血红素加氧酶⁃1通过调控bHLH亮氨酸拉链转录因子E3/高尔基体应激反应减轻小鼠内毒素性急性肺损伤  

Heme oxygenase‑1 attenuates endotoxin‑induced acute lung injury through regulating bHLH leucine zipper transcription factor E3/Golgi stress response in mice

作　　者：张野 李香云 黄炎 吴丽丽[1] 武丽娜[1] 余剑波[1] Zhang Ye;Li Xiangyun;Huang Yan;Wu Lili;Wu Lina;Yu Jianbo(Department of Anesthesiology and Critical Care Medicine,Naikai Clinical College of Tianjin Medical University(Tianjin Nankai Hospi-tal),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)麻醉与重症医学科,天津300100

出　　处：《国际麻醉学与复苏杂志》2024年第1期11-17,共7页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金青年项目(82102247);第八届中国科协青年人才托举工程项目(2022QNRC001)。

摘　　要：目的探讨在小鼠内毒素性急性肺损伤(ALI)中血红素加氧酶⁃1(HO⁃1)对bHLH亮氨酸拉链转录因子E3(TFE3)表达与核转位以及高尔基体应激反应的影响。方法将24只雄性C57BL/6小鼠按随机数字表法分为4组(每组6只):空白对照组(Ctrl组)、内毒素[脂多糖(lipopolysaccharide,LPS)]急性肺损伤组(LPS组)、内毒素性急性肺损伤+HO⁃1激动剂氯高铁血红素(Hemin)组(LPS+Hemin组)和Hemin组。Ctrl组尾静脉注射生理盐水0.5 ml;LPS组尾静脉注射LPS 10 mg/kg建立小鼠内毒素性ALI模型;LPS+Hemin组腹腔注射Hemin 50 mg/kg,1 h后尾静脉注射LPS 10 mg/kg建立内毒素性ALI模型;Hemin组腹腔注射Hemin 50 mg/kg。造模12 h后对小鼠进行断颈处死,收取肺组织。苏木精⁃伊红染色(H⁃E染色)观察小鼠肺组织病理学变化并行肺损伤评分;计算肺组织湿重/干重(W/D)值;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测肺组织细胞凋亡指数;酶联免疫吸附测定(ELISA)法检测肺组织白细胞介素(IL)⁃6、肿瘤坏死因子(TNF)⁃α含量;流式细胞仪检测肺组织活性氧(ROS)的含量;免疫荧光染色观察TFE3核转位情况;蛋白质免疫印迹法(Western blot)测定HO⁃1、TFE3、高尔基体基质蛋白130(GM130)、高尔基体重组和堆叠蛋白65(GRASP65)、囊泡转运蛋白小GTP酶20(RAB20)、突触融合蛋白3(STX3A)、WD重复结构域与磷酸肌醇相互作用蛋白1(WIPI1)的表达水平。结果与Ctrl组比较,LPS组小鼠肺组织病理学损伤加重,肺损伤评分、W/D值及细胞凋亡指数升高,ROS、IL⁃6及TNF⁃α含量明显升高,TFE3表达及核转位增多,HO⁃1、GRASP65、RAB20、STX3A的表达水平增加,WIPI1、GM130表达水平减少(均P<0.05),Hemin组小鼠上述各指标差异无统计学意义(均P>0.05)。与LPS组比较,LPS+Hemin组小鼠肺组织病理学损伤减轻,肺损伤评分、W/D值及细胞凋亡指数下降,ROS、IL⁃6及TNF⁃α含量减少,TFE3表达及核转位减少,HO⁃1、WObjective To investigate the effect of heme oxygenase⁃1(HO⁃1)on the expression and nuclear translocation of bHLH leucine zipper transcription factor E3(TFE3)expression,and golgi stress response in mice with endotoxin⁃induced acute lung injury(ALI).Methods According to the random number table method,24 mice were divided into four groups(n=6):a blank control(Ctrl)group,an en⁃dotoxin[lipopolysaccharide(LPS)]⁃induced acute lung injury(LPS)group,an endotoxin⁃induced acute lung injury+HO⁃1 agonist hemin(LPS+Hemin)group,and a Hemin group.Mice in the Ctrl group were injected with 0.5 ml of normal saline via the tail vein.Those in the LPS group were intravenously injected with LPS at 10 mg/kg via the tail vein.The LPS+Hemin group was intraperitoneally injected with hemin at 50 mg/kg,followed by intravenous injection of LPS at 10 mg/kg via the tail vein 1 h later to establish an ALI model.The Hemin group was in⁃traperitoneally injected with hemin at 50 mg/kg.Then,12 h after modeling,the mice were sacrificed and the lung tissues were harvested.Hematoxylin⁃eosin(H⁃E)staining was used to observe the histopathological changes of lung tissues and the pathological changes of lung tis⁃sues were scored.The lung wet to dry weight ratio(W/D ratio)was calculated.Cell apoptosis index was analyzed by terminal deoxynucleo⁃tidyl transferase⁃mediated dUTP⁃biotin nick end labeling(TUNEL).The levels of interleukin(IL)⁃6 and tumor necrosis factor(TNF)⁃αin lung tissues were determined by enzyme⁃linked immunosorbent assay(ELISA).The content of reactive oxygen species(ROS)was detected by flow cytometry.TFE3 nuclear translocation was observed by immunofluorescent staining.The levels of HO⁃1,TFE3,Golgi matrix pro⁃tein130(GM130),Golgi reassembly⁃stacking protein of 65 kDa(GRASP65),vesicular transportor⁃rab GTPase20(RAB20),syntaxin3A(STX3A),and WD repeat domain phosphoinositide⁃interacting protein1(WIPI1)were measured by Western blot.Results Compared with the Ctrl group,the LPS group showed aggravated lu

关 键 词：内毒素血症 急性肺损伤 血红素加氧酶⁃1 高尔基体 bHLH亮氨酸拉链转录因子E3 

分 类 号：R563.8[医药卫生—呼吸系统]