CYP2J subfamily P450s in the lung: expression, localization, and potential functional significance.
CYP2J subfamily P450s in the lung: expression, localization, and potential functional significance.
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发表时间：
1996-11
影响因子：
3.6
通讯作者：
D. Zeldin;J. Foley;Jixiang Ma;J. Boyle;J. Boyle;Jorge M. Pascual;Jorge M. Pascual;C. Moomaw;K. Tomer;C. Steenbergen;Shu Wu
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文献类型：
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作者：
D. Zeldin;J. Foley;Jixiang Ma;J. Boyle;J. Boyle;Jorge M. Pascual;Jorge M. Pascual;C. Moomaw;K. Tomer;C. Steenbergen;Shu Wu
细胞色素P450(P450)单加氧酶催化花生四烯酸环氧化生成环氧二十碳三烯酸，调节支气管平滑肌张力和呼吸道跨上皮离子转运。我们最近描述了一种新的人类P450花生四烯酸环氧合酶(CYP2J2)及其相应的大鼠同源物(CYP2J3)。用CYP2J基因探针对肺组织RNA进行Northern分析，结果表明，肺组织中表达了细胞色素P450 2J2和细胞色素P450 2J3mRNAs。用抗重组人CYP2J2的多克隆抗体免疫印迹人和大鼠肺微粒体组分，发现一条单一的56 kDa条带，证实肺组织中有丰富的CYP2J2和CYP2J3蛋白表达。对福尔马林固定、石蜡包埋的人和大鼠肺切片进行抗人CYP2J2抗体免疫组织化学分析和抗生物素/过氧化物酶检测，结果表明，CYP2J蛋白主要在纤毛上皮细胞中表达。无纤毛的呼吸道上皮细胞、支气管和肺血管平滑肌细胞、肺血管内皮细胞和肺泡巨噬细胞也有较强的染色，而肺泡上皮细胞的染色较弱。用气相色谱/质谱仪在人和大鼠肺组织中检测到内源性环氧二十碳三烯酸，为花生四烯酸在人和大鼠肺内P450的代谢提供了直接证据。根据这些数据，我们得出结论：CYP2J2和CYP2J3是丰富的肺花生四烯酸环氧合酶，其产物是环氧二十碳三烯酸，是人和大鼠肺的内源性成分。除了对气道平滑肌张力和跨上皮电解质转运的已知影响外，CYP2J蛋白在血管平滑肌和内皮的定位表明，环氧二十碳三烯酸可能也参与了肺血管张力的调节。
Cytochrome P450 (P450) monooxygenases catalyze the epoxidation of arachidonic acid to form epoxyeicosatrienoic acids, which modulate bronchial smooth muscle tone and airway transepithelial ion transport. We recently described a new human P450 arachidonic acid epoxygenase (CYP2J2) and the corresponding rat homologue (CYP2J3). Northern analysis of lung RNA using CYP2J cDNA probes demonstrated that CYP2J2 and CYP2J3 mRNAs were expressed in the lung. Immunoblotting of microsomal fractions prepared from human and rat lungs using a polyclonal antibody raised against recombinant human CYP2J2 revealed a single 56-kDa band confirming abundant pulmonary CYP2J2 and CYP2J3 protein expression. Immunohistochemical analysis of formalin-fixed paraffin-embedded human and rat lung sections using the anti-human CYP2J2 IgG and avidin/biotin/peroxidase detection showed that CYP2J proteins were primarily expressed in ciliated epithelial cells lining the airway. Prominent staining was also noted in nonciliated airway epithelial cells, bronchial and pulmonary vascular smooth muscle cells, pulmonary vascular endothelium, and alveolar macrophages, whereas less intense staining was noted in alveolar epithelial cells. Endogenous epoxyeicosatrienoic acids were detected in both human and rat lung using gas chromatography/mass spectrometry, thus providing direct evidence for the in vivo human and rat pulmonary P450 metabolism of arachidonic acid. Based on these data, we conclude that CYP2J2 and CYP2J3 are abundant pulmonary arachidonic acid epoxygenases and that CYP2J products, the epoxyeicosatrienoic acids, are endogenous constituents of human and rat lung. In addition to known effects on airway smooth muscle tone and transepithelial electrolyte transport, the localization of CYP2J proteins to vascular smooth muscle and endothelium suggests that epoxyeicosatrienoic acids may also be involved in the modulation of pulmonary vascular tone.