【作者】 杨蕾；

【导师】 吴大玮；

【作者基本信息】 山东大学 ， 内科学， 2007， 硕士

【摘要】 目的观察地塞米松(Dex)、茶碱(Theo)、N-乙酰半胱氨酸(NAC)单药或联合干预对慢性阻塞性肺疾病(COPD)患者肺泡巨噬细胞(AM)细胞因子IL-8与TNF-α的分泌、核因子-κB(NF-κB)的表达及组蛋白去乙酰化酶(HDAC)活性的影响，探讨单药与联合用药对LPS刺激后COPD肺泡巨噬细胞的抗炎作用是否存在差异，为COPD的抗炎治疗提供理论依据。方法分离培养吸烟COPD患者，吸烟肺功能正常者及正常对照者的肺泡巨噬细胞，将每位受试者的AM分为LPS组(LPS刺激)、Dex组(LPS刺激+Dex干预)、Theo组(LPS刺激+Theo干预)、NAC组(LPS刺激+NAC干预)，及联合干预组(LPS刺激+Dex+Theo+NAC)共5组。分别采用酶联免疫吸附法检测细胞培养上清中IL-8、TNF-α的含量，免疫细胞化学法检测肺泡巨噬细胞NF-κB的表达及色度法测定核蛋白中HDAC的活性。结果①经LPS刺激后，COPD组及吸烟肺功能正常组AM分泌IL-8与TNF-α较正常对照组增加(P＜0.01或0.05)；Dex和NAC干预均可使COPD组及吸烟肺功能正常组AM的IL-8和TNF-α分泌下降(P＜0.01)；联合干预组抑制IL-8与TNF-α分泌的作用较单一药物更强(P＜0.001)。②COPD组及吸烟肺功能正常组NF-κBp65蛋白阳性细胞表达与正常对照组比较明显升高(P＜0.05)；与LPS刺激组相比，药物干预后的各组NF-κB蛋白阳性细胞表达降低(P＜0.05)；联合干预组NF-κB蛋白阳性细胞表达进一步降低(P＜0.01)。③COPD组、吸烟肺功能正常组较正常对照组，AM经LPS刺激后HDAC活性降低(P＜0.05)；COPD组与吸烟肺功能正常组AM经茶碱或NAC干预后HDAC活性显著增加(P＜0.01)；联合干预组HDAC的活性显著增加(P＜0.01)；Dex对HDAC活性无显著性影响。结论①低浓度茶碱、地塞米松、N-乙酰半胱氨酸单一或联合应用，可抑制COPD肺泡巨噬细胞IL-8和TNF-α分泌、NF-κB表达；②低浓度茶碱、N-乙酰半胱氨酸增加HDAC的活性；③联合干预的作用优于单一药物。更多还原

【Abstract】 Objective: To investigate the effects of individual and combinated administration of dexamethasone, theophylline, and N-acetyleysteine (NAC) on IL-8 and TNF-α release 、 expression of nuclear factor kappaB (NF- κB) and activity of histone deacetylace(HDAC) in Alveolar macrophages(AMs) from COPD patients.Methods: AM isolated and purified from normal subjects, healthy smokers, and COPD patients were randomly divided into five groups The LPS stimulation group(LPS stimulation), the theophylline group (LPS stimulation+Theo), the dexamethasone group ((LPS stimulation+Dex), the NAC group((LPS stimulation+ NAC) and the combination group with dexamethasone、 theophylline and NAC ((LPS stimulation+Dex +Theo+ NAQ.The IL-8 and TNF-α concentrations were determined by sandwich ELISA. The expression of NF-κB in AM was observed by immunohistochemistry technique. HDAC activity was measured in protein extracts from AM and expressed according to protein content.Results: Compared with normal ,the secretion of IL-8 and TNF-α was increased significantly in AM from smokers and COPD patients after LPS stimulation (P<0.01or0.05); Dexamethasone and NAC can reduce IL-8 and TNF-α release in AM from COPD patients and smokers (P<0.01); Combination group has more effect on inhibiting the secretion of IL-8 and TNF-α (P<0.001) .Compared with normal group, the expression of NF-κB was increased significantly in AM from smokers and COPD patients after LPS stimulation (P< 0.05 ). the expression of NF-κB was less in groups of drug settled than in LPS stimulating group(P<0.05),and decreased further in combination group(P<0.01).HDAC activity was significantly reduced in AM from COPD patients and smokers compared with normal subjects after LPS stimulation(P<0.05).Theophylline and NAC can increase HDAC activity in AM from COPD patients and smokers (P<0.01), while combination groups can increase more (P<0.01); .DEX has no effect on the activity of HDAC.Conclusion: The individual and combinated administration of low-doses theophylline, dexamethasone, and N-acetyleysteine can reduce the release of IL-8 and TNF-α, inhibit the expression of NF-κB; Low-doses theophylline and N-acetyleysteine can improve the activity of HDAC2; Combinated administration has better effect than individual in the treatmeant of COPD.更多还原