叉头状转录因子O1/p38丝裂原活化蛋白激酶信号通路在脂多糖致急性肺损伤中的作用  

Role of the forkhead transcription factor O1/p38 mitogen activated protein kinase signaling pathway in acute lung injury induced by lipopolysaccharide

作　　者：刘斌[1] 徐丽丽[2] 张红[3] 欧阳军[4] 杨娟娟 马猛 王全明 白彦芳 邬超[5] Liu Bin;Xu Lili;Zhang Hong;Ouyang Jun;Yang Juanjuan;Ma Meng;Wang Quanming;Bai Yanfang;Wu Chao(Department of Emergency surgery,Zhu Xianyi Memorial Hospital(Metabolic Disease Hospital)of Tianjin Medical University,Tianjin 300134,China;Department of General Medicine,First Affiliated Hospital of Shihezi University,Shihezi 830002,China;Department of Medical,the First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China;Emergency Medical Center,First Affiliated Hospital of Shihezi University,Shihezi 830002,China;Department of Respiratory and Critical Care Medicine,First Affiliated Hospital of Shihezi University,Shihezi 830002,China)

机构地区：[1]天津医科大学朱宪彝纪念医院(代谢病医院)急诊外科,天津300134 [2]石河子大学第一附属医院全科医学科,石河子830002 [3]成都医学院第一附属医院医务部,成都610500 [4]石河子大学第一附属医院急诊医学中心,石河子830002 [5]石河子大学第一附属医院呼吸与危重症医学科,石河子830002

出　　处：《中华实验外科杂志》2024年第10期2265-2268,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81960005.8196038781760017)。

摘　　要：目的:探讨叉头状转录因子O1(FoxO1)/p38丝裂原活化蛋白激酶(p38 MAPK)信号通路在脂多糖致急性肺损伤中的作用。方法:按照随机数字表法将20只雄性无特定病原体(SPF)级C57BL/6J小鼠分为正常组及模型组,每组10只,模型组采用脂多糖气管滴注小鼠复制急性肺损伤模型,正常组给予等量生理盐水。分类计数小鼠支气管肺泡灌洗液(BALF)中的细胞,肺损伤评分及肺湿/干重比,苏木精-伊红(HE)染色检测肺组织病理形态,酶联免疫吸附实验(ELISA)检测肺组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6含量,免疫印迹法检测FoxO1及pp38 MAPK表达。将小干扰RNA(siRNA)NC和siRNA FoxO1转染至BEAS-2B人正常肺上皮细胞中,实时荧光定量聚合酶链反应(RT-qPCR)检测FoxO1 mRNA表达,免疫印迹法检测FoxO1及pp38 MAPK表达,ELISA检测细胞中TNF-α、IL-1β、IL-6含量。组间比较采用t检验。结果:正常组小鼠肺泡完整;模型组小鼠肺泡断裂,大量炎性细胞浸润,肺泡间隙增宽并充血水肿严重。模型组病理评分(13.02±1.09比0.64±0.02,t=6.316,P<0.01)、湿重/干重比(18.52±1.85比1.89±0.19,t=5.987,P<0.01)、总蛋白含量[(252.46±25.25)g/L比(45.28±4.53)g/L,t=6.326,P<0.01]、中性粒细胞[(8.35±0.83)×10^(8)/L比(2.73±0.28)×10^(8)/L,t=5.326,P<0.01]、白细胞总数[(13.29±1.33)×10^(8)/L比(4.50±0.45)×10^(8)/L,t=6.127,P<0.01]、TNF-α[(14.24±1.38)μg/L比(1.41±0.14)μg/L,t=6.124,P<0.01]、IL-1β[(280.31±28.03)pg/ml比(25.34±2.53)pg/ml,t=5.684,P<0.01]、IL-6[(263.35±20.63)ng/ml比(73.41±7.34)ng/ml,t=5.654,P<0.01]含量、FoxO1(0.41±0.02比0.12±0.01,t=6.421,P<0.01)及pp38 MAPK(0.89±0.03比0.21±0.01,t=5.864,P<0.01)蛋白表达量高于正常组。干扰组FoxO1(0.35±0.02比1.00±0.07,t=6.851,P<0.01)mRNA表达量、FoxO1(0.46±0.10比0.98±0.05,t=6.147,P<0.01)、pp38 MAPK(1.87±0.01比0.97±0.06,t=5.840,P<0.01)蛋白表达量、TNF-α[(1.23±0.11)μg/L比(8.35±0.45)μg/L,t=5.987,P<0.01]、IL-1β[(2.65±0Objective:To role of the forkhead transcription factor O1(FoxO1)/p38 mitogen activated protein kinase(p38 MAPK)signaling pathway in acute lung injury induced by lipopolysaccharide(LPS).Methods:Twenty male specific pathogen free(SPF)grade C57BL/6J mice were divided into a normal group and a model group,10 per group.The model group received intratracheal instillation lipopolysaccharide to replicate an acute lung injury model.The normal group was given an equal amount of physiological saline.The cells in bronchoalveolar lavage fluid(BALF),lung injury score,and lung wet/dry weight ratio was detected.The pathological morphology of lung tissue was detected by hematoxylin and eosin(HE)staining.The level of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-1β,IL-6 in lung tissue was detected by enzyme-linked immunosorbent assay(ELISA).The expression of FoxO1 and pp38 MAPK was detected by Western blotting.Transfection small interfering RNA(siRNA)NC and siRNA FoxO1 into BEAS-2B human normal lung epithelial cells,the expression of FoxO1 mRNA was detected by real-time fluorescent quantitative polymerase chain reaction(RT-qPCR).The expression of FoxO1 and pp38 MAPK was detected by western blot.The level of TNF-α,IL-1β,IL-6 was detected yb ELISA.Group t test was used to analyze differences between groups.Results:Mices had intact alveoli in normal group.Mices in model group had alveolar rupture,extensive infiltration of inflammatory cells,widened alveolar spaces,and severe congestion and edema.Pathological score(13.02±1.09比0.64±0.02,t=6.316,P<0.01),W/D(18.52±1.85 vs.1.89±0.19,t=5.987,P<0.01),the total protein content[(252.46±25.25)g/L vs.(45.28±4.53)g/L,t=6.326,P<0.01],neutrophils number[(8.35±0.83)×10^(8)/L vs.(2.73±0.28)×10^(8)/L,t=5.326,P<0.01],the total leukocyte number[(13.29±1.33)×10^(8)/L vs.(4.50±0.45)×10^(8)/L,t=6.127,P<0.01],the level of TNF-α[(14.24±1.38)μg/L vs.(1.41±0.14)μg/L,t=6.124,P<0.01],IL-1β[(280.31±28.03)pg/ml vs.(25.34±2.53)pg/ml,t=5.684,P<0.01],IL-6[(263.35±20.63)ng/ml vs.(73.

关 键 词：叉头状转录因子O1 P38丝裂原活化蛋白激酶 急性肺损伤 炎症 

分 类 号：R563[医药卫生—呼吸系统]