【摘要】 目的:探讨PM2.5刺激对慢性阻塞性肺疾病急性加重期(acute exacerbation chronic obstructive pulmonary disease,AECOPD)及哮喘-慢性阻塞性肺疾病重叠(asthma-chronic obstructive pulmonary disease overlap,ACO)患者血清孵育的肺泡巨噬细胞(MH-S)的影响及PM2.5致巨噬细胞炎症与SIRT1/NF-κB信号通路的关系。方法:首先将细胞分为对照组、PM2.5组、AECOPD组、ACO组、PM2.5+AECOPD组和PM2.5+ACO组,培养MH-S细胞6 h,收集细胞上清液,ELISA检测不同组之间白细胞介素6(IL-6)和IL-10浓度的差异。然后,为探究PM2.5致巨噬细胞炎症与SIRT1/NF-κB通路的关系,进一步将MH-S细胞分为对照组、PM2.5组、PM2.5+SRT2104组和PM2.5+EX527组,检测细胞沉默信息调节因子2相关酶1(silent information regulator 2-related enzymes 1,SIRT1)、NF-κB P65、p-P65、IκB和p-IκB蛋白的水平,以及细胞上清液中IL-6和IL-10的浓度。结果:与对照组相比,PM2.5组、AECOPD组、ACO组、PM2.5+AECOPD组和PM2.5+ACO组的IL-6浓度均升高,IL-10浓度均降低,其中ACO组较AECOPD组的变化更明显(P<0.05)。与PM2.5+AECOPD组相比,PM2.5+ACO组的IL-6升高更明显(P<0.05)。PM2.5刺激细胞12 h后,SIRT1、IκB和p-IκB的蛋白水平减少,P65和p-P65的蛋白水平增加(P<0.05);加入SIRT1激活剂SRT2104,SIRT1的表达升高,p-P65蛋白水平降低,IL-6浓度降低,IL-10浓度升高(P<0.05);加入SIRT1抑制剂EX527,SIRT1的表达减少,P65和p-P65蛋白水平升高,IL-6浓度升高,IL-10浓度降低(P<0.05)。结论:PM2.5可以通过SIRT1/NF-κB信号通路致MH-S细胞炎症,且PM2.5对ACO患者的致炎作用可能高于AECOPD患者。更多还原

【Abstract】 AIM:To investigate the effects of PM2.5,and sera from acute exacerbation of chronic obstructive pulmonary disease(AECOPD)and asthmatic-COPD overlap(ACO)patients on MH-S cell inflammation,and the relationship between macrophage inflammation stimulated by PM2.5 and SIRT1/NF-κB signaling pathway.METHODS:The MH-S cells were divided into control group,PM2.5 group,AECOPD group,ACO group,PM2.5+AECOPD group,PM2.5+ACO group.The cells were cultured for 6 h,and the supernatant was collected.The concentration of interleukin-6(IL-6)and IL-10 in cell supernatant were measured by ELISA.The concentrations of IL-6 and IL-10 were compared between different groups.To further explore the relationship between PM2.5 induced macrophage inflammation and SIRT1/NF-κB pathway,the MH-S cells were divided into control group,PM2.5 group,PM2.5+SRT2104 group and PM2.5+EX527 group.The cells were stimulated for 12 h.The cell and cell supernatant were collected to detect the protein levels of SIRT1,NF-κB P65,p-P65,inhibitor of NF-κB(IκB)and p-IκB,and the concentrations of IL-6 and IL-10.RESULTS:Compared with control group,the concentration of IL-6 was increased and the IL-10 was decreased in PM2.5 group,AECOPD group,ACO group,PM2.5+AECOPD group and PM2.5+ACO group.Compared with AECOPD group,IL-6 was increased and IL-10 was decreased more obviously in ACO group(P<0.05).Compared with PM2.5+AECOPD group,the increase of IL-6 was more obvious(P<0.05),and no significant difference in IL-10 was observed in PM2.5+ACO group.After PM2.5 stimulation for 12 h,the expression of SIRT1 was decreased,the protein levels of IκB and pIκB were decreased,and the protein levels of P65 and p-P65 were increased(P<0.05).After adding SIRT1 activator SRT2104,the expression of SIRT1 was increased(P<0.05),p-P65 level was decreased,IL-6 content was decreased,and IL-10 level was increased(P<0.05).After adding SIRT1 inhibitor EX527,the expression of SIRT1 were decreased,the protein levels of P65 and p-p65 were increased,the concentration of IL-6 was increased and the level of IL-10 was decreased(P<0.05).CONCLUSION:PM2.5 induces MH-S cell inflammation through SIRT1/NF-κB signaling pathway.PM2.5 may have a stronger proinflammatory effect on ACO patients than on AECOPD patients.更多还原