膜联蛋白A1促进肺成纤维细胞炎症反应及胶原合成的研究

Effects of Annexin A1 on inflammatory response and collagen deposition of lung fibroblast

目的探讨膜联蛋白A1(Annexin A1)对肺成纤维细胞炎症反应、增殖及胶原沉积的作用。方法分离小鼠原代肺成纤维细胞,利用不同浓度的Annexin A1干预肺成纤维细胞24 h或48 h后,检测炎症因子、细胞增殖及胶原沉积表达。将C57/BL6小鼠分为对照组、Annexin A1干预组,每组6~8只。Annexin A1干预组每只小鼠气管滴注1μg/50μL Annexin A1重组因子,对照组每只小鼠气道滴注50μL生理盐水,连续干预7 d后取检。检测肺组织炎症因子、Ⅲ型胶原(CollagenⅢ)及气道平滑肌肌动蛋白(α-SMA)的表达。结果与对照组比较,Annexin A1能促进肺成纤维细胞分泌IL-6、IL-8等炎症因子(均P<0.05),促进成纤维细胞的增殖,CollagenⅢ合成和α-SMA表达增加(均P<0.05)。与对照组小鼠比较,Annexin A1干预组小鼠的肺泡灌洗液细胞总数[(2.91±0.26)×10^(4)/mL vs.(7.03±0.48)×10^(4)/mL,t=7.432,P=0.008]及中性粒细胞数目明显升高[(0.12±0.04)×10^(4)/mL vs.(1.01±0.05)×10^(4)/mL,t=13.810,P<0.001],炎症因子、胶原沉积及平滑肌表达也显著增加(均P<0.05)。结论 Annexin A1具有诱导肺成纤维细胞的炎症反应、增殖及胶原沉积作用,提示可能在气道重构发病中起重要作用。

Objective To investigate the effects of Annexin A1 on inflammatory response,proliferation and collagen deposition of pulmonary fibroblasts.Methods The primary lung fibroblasts of C57/BL6 mice were isolated.After 24 or 48 hours of intervention with different concentrations of Annexin A1,the expression of inflammatory factors,cell proliferation and collagen deposition were detected.C57/BL6 mice were randomly divided into control group and Annexin A1 intervention group according to the random number table method,with 6-8 mice in each group.Each mouse in the Annexin A1 intervention group was instilled with mouse recombinant Annexin A1(1μg/50μL)into the trachea,and each mouse in the control group was instilled with 50μL saline.After continuous intervention for 7 days,mice were sacrificed for measurement.Inflammatory factors,collagenⅢand airway smooth muscle actin(α-SMA)were detected.Results Inflammatory cytokines(IL-6,IL-8),proliferation,collagenⅢ,andα-SMA were significantly increased in the Annexin A1 intervention group compared with the control group(all P<0.05).Compared with control mice,the total number of cells in the alveolar lavage fluid of the Annexin A1 intervention group[(2.91±0.26)×10^(4)/mL vs.(7.03±0.48)×10^(4)/mL,t=7.432,P=0.008]and the number of neutrophils[(0.12±0.04)×10^(4)/mL vs.(1.01±0.05)×10^(4)/mL,t=13.810,P<0.001]were significantly increased,and the expression of inflammatory cytokines,collagen deposition,and smooth muscle were also significantly increased(all P<0.05).Conclusion Annexin A1 induces inflammation,proliferation and collagen deposition in the lung fibroblasts,suggesting that Annexin A1 maybe involved in the pathogenesis of airway remodeling.