西乐葆对重症肺炎小鼠ANXA1/FPR2通路及炎症反应的影响

Effects of Celecoxib on ANXA1/FPR2 pathway and inflammatory response in mice with severe pneumonia

目的:探究西乐葆对重症肺炎小鼠炎症反应及肺组织中膜联蛋白A1(ANXA1)/N-甲酰肽受体(FPR2)通路的影响。方法:采用肺炎克雷伯杆菌感染构建重症肺炎小鼠模型。选用BALB/c小鼠120只,随机分为对照组、模型组、西乐葆低剂量组(1.5 mg/kg)、西乐葆中剂量组(3 mg/kg)、西乐葆高剂量组(6 mg/kg)和地塞米松组,每组20只。分离并计数肺泡灌洗液(BALF)中的炎症细胞;测定小鼠肺指数和肺组织湿重/干重(W/D);ELISA法检测肺组织中髓过氧化物酶(MPO)活性和IL-6、IL-1β、TNF-α水平;HE染色检测肺组织病理学变化;TUNEL法检测肺组织细胞凋亡情况;Western blot检测肺组织ANXA1、FPR2蛋白水平。结果:与对照组相比,模型组小鼠肺组织结构破坏严重,肺泡大小不等,肺泡间隔增厚,可见明显充血,并伴有大量炎症细胞浸润,小鼠肺指数、细胞凋亡率、W/D、MPO活性、嗜酸性粒细胞、中性粒细胞、淋巴细胞、巨噬细胞数量及IL-6、IL-1β、TNF-α水平显著升高(P<0.05),ANXA1、FPR2蛋白表达水平显著降低(P<0.05);与模型组相比,西乐葆各剂量组及地塞米松组小鼠肺泡壁和肺泡间隔结构得到一定程度的修复,炎症细胞浸润减少,小鼠肺指数、细胞凋亡率、W/D、MPO活性、嗜酸性粒细胞、中性粒细胞、淋巴细胞、巨噬细胞数量及IL-6、IL-1β、TNF-α水平显著降低(P<0.05),ANXA1、FPR2蛋白表达水平显著升高(P<0.05)。结论:西乐葆可明显促进ANXA1/FPR2通路表达,减轻重症肺炎小鼠的炎症反应。

Objective:To investigate effects of Celecoxib on inflammatory response and Annexin A1(ANXA1)/N-formyl peptide receptor 2(FPR2)pathway in lung tissue of mice with severe pneumonia.Methods:Severe pneumonia mice model was established by Klebsiella pneumoniae infection.Total 120 BALB/c mice were randomly divided into control group,model group,Celecoxib low-dose group(1.5 mg/kg),Celecoxib medium-dose group(3 mg/kg),Celecoxib high-dose group(6 mg/kg)and Dexamethasone group,with 20 mice in each group.Inflammatory cells in bronchoalveolar lavage fluid(BALF)were isolated and counted;lung index and ratio of wet weight to dry weight(W/D)were measured;activity of myeloperoxidase(MPO)and levels of IL-6,IL-1βand TNF-αwere detected by ELISA;HE staining was used to detect pathological changes of lung tissue;TUNEL method was used to detect apoptosis of lung tissue;protein levels of ANXA1 and FPR2 were detected by Western blot.Results:Compared with control group,lung tissue structure of model group was seriously damaged,alveolar size was different,and alveolar septum was thickened,obvious hyperemia was observed,with a large number of inflammatory cells infiltrated,lung index,apoptosis rate,W/D,MPO activity,numbers of eosinophils,neutrophils,lymphocytes and macrophages,and levels of IL-6,IL-1βand TNF-αwere significantly increased(P<0.05),and expression levels of ANXA1 and FPR2 protein were significantly decreased(P<0.05).Compared with model group,alveolar wall and alveolar septum structure of Celecoxib groups and Dexamethasone group mice were repaired to a certain extent,and the infiltration of inflammatory cells was reduced,lung index,apoptosis rate,W/D,MPO activity,numbers of eosinophils,neutrophils,lymphocytes and macrophages,and levels of IL-6,IL-1βand TNF-αwere significantly decreased(P<0.05),and expression levels of ANXA1 and FPR2 protein were significantly increased(P<0.05).Conclusion:Celecoxib can significantly promote expression of ANXA1/FPR2 pathway and reduce inflammatory response in mice with severe pneumonia.