Evidence of a defective thiol status of alveolar macrophages from COPD patients and smokers
Evidence of a defective thiol status of alveolar macrophages from COPD patients and smokers
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DOI：
10.1016/s0891-5849(00)00424-x
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发表时间：
2000-12-01
影响因子：
7.4
通讯作者：
Welte, T
中科院分区：
文献类型：
--
作者：
Täger, M;Piecyk, A;Welte, T
在越来越多的肺部疾病与细胞内的巯基，主要是谷胱甘肽的损失，是假定。因此，不同活细胞内硫醇的定量测量是所有肺免疫细胞（包括肺泡巨噬细胞（AM））的细胞功能和防御能力的可能代谢参数，这些细胞受到氧化应激的高度损害。在这项研究中，使用荧光染料缀合的氯甲基衍生物（5-氯甲基荧光素二乙酸酯，CMFDA）在流式细胞术中测定细胞硫醇含量。使用谷胱甘肽定量的生化技术在体外评价该程序。基于这种方法，与非吸烟者/非COPD组相比，从吸烟者和慢性阻塞性肺病（COPD）患者的支气管肺泡灌洗（BAL）中获得的AM显示出显着的硫醇缺乏。吸烟者和COPD患者的AM细胞巯基表达仅为对照组的50%和53%。在吸烟者（+）/COPD+组中检测到最低硫醇浓度（对照组的47%）。这种细胞内巯基缺乏与肺功能（FEV 1，PaO 2）降低显著相关。关于免疫细胞的严格调节的硫醇代谢，这些结果意味着硫醇缺陷AM中功能紊乱的发作。通过流式细胞术测定从BAL获得的AM的细胞硫醇含量，提供了一种简单可靠的工具来监测专注于细胞硫醇状态稳定的治疗措施的效果。(C)2000 Elsevier Science Inc.
In increasing numbers of pulmonary diseases an association with a loss of intracellular thiols, mainly glutathione, is postulated. Therefore, the quantitative measurement of thiols within different viable cells is a possible metabolic parameter for cellular function and defense capacity of all pulmonary immune cells including alveolar macrophages (AM), that are highly compromised by oxidative stress. In this study the cellular thiol content was determined using fluorochrom conjugated chloromethyl derivatives (5-chloromethylfluorescein diacetate, CMFDA) in flow cytometry. The procedure was evaluated in vitro using biochemical techniques for glutathione quantification. Based on this approach, AM obtained from bronchoalveolar lavage (BAL) of smokers and patients with chronic obstructive pulmonary disease (COPD) showed a significant thiol deficiency compared to a nonsmoker/non-COPD group. The cellular thiol expression of AM from smokers and COPD patients reached only 50 and 53% of the control group. Lowest thiol concentrations (47% of control) were detected within the smoker(+)/COPD+ group. This intracellular thiol deficiency significantly correlated with reduced lung function (FEV1, PaO2). With regard to the tightly regulated thiol metabolism of immune cells, these results imply the onset of functional disturbances in thiol deficient AM. The determination of the cellular thiol content of AM, obtained from BAL by flow cytometry, presents a simple and reliable tool to monitor the effect of therapeutic measures focusing on the stabilization of the cellular thiol status. (C) 2000 Elsevier Science Inc.