Heme oxygenase-1 attenuates endotoxin-induced acute lung injury through regulating bHLH leucine zipper transcription factor E3/Golgi stress response in mice
Objective 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 lungtis-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)-α in 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-tein 130(GM130),Golgi reassembly-stacking protein of 65 kDa(GRASP65),vesicular transportor-rab GTPase20(RAB20),syntaxin3A(STX3A),and WD repeat domain phosphoinositide-interacting protein 1(WIPI1)were measured by Western blot.Results Compared with the Ctrl group,the LPS group showed aggravated lung injury,and increases in lung injury scores,W/D values,cell apoptosis index,the contents of ROS,the concentrations of IL-6 and TNF-α,TFE3 expression and nuclear translocation,and the levels of HO-1,GRASP65,RAB20 and STX3A(all P＜0.05),as well as decreases in the levels of WIPI1 and GM 130(all P＜0.05).No statistical differenc-es were found as to the above-mentioned parameters in the Hemin group(all P＞0.05).Compared with the LPS group,the LPS+Hemin group presented relieved lung injury,decreases in lung injury scores and W/D values,cell apoptosis index,the contents of ROS,the con-centrations of IL-6 and TNF-α,TFE3 expression and nuclear translocation,and the levels of HO-1,WIPI1 and GM130(all P＜0.05),as well decreases in the levels of GRASP65,RAB20 and STX3A(all P＜0.05).Conclusions HO-1 attenuates endotoxin-induced ALI in mice,which be related to its regulation of TFE3 expression,nuclear translocation and Golgi stress response.
NETL
NSTL
万方数据
