制TAK1/p38 MAPK/NF-κB信号通路激活有关。Objective To evaluate the role of tumor necrosis factor-alpha-induced protein-8 like-2(TIPE2)in endogenous protective mechanism of acute lung injury(ALI)in septic mice and the relationship with transforming growth factor-β-activated protein kinase(TAK1)/p38-mitogen-activated phosphokinase(p38 MAPK)/nuclear factorκB(NF-κB)signaling pathway.Methods Sixteen male wild-type mice and 16 TIPE2-knockout mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 4 groups:wild-type sham operation group(group WT-sham),wild-type ALI group(group WT-ALI),TIPE2-knockout sham operation group(group KO-sham)and TIPE2-knockout ALI group(group KO-ALI),with 8 mice in each group.The ALI model was established by cecal ligation and perforation(CLP)in septic mice.Mice were sacrificed at 24 h after CLP to acquire left carotid artery blood samples and lung tissue sections.Lung tissue sections were stained with hematoxylin&eosin(H&E)to examine the pathological changes and lung injury scores were assessed.Arterial blood samples were collected from the left carotid artery for blood gas analysis,and oxygenation index(OI)was calculated.Bronchoalveolar lavage fluid was collected to measure polymorphonuclear neutrophil(PMN)count.The expression of TIPE2,phosphorylated TAK1(p-TAK1),phosphorylated p38 MAPK(p-p38)MAPK and phosphorylated NF-κB p65(p-NF-κB p65)was detected using Western blot analysis.The levels of interleukin-1beta(IL-1β)and IL-6 in serum were determined by enzyme-linked immunosorbent assay.Results Compared with group WT-sham,the lung injury score,PMN counts and concentrations of IL-1βand IL-6 in serum were significantly increased,the expression of p-TAK1,p-p38 MAPK and p-NF-κB p65 was up-regulated,the PaO_(2) and OI were decreased,and the expression of TIPE2 was down-regulated in WT-ALI and KO-ALI groups(P<0.05).Compared with group WT-ALI,the lung injury score,PMN counts and concentrations of IL-1βand IL-6 in serum were significantly increased,the expression of p-TAK1,p-p38 MAPK and p-NF-κB p65 was up-regulated,the PaO_

关 键 词：肿瘤坏死因子α诱导蛋白家族8-样蛋白2 脓毒症 急性肺损伤 转化生长因子β 蛋白激酶类 P38丝裂原活化蛋白激酶类 NF-κB 

分 类 号：R459.7[医药卫生—急诊医学]