Chronic Alcohol Ingestion Impairs Rat Alveolar Macrophage Phagocytosis via Disruption of RAGE Signaling.
Chronic Alcohol Ingestion Impairs Rat Alveolar Macrophage Phagocytosis via Disruption of RAGE Signaling.
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DOI：
10.1016/j.amjms.2017.12.013
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发表时间：
2018-05
期刊：
影响因子：
--
通讯作者：
Guidot DM
中科院分区：
文献类型：
--
作者：
Staitieh BS;Egea EE;Fan X;Amah A;Guidot DM
酒精显著损害肺的抗氧化防御和先天免疫功能，并增加基质金属蛋白酶9(MMP9)的活性。晚期糖基化终产物受体(RAGE)是一种特征明确的肺损伤标志物，可被基质金属蛋白酶-9裂解为可溶性RAGE，目前尚未在酒精性肺中检测到。我们假设慢性酒精摄入会损害肺泡巨噬细胞中通过基质金属蛋白酶-9传递的RAGE信号，从而损害先天免疫功能。原代肺泡巨噬细胞分离自对照组和酒精喂养的大鼠。采用实时定量聚合酶链式反应(qRT-PCR)、Western blotting和酶联免疫吸附试验检测RAGE的表达。通过qRT-PCR证实大鼠肺泡巨噬细胞系中基质金属蛋白酶-9核糖核酸(RNA)的沉默，并用免疫荧光(IF)法评估酒精、基质金属蛋白酶-9和RAGE之间的关系。用流式细胞仪检测细胞吞噬功能。用萝卜硫素和谷胱甘肽评估氧化应激和RAGE之间的关系。酒精喂养大鼠肺泡巨噬细胞RAGE信使RNA表达显著增加，但膜结合型RAGE蛋白表达降低。灌洗液中可溶性RAGE(SRAGE)水平升高。使用si-MMP9降低MMP9的表达可消除酒精对RAGE蛋白的影响。吞噬功能可被直接RAGE抑制，这种损害可被抗氧化剂逆转。慢性酒精摄入减少了RAGE蛋白的表达，并增加了肺泡灌洗液中的sRAGE的量，这可能是通过被基质金属蛋白酶-9切割而实现的。此外，它还会损害吞噬功能。抗氧化剂可恢复膜结合的RAGE和吞噬功能。
Alcohol significantly impairs antioxidant defenses and innate immune function in the lung and increases matrix metalloproteinase 9 (MMP-9) activity. The receptor for advanced glycation end products (RAGE) is a well-characterized marker of lung injury that is cleaved by MMP-9 into soluble RAGE and has not yet been examined in the alcoholic lung. We hypothesized that chronic alcohol ingestion would impair RAGE signaling via MMP-9 in the alveolar macrophage and thereby impair innate immune function. Primary alveolar macrophages were isolated from control-fed or alcohol-fed rats. Real-time polymerase chain reaction (qRT-PCR), Western blotting, and enzyme-linked immunosorbent assays were performed to evaluate RAGE expression. Silencing of MMP-9 ribonucleic acid (RNA) in a rat alveolar macrophage cell line was confirmed by qRT-PCR and immunofluorescence (IF) was utilized to assess the association between alcohol, MMP-9, and RAGE. Phagocytosis was assessed using flow cytometry. Sulforaphane and glutathione were used to assess the relationship between oxidative stress and RAGE. RAGE messenger RNA expression was significantly increased in the alveolar macrophages of alcohol-fed rats, but IF showed membrane-bound RAGE protein expression was decreased. Lavage fluid demonstrated increased levels of soluble RAGE (sRAGE). Decreasing MMP-9 expression using si-MMP-9 abrogated the effects of alcohol on RAGE protein. Phagocytic function was suppressed by direct RAGE and the impairment was reversed by antioxidant treatment. Chronic alcohol ingestion reduces RAGE protein expression and increases the amount of sRAGE in alveolar lavage fluid, likely via cleavage by MMP-9. In addition, it impairs phagocytic function. Antioxidants restore membrane-bound RAGE and phagocytic function.
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DOI：
10.1164/rccm.201301-0061oc
发表时间：
2013-09-15
影响因子：
24.7
作者：
Mehta, Ashish J.;Yeligar, Samantha M.;Guidot, David M.
通讯作者：
Guidot, David M.
影响因子：
4.4
作者：
Joshi, PC;Applewhite, L;Guidot, DM
通讯作者：
Guidot, DM
DOI：
10.1164/ajrccm.160.4.9811060
发表时间：
1999-10-01
影响因子：
24.7
作者：
Lois, M;Brown, LAS;Guidot, DM
通讯作者：
Guidot, DM
DOI：
10.1111/j.1530-0277.2001.tb02320.x
发表时间：
2001-07-01
影响因子：
3.2
作者：
Brown, LAS;Harris, FL;Guidot, DM
通讯作者：
Guidot, DM
影响因子：
8.8
作者：
Jabaudon, Matthieu;Futier, Emmanuel;Constantin, Jean-Michel
通讯作者：
Constantin, Jean-Michel