外周血单个核细胞干扰素调节因子-1和去整合素金属蛋白酶-8表达对支气管哮喘的调控作用

Regulatory roles of interferon regulatory factor-1 and a disintegrin and metalloproteinase-8 in bronchial asthma

目的观察支气管哮喘患者外周血单个核细胞干扰素调节因子-1(interferon regulatory factor-1, IRF-1)和去整合素金属蛋白酶-8(a disintegrin and metalloproteinase-8, ADAM-8)表达变化,探讨其与肺功能、血清总免疫球蛋白(immunoglobin, Ig)E、外周血嗜酸性粒细胞(eosnophils, EOS)百分比(EOS%)的相关性。方法 60例支气管哮喘患者为观察组,其中轻度持续组23例,中度持续组19例,重度持续组18例;同期体检健康者60例为对照组。观察组给予噻托溴铵粉雾剂+沙美特罗替卡松气雾剂治疗,采用哮喘控制测试(asthma control test, ACT)问卷评估哮喘控制情况。应用肺功能仪测定观察组入院时第1秒用力呼气容积(the forced expiratory volume in one second, FEV_(1))占预计值百分比(FEV_(1)%pred),计算FEV_(1)/用力肺活量(forced vital capacity, FVC)比值;入院次日采集血液测定外周血EOS%、血清总IgE水平;采用实时荧光定量PCR法检测外周血单个核细胞IRF-1 mRNA、ADAM-8 mRNA相对表达量;采用Western blot法检测外周血单个核细胞ADAM-8、IRF1蛋白相对表达量;并与对照组进行比较。Pearson相关法分析支气管哮喘患者外周血单个核细胞IRF-1、ADAM-8表达与FEV_(1)%pred、FEV_(1)/FVC、血清总IgE、外周血EOS%的相关性;多重线性回归分析影响支气管哮喘患者ACT评分的因素。结果观察组FEV_(1)%pred[(82.23±5.25)%]、FEV_(1)/FVC(86.41±7.92)均低于对照组[(97.24±10.75)%、95.52±9.38](P<0.05),血清总IgE[(670.31±86.59) ku/L]、外周血EOS%[(5.89±0.92)%]均高于对照组[(376.89±78.75)ku/L、(2.90±0.77)%](P<0.05),外周血单个核细胞IRF-1 mRNA(1.76±0.39)、ADAM-8 mRNA(3.66±0.88)、IRF-1蛋白(1.98±0.66)、ADAM-8蛋白(2.27±0.75)相对表达量均高于对照组(1.02±0.13、0.97±0.14、0.92±0.25、1.08±0.22)(P<0.05)。轻、中、重度持续组FEV_(1)%pred[(89.72±7.95)%、(68.59±7.92)%、(49.75±8.36)%]、FEV_(1)/FVC(90.72±10.58、75.46±10.62、63.72±8.47)、ACT评分[(22.15±2.77)、(17.51±2.22)、(10.25±1.98)分]依次降低(P<0.05),血清总IgE水平[(422.58±71.29)、(603.26±89.31)、(713.45±100.27)ku/L]、外周血EOS%[(4.88±0.79)%、(5.52±0.81)%、(6.31±0.85)%]及外周血单个核细胞IRF-1 mRNA(1.34±0.13、1.85±0.16、2.18±0.21)、ADAM-8 mRNA(2.77±0.39、3.76±0.59、4.69±0.33)、IRF-1蛋白(1.37±0.26、2.12±0.47、2.61±0.48)、ADAM-8蛋白(1.63±0.35、2.28±0.48、3.08±0.52)相对表达量依次升高(P<0.05)。Pearson相关分析结果显示,支气管哮喘患者外周血单个核细胞ADAM-8、IRF-1 mRNA及蛋白表达与外周血EOS%、血清总IgE水平呈正相关(P<0.05),与FEV_(1)%pred、FEV_(1)/FVC呈负相关(P<0.05)。多重线性回归分析结果显示,外周血单个核细胞IRF-1 mRNA(β=-0.412,95%CI:-9.314~-2.066,P=0.003)、ADAM-8 mRNA(β=-0.263,95%CI:-2.260~-0.124,P=0.029)是支气管哮喘患者ACT评分的影响因素。结论支气管哮喘患者外周血单个核细胞ADAM-8、IRF-1表达上调,且随哮喘症状加重、肺功能下降而增高,有可能成为支气管哮喘新的治疗靶点。

