Carbonylation Caused by Cigarette Smoke Extract Is Associated with Defective Macrophage Immunity
Carbonylation Caused by Cigarette Smoke Extract Is Associated with Defective Macrophage Immunity
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DOI：
10.1165/rcmb.2010-0272oc
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发表时间：
2011-08-01
影响因子：
6.4
通讯作者：
Anderson, Gary P.
中科院分区：
文献类型：
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作者：
Bozinovski, Steven;Vlahos, Ross;Anderson, Gary P.
香烟烟雾中的氧化剂抑制病原体识别受体功能和吞噬功能，但这种抑制的分子基础尚不清楚。我们试图确定损害肺泡巨噬细胞功能的抑制机制。BALB/c小鼠急性暴露于4支香烟4h后，滴鼻内毒素(1 MU G)。小鼠的气道中性粒细胞和肿瘤坏死因子-α的表达显著降低。香烟烟雾提取物(CSE)对BALB/c来源的MH-S肺泡巨噬细胞具有类似的抑制作用。CSE还可抑制重组(R)肿瘤坏死因子-α(100 ng/ml)对炎症基因的诱导。由于这两条通路都汇聚在核因子-kappaB上，因此对ikappa Bα的降解和p65的磷酸化进行了评估，并显示CSE钝化了这两条通路。CSE还以还原型谷胱甘肽(GSH)可逆的方式抑制p38丝裂原活化蛋白激酶(MAPK)，从而阻断激活蛋白-1(AP-1)的活性。特异性Toll样受体(TLR)负性调节因子(细胞因子信号转导抑制因子-1[SOCS-1]、白介素1受体相关激酶-M[IRAK-M]和IL-10)的诱导不能解释TLR反应减弱的原因。由于CSE中含有丰富的自由基物种，GSH恢复了功能，利用免疫细胞化学筛选了一组氧化/亚硝化应激标志物。该小组确定蛋白质的羰化作用是CSE诱导的主要标志。Oxy印迹分析证实，CSE以剂量依赖和时间依赖的方式有效地向许多蛋白质引入了羰基，这与肿瘤坏死因子-α的表达呈负相关。伪足的形成没有被阻止，但这些膜延伸被严重的羰化，并且初级肺泡巨噬细胞也被作为羰化的靶点。香烟烟雾中的氧化剂推动一种快速、持久和全球性的蛋白质羰化，这可能是疾病中免疫力改变的常见途径。
Oxidants in cigarette smoke inhibit pathogen recognition receptor function and phagocytosis, but the molecular basis of this inhibition remains obscure. We sought to identify the inhibitory mechanisms that impair alveolar macrophage function. Balb/c mice were acutely exposed to four cigarettes for 4 hours before treatment with intranasal LPS (1 mu g). The mice exhibited significantly reduced airway neutrophilia and expression of TNF-alpha. Balb/c-derived MH-S alveolar macrophage cells exposed to cigarette smoke extract (CSE) displayed a similar inhibitory response to stimulation with LPS. The induction of inflammatory genes by recombinant (r) TNF-alpha(100 ng/ml) was also impaired by CSE. Because both pathways converge on NF-kappa B, the degradation of I kappa B alpha and the phosphorylation of p65 were assessed and shown to be blunted by CSE. CSE also blocked the activity of activator protein-1 (AP-1) by inhibiting p38 mitogen activated protein kinase (MAPK) in a reduced glutathione (GSH)-reversible manner. The induction of specific Toll-like receptor (TLR)-negative regulators (suppressor of cytokine signaling-1 [SOCS-1], interleukin-1 receptor associated kinase-M [IRAK-M], and IL-10) did not account for the impaired responses of TLRs. As free radical species are abundant in CSE and GSH restored function, a panel of oxidative/nitrosative stress markers was screened using immunocytochemistry. The panel identified protein carbonylation as the major CSE-inducible marker. Oxyblot analysis confirmed that CSE potently introduced carbonyl groups to many proteins in a dose-dependent and time-dependent manner that inversely correlated with the expression of TNF-alpha. The formation of pseudopodia was not prevented, but these membrane extensions were heavily carbonylated, and primary alveolar macrophages were also targeted for carbonylation. Oxidants in cigarette smoke drive a rapid, persistent, and global protein carbonylation that may represent a common pathway to altered immunity in disease.