Regulation of LPS-mediated inflammation in vivo and in vitro by the thiol antioxidant Nacystelyn
Regulation of LPS-mediated inflammation in vivo and in vitro by the thiol antioxidant Nacystelyn
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DOI：
10.1152/ajplung.00329.2003
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发表时间：
2004-06-01
影响因子：
4.9
通讯作者：
MacNee, W
中科院分区：
文献类型：
--
作者：
Antonicelli, F;Brown, D;MacNee, W
在各种肺部疾病中，支气管肺泡灌洗液中存在促炎细胞因子水平升高。氧化还原敏感性转录因子如NF-κ B调节这些细胞因子的基因转录。因此，我们研究了一种新的巯基抗氧化剂化合物，Nacystelyn（NAL），在人巨噬细胞衍生的细胞系（THP-1）的IL-8调节的影响。与对照水平相比，LPS（10 μ g/ml）增加IL-8的释放。通过与NAL（1和5mM）共孵育来抑制该LPS活化。预处理放线菌酮或冈田酸，蛋白质合成，丝氨酸/苏氨酸磷酸酶抑制剂，分别没有修改由NAL引起的IL-8释放的抑制。与对照组相比，LPS处理后NF-κ B和C/EBP DNA结合增加，与NAL共处理抑制这种效应。激活蛋白（AP）-1 DNA结合不受影响。当细胞与NAL共处理时，LPS处理细胞的条件培养基产生的增强的中性粒细胞趋化性被抑制。研究了NAL抑制对IL-8表达的选择性。LPS处理的THP-1细胞也具有较高水平的TNF-α、转化生长因子（TGF）-β 1和-3、MIP-1 α和-β以及RANTES基因表达。然而，只有LPS诱导的IL-8和TGF-β 1的表达被NAL抑制。NAL的抗炎作用在体内得到证实，如通过将NAL滴注到肺中后LPS诱导的中性粒细胞向肺的募集减少所示。我们的研究表明，NAL在体外和体内具有抗炎特性，因此可能在肺部炎症中具有治疗作用，并且与其他抗氧化剂相比具有优势，因为它可以通过吸入给药。
Increased levels of proinflammatory cytokines are present in bronchoalveolar lavage fluid in various lung diseases. Redox-sensitive transcription factors such as NF-kappaB regulate gene transcription for these cytokines. We therefore studied the effect of a new thiol antioxidant compound, Nacystelyn (NAL), on IL-8 regulation in a human macrophage-derived cell line (THP-1). LPS (10 mug/ml) increased IL-8 release compared with control levels. This LPS activation was inhibited by coincubation with NAL (1 and 5 mM). Pretreatment with cycloheximide or okadaic acid, protein synthesis, and serine/threonine phosphatase inhibitors, respectively, did not modify inhibition of IL-8 release caused by NAL. NF-kappaB and C/EBP DNA binding were increased after LPS treatment compared with control, an effect inhibited by cotreatment with NAL. Activator protein (AP)-1 DNA binding was unaffected. The enhanced neutrophil chemotaxis produced by conditioned media from LPS-treated cells was inhibited when cells were cotreated with NAL. The selectivity of NAL inhibition upon IL-8 expression was studied. LPS-treated THP-1 cells also had higher levels of TNF-alpha, transforming growth factor (TGF)-beta1 and -3, MIP-1alpha and -beta, and RANTES gene expression. However, only LPS-induced IL-8 and TGF-beta1 expressions were inhibited by NAL. An anti-inflammatory effect of NAL was confirmed in vivo as shown by a reduction in LPS-induced neutrophil recruitment to the lungs following instillation of NAL into the lungs. Our studies demonstrate that NAL has anti-inflammatory properties in vitro and in vivo, may therefore have a therapeutic role in lung inflammation, and has the advantage over other antioxidant agents in that it may be administrated by inhalation.