Oxidant stress stimulates mucin secretion and PLC in airway epithelium via a nitric oxide-dependent mechanism.
Oxidant stress stimulates mucin secretion and PLC in airway epithelium via a nitric oxide-dependent mechanism.
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氧化应激通过一氧化氮依赖性机制刺激气道上皮中的粘蛋白分泌和 PLC。
DOI：
10.1152/ajplung.1996.271.5.l854
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Adler,KB
中科院分区：
文献类型：
--
作者：
Wright,DT;Fischer,BM;Li,C;Rochelle,LG;Akley,NJ;Adler,KB
活性氧（ROS）参与了多种呼吸系统疾病的发病机制。我们研究了原代培养的豚鼠气管上皮细胞（GPTE）的ROS诱导粘蛋白分泌的机制，以及GPTE细胞和来源于人支气管上皮的病毒转化细胞系（BEAS-2B）中ROS诱导的第二信使产生酶磷脂酶C（PLC）的激活。粘蛋白分泌测定单克隆抗体为基础的酶联免疫吸附试验，和PLC激活进行了评估，阴离子交换色谱。在嘌呤（500 μ M）存在下，由黄嘌呤氧化酶（XO，500 μ M）酶促产生的ROS增强GPTE细胞的粘蛋白释放，并激活GPTE和BEAS细胞中的PLC。粘蛋白的过度分泌和PLC对嘌呤+ XO的反应似乎是通过依赖于内源性产生的一氧化氮和可能的细胞内产生的氧化剂的细胞内途径发生的。这两种反应都可以被阻断或减弱的细胞与NG-单甲基-L-精氨酸，一氧化氮合酶的抑制剂，或与二甲基硫脲，一种化合物，可以与各种细胞内氧化剂物种的预孵育。化学产生的活性氮物质也刺激粘蛋白的分泌，并通过依赖于（至少部分地）细胞内氧化剂介导的过程的机制激活PLC。结果表明，细胞内产生的自由基种类的氮和氧可能是重要的调制器的气道上皮细胞对外部氧化应激的反应。
Reactive oxygen species (ROS) have been implicated in the pathogenesis of a wide variety of respiratory diseases. We investigated mechanisms of ROS-induced mucin secretion by guinea pig tracheal epithelial (GPTE) cells in primary culture, and ROS-induced activation of the second messenger-producing enzyme phospholipase C (PLC), in GPTE cells and in a virally transformed cell line (BEAS-2B) derived from human bronchial epithelium. Mucin secretion was measured by a monoclonal antibody-based enzyme-linked immunosorbent assay, and PLC activation was assessed by anion exchange chromatography. ROS generated enzymatically by xanthine oxidase (XO, 500 microM) in the presence of purine (500 microM) enhanced release of mucin by GPTE cells and activated PLC in GPTE and BEAS cells. Hypersecretion of mucin and activation of PLC in response to purine + XO appeared to occur via an intracellular pathway(s) dependent on endogenously produced nitric oxide and possibly intracellularly generated oxidants. Both responses could be blocked or attenuated by preincubation of the cells with NG-monomethyl-L-arginine, an inhibitor of the enzyme nitric oxide synthase, or with dimethylthiourea, a compound that can react with a variety of intracellular oxidant species. Reactive nitrogen species generated chemically also stimulated secretion of mucin and activated PLC via a mechanism dependent (at least in part) on intracellular oxidant-mediated process(es). The results suggest that intracellularly generated radical species of nitrogen and oxygen may be important modulators of the response of airway epithelial cells to external oxidant stress.