黄芪多糖对稳定期慢性阻塞性肺疾病患者外周单核巨噬细胞Toll样受体4/核因子-κB通路的影响

摘要

目的 探讨黄芪多糖对稳定期慢性阻塞性肺疾病(COPD)患者外周单核巨噬细胞Toll样受体4(TLR4)/核因子(NF)-κB通路的影响.方法 选取稳定期COPD患者6例为试验组、肺功能正常的健康者6例为对照组,抽取受试者外周血,分离外周血单个核细胞、诱导形成单核细胞源巨噬细胞.将试验组分为COPD组、脂多糖组(10μg/ml)、黄芪多糖组(浓度为200、100、50μg/ml)、联合组(脂多糖10μg/ml+黄芪多糖200μg/ml、脂多糖10μg/ml+黄芪多糖100μg/ml、脂多糖10μg/ml+黄芪多糖50μg/ml),对照组和COPD组无药物干预.采用酶联免疫吸附试验检测细胞上清液中肿瘤坏死因子(TNF)-α、IL-6、基质金属蛋白酶(MMP)-9、前列腺素(PG)E2浓度,实时荧光定量聚合酶链反应检测单核细胞源巨噬细胞中TLR4、NF-κB mRNA的表达并进行比较.结果 对各组TNF-α、IL-6、MMP-9、PGE2和TLR4、NF-κB mRNA表达的比较,COPD组高于对照组;脂多糖组高于COPD组;黄芪多糖组低于COPD组;联合组低于脂多糖组(P均<0.01);黄芪多糖组内和联合组内随着黄芪多糖浓度的降低,上述指标依次升高(P<0.05).结论 稳定期COPD患者TLR4/NF-κB通路活化及其下游炎症因子分泌增加,黄芪多糖可抑制其TLR4/NF-κB通路、减少炎症因子分泌而发挥抗炎作用.%Objective To explore the effects of astragalus polysaccharide(APS) on Toll like receptor 4(TLR4)uclear factor kappa B(NF-κB) pathway of monocyte-derived macrophages from patients with stable chronic obstructive pulmonary disease(COPD).Methods Six patients with stable COPD were recruited as experimental group and six healthy people as control group.The mononuclear cells in peripheral blood were isolated and induced to macrophages.Monocyte-derived macrophages from control group were divided into COPD group,lipopolysaccharide(LPS) group(10μg/ml),APS groups(the concentrations were 200,100,50μg/ml) and combined group(including LPS 10μg/ml+APS 200μg/ml,LPS 10μg/ml+APS 100μg/ml and LPS 10μg/ml+APS 50μg/ml).Macrophages from control group and COPD group did not receive any drug intervention.Levels of Tumor necrosis factor(TNF)-α,IL-6,matrix metalloproteinase(MMP)-9 and prostaglandin(PG)E2 in culture supernatant were measured by ELISA and mRNA of TLR4 and NF-κB were measured by real-time PCR.Results The expression of TNF-α,IL-6,MMP-9,PGE2 in culture supernatant and mRNA of TLR4 and NF-κB were compared between groups,COPD group were higher than those in control group.LPS group were higher than those in COPD group.APS group were lower than those in COPD group.Combined group were lower than those in LPS group(P<0.01).Above-mentioned indexes increased with decrease of APS concentration in APS group and combined group(P<0.05).Conclusion TLR4/NF-κB pathway was activated and its downstream inflammatory cytokines increased in patients with stable COPD.APS can inhibit TLR4/NF-κB pathway and reduce the release of inflammatory cytokines and then plays an anti-inflammatory effect in stable COPD.