Objective To observe the changes of interferon regulatory factor-1(IRF-1)and a disintegrin and metalloproteinase-8(ADAM-8)in peripheral blood mononuclear cells in patients with bronchial asthma(BA)and to investigate their correlations with lung function,total serum immunoglobin E(IgE)and peripheral blood eosinophils(EOS)percentage(EOS%).Methods In 60 BA patients(observation group),there were 23 patients with mild persistent BA(mild persistent BA group),19 patients with moderate persistent BA(moderate persistent BA group),and18 patients with severe persistent BA(severe persistent BA group).Another 60 healthy volunteers were as controls(control group).Observation group received Tiotropium Bromide Inhalation Powder+Salmeterol Xinafoate and Fluticasone Propionate Aerosol,and was assessed the control of BA by using Asthma Control Test(ACT)questionnaire.In observation group,the forced expiratory volume in one second(FEV_(1))in percentage of the predicted value(FEV_(1)%pred)and FEV_(1)/forced vital capacity(FVC)were detected at admission,the EOS%and total IgE level were detected by day 1 after admission;the relative expressions of IRF-1 mRNA and ADAM-8 mRNA were detected by real-time fluorescence quantitative PCR,the relative expressions of IRF-1 and ADAM-8 proteins were detected by Western blot,and all above indexes were compared with control group.Pearson correlation method was used to analyze the correlations of the expressions of IRF-1 and ADAM-8 in peripheral blood mononuclear cells with FEV_(1)%pred,FEV_(1)/FVC,total IgE level and EOS%.Multivariate linear regression analysis was used to assess the influencing factors of ACT.Results FEV_(1)%pred and FEV_(1)/FVC were lower in observation group((82.23±5.25)%,86.41±7.92)than those in control group((97.24±10.75)%,95.52±9.38)(P<0.05),EOS%and total IgE level were higher in observation group((5.89±0.92)%,(670.31±86.59)ku/L)than those in control group((2.90±0.77)%,(376.89±78.75)ku/L)(P<0.05),and the relative expressions of IRF-1 mRNA,ADAM-8 mRNA,IRF-1 protein and ADAM-8 protein were higher in observation group(1.76±0.39,3.66±0.88,1.98±0.66,2.27±0.75)than those in control group(1.02±0.13,0.97±0.14,0.92±0.25,1.08±0.22)(P<0.05).FEV_(1)%pred((89.72±7.95)%,(68.59±7.92)%,(49.75±8.36)%),FEV_(1)/FVC%(90.72±10.58,75.46±10.62,63.72±8.47)and ACT score(22.15±2.77,17.51±2.22,10.25±1.98)deceased gradually in turn in mild,moderate and severe persistent BA groups(P<0.05),while total IgE((422.58±71.29),(603.26±89.31),(713.45±100.27)ku/L),EOS%((4.88±0.79)%,(5.52±0.81)%,(6.31±0.85)%),IRF-1 mRNA(1.34±0.13,1.85±0.16,2.18±0.21),ADAM-8 mRNA(2.77±0.39,3.76±0.59,4.69±0.33),IRF-1 protein(1.37±0.26,2.12±0.47,2.61±0.48)and ADAM-8 protein(1.63±0.35,2.28±0.48,3.08±0.52)increased gradually in turn in mild,moderate and severe persistent BA groups(P<0.05).Pearson correlation analysis showed that ADAM-8 and IRF-1 mRNAs and proteins were positively correlated with EOS%and total IgE level(P<0.05),and negatively correlated with FEV_(1)%pred and FEV_(1)/FVC(P<0.05).Multivariate linear regression analysis showed that IRF-1 mRNA(β=-0.412,95%CI:-9.314 to-2.066,P=0.003)and ADAM-8 mRNA(β=-0.263,95%CI:-2.260 to-0.124,P=0.029)were the influnencing factors of ACT score.Conclusion The expressions of ADAM-8 and IRF-1 in peripheral blood mononuclear cells are up-regulated,and increase with the aggravation of BA and lung function.Both of ADAM-8 and IRF-1 would be the new therapeutic targets for BA.