ORMDL3通过内质网应激介导的细胞自噬在中性粒细胞哮喘中的作用机制研究  

Role and mechanism of ORMDL3 in neutrophil asthma through endoplasmic reticulum stress-mediated autophagy

作　　者：王龙梅[1] 孙永显[1] 张新鹃[1] 陈晓艳[1] WANG Longmei;SUN Yongxian;ZHANG Xinjuan;CHEN Xiaoyan(Western Medicine Teaching Department,Shandong College of Traditional Chinese Medicine,Yantai 264199,China)

机构地区：[1]山东中医药高等专科学校,西医教学部,山东烟台264199

出　　处：《中国比较医学杂志》2022年第3期46-53,共8页Chinese Journal of Comparative Medicine

基　　金：山东省卫计委课题项目(2018020807)。

摘　　要：目的 研究ORMDL3通过内质网应激(ERS)介导的细胞自噬在中性粒细胞哮喘中的作用机制。方法 成年雄性SD大鼠随机分为对照组、模型组、模型+NC-siRNA组、模型+ORMDL3-siRNA组、溶剂对照组、模型+溶剂组、模型+NC-siRNA+溶剂组、模型+ORMDL3-siRNA+溶剂组、模型+ORMDL3-siRNA+激动剂组,采用脂多糖+卵白蛋白联合致敏的方式建立中性粒细胞哮喘模型,经气道注射ORMDL3-siRNA慢病毒或阴性对照(NC)-siRNA慢病毒,腹腔注射ERS激动剂毒胡萝卜素或溶剂。检测气道功能0.2 s用力呼气容积(FEV0.2)/用力肺活量(FVC)以及呼气峰流速(PEF),肺组织病理改变,ORMDL3、ERS标志基因PERK、IRE1α、ATF6及自噬标志基因Beclin-1、LC3的表达水平。结果 模型组大鼠的FEV0.2/FVC、PEF水平低于对照组,肺组织中ORMDL3、PERK、IRE1α、ATF6、Beclin-1、LC3-II/LC3-I的表达水平高于对照组(P<0.05);模型+ORMDL3-siRNA组大鼠的FEV0.2/FVC、PEF水平高于模型+NC-siRNA组,肺组织中ORMDL3、ATF6、Beclin-1、LC3-II/LC3-I的表达水平低于模型+NC-siRNA组(P<0.05)。PERK、IRE1α的表达水平与模型+NC-siRNA组比较无差异(P>0.05);模型+ORMDL3-siRNA+激动剂组大鼠的FEV0.2/FVC、PEF水平低于模型+ORMDL3-siRNA+溶剂组,肺组织中Beclin-1、LC3-II/LC3-I的表达水平高于模型+ORMDL3-siRNA+溶剂组(P<0.05)。结论 在中性粒细胞哮喘的发病中,ORMDL3通过ERS的ATF6通路激活细胞自噬是可能的分子机制,阻断ORMDL3是治疗中性粒细胞哮喘的可能靶点。Objective To investigate the role and mechanism of ORMDL3 in neutrophil asthma through endoplasmic reticulum stress(ERS)-mediated autophagy. Methods Adult male SD rats were randomly divided into control, model, model+NC siRNA, model+ORMDL3 siRNA, solvent control, model+solvent control, model+NC siRNA group+solvent control, model+ORMDL3 siRNA+solvent control and model+ORMDL3 siRNA+agonist groups. The neutrophil asthma model was established by lipopolysaccharide and ovalbumin sensitization. An ORMDL3 siRNA lentivirus or negative control(NC)-siRNA lentivirus was administered through the airway and ERS agonist, carotene, or solvent was injected intraperitoneally. Airway function, 0.2 s forced expiratory volume(FEV0.2)/forced vital capacity(FVC), peak expiratory flow(PEF), pathological changes of lung tissue and the expression levels of ORMDL3, ERS marker genes PERK, IRE1α, ATF6 and autophagy marker genes Beclin-1 and LC3 were analyzed. Results FEV0.2/FVC and PEF levels of the model group were lower than those of the control group and the expression levels of ORMDL3, PERK, IRE1α, ATF6, Beclin-1 and LC3-II/LC3-I in lung tissue were higher than those of the control group(P<0.05). FEV0.2/FVC and PEF levels of the model+ORMDL3-siRNA group were higher than those of the model+NC-siRNA group and the expression levels of ORMDL3, ATF6, Beclin-1 and LC3-II/LC3-I in lung tissue were lower than those of the model+NC-siRNA group(P<0.05). The expression levels of PERK and IRE1α showed no difference compared with the model+NC-siRNA group(P>0.05). FEV0.2/FVC and PEF levels in the model+ORMDL3 siRNA+agonist group were lower than those in the model+ORMDL3 siRNA+solvent group and the expression levels of Beclin-1 and LC3-II/LC3-I in lung tissue were higher than those in the control group(P<0.05). Conclusions In the pathogenesis of neutrophil asthma, ORMDL3-induced autophagy through ERS ATF6 pathway is a possible molecular mechanism. Blocking ORMDL3 is a possible treatment method for neutrophil asthma.

关 键 词：中性粒细胞哮喘 ORMDL3 内质网应激 自噬 分子机制 

分 类 号：R-33[医药卫生]