线粒体分裂抑制剂-1在香烟诱导小鼠气道炎症与氧化应激中的作用机制

Effect of mitochondria division inhibitor-1 on cigarette smoke-induced airway inflammation and oxidative stress in mouse

目的 探索线粒体分裂抑制剂-1(Mdivi-1)在香烟诱导小鼠气道炎症与氧化应激中的作用及相关的作用机制。方法 香烟暴露构建小鼠气道炎症与氧化应激动物模型,腹腔注射Mdivi-1进行干预,收集肺泡灌洗液进行细胞分类计数,ELISA检测肺泡灌洗液炎症因子TNF-α、IL-6及MMP-9水平,生化试剂盒测定肺组织匀浆氧化应激指标MDA含量及SOD的活性,HE染色评估肺组织病理改变。在PubChem数据库中检索Mdivi-1作用靶点基因,对基因进行GO和KEGG分析,Western blot检测小鼠肺组织Drp1和Mfn2的表达变化及相关富集的通路。结果 熏烟诱导小鼠气道中的炎症细胞浸润增加、气道上皮细胞过度增生、气道上皮增厚,增加肺泡灌洗液中总细胞、中性粒细胞、巨噬细胞数量及炎症因子TNF-α、IL-6及MMP-9水平,增强氧化应激反应,而Mdivi-1预处理可以减轻上述改变。生物信息分析显示Mdivi-1潜在作用基因100个,GO分析显示富集到线粒体结构调节、凋亡过程、线粒体自噬等生物过程,KEGG分析显示富集到的主要通路为Tnf、Mtor、Ampk、Adipocytokine及Nod-Like受体信号通路。Western blot显示Mdivi-1干预抑制香烟诱导小鼠肺组织NF-κB信号通路激活,下调香烟诱导的Drp1表达并上调Mfn2表达。结论 Mdivi-1缓解香烟诱导的小鼠气道炎症和氧化应激,其机制可能通过改善线粒体功能和抑制NF-κB信号通路活化实现,Mdivi-1有可能是治疗香烟诱导的气道炎症性疾病的新药物。

Objective This study aimed to investigate the effects of mitochondria division inhibitor-1(Mdivi-1)on cigarette smoke-induced airway inflammation and oxidative stress in mice and the possible mechanisms.Methods Mice were exposed to cigarette smoke to construct animal models of airway inflammation and oxidative stress,and midivi-1 was delivered by intraperitoneal injection. The bronchoalveolar lavage fluid(BALF)was collected for cell counting analysis. The levels of TNF-α,IL-6,MMP-9 in BALF were detected by ELISA. The levels of oxidative stress,including MDA and the activity of SOD,in lung tissue homogenate,were determined via biochemical kits. HE staining was used to evaluate the pathological changes of lung tissues. The possible genes Mdivi-1affected were searched in the PubChem database,followed by GO and KEGG analysis. Western blot was used to determine the expression changes of Drp1 and Mfn2 in the mouse lung tissue,and related enriched signal pathway.Results Cigarette smoke induced increased inflammatory cell infiltration,airway epithelial hyperplasia,airway epithelial thickening,and increased the number of total cells,neutrophils,and macrophages in BALF. The levels of TNF-α,IL-6 and MMP-9 and oxidative stress were also increased,while pretreatment with Mdivi-1 could alleviate the above changes. 100 potential target genes of Mdivi-1 were retrieved by bioinformatic analysis,and GO analysis showed that they were enriched in biological processes of regulation of mitochondrion organization,apoptosis,and mitophagy,and KEGG analysis identified enriched signal pathways of Tnf,Mtor,Ampk,Adipocytokine and Nod-Like receptor. Western blot showed that pretreatment with Mdivi-1 could significantly inhibited the activation of NF-κB signaling pathway,down-regulated Drp1expression and up-regulated Mfn2 expression in cigarette smoke-induced lung tissue of mice.Conclusion Mdivi-1 could alleviate airway inflammation and oxidative stress induced by cigarette smoke in mice,the mechanism of which might be achieved by improving mitochondrial function and inhibiting NF-κB signaling pathway. Thus,Mdivi-1 might be a novel drug for cigarette smoke-induced airway diseases.