Identification and localization of five CYP2Cs in murine extrahepatic tissues and their metabolism of arachidonic acid to regio- and stereoselective products.
Identification and localization of five CYP2Cs in murine extrahepatic tissues and their metabolism of arachidonic acid to regio- and stereoselective products.
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发表时间：
2001-10
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通讯作者：
C. Tsao;S. Coulter;Anna Chien;Gang Luo;N. Clayton;R. Maronpot;J. Goldstein;D. Zeldin
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作者：
C. Tsao;S. Coulter;Anna Chien;Gang Luo;N. Clayton;R. Maronpot;J. Goldstein;D. Zeldin
CYP 2C亚家族已在人体中就临床重要药物的代谢进行了广泛研究，并已在这些酶中鉴定出多态性。在本研究中，使用小鼠模型来确定可能的生理功能和肝外分布的CYP 2C。本文采用逆转录-聚合酶链反应（RT-PCR）、免疫印迹和免疫组化等方法，证实小鼠CYP 2C广泛分布于肝外组织，并定位于心肌、肺Clara细胞和纤毛细胞、肾集合管、雌性肾上腺X区、生殖器官、白色血细胞和眼（视神经、视杆细胞和视锥细胞）。产物的RT-PCR、亚克隆和测序表明，每种CYP 2C具有独特的组织分布。四个cDNA片段代表潜在的新的CYP 2C被确定，每个都有自己的器官特异性表达模式。使用细菌cDNA表达系统，我们发现，重组蛋白的五个全长小鼠CYP 2C代谢花生四烯酸不同的区域和立体特异性产品，包括epoxyeicosatrienoic酸和hydroxyyeicosatrienoic酸。已报道花生四烯酸的区域特异性和立体特异性代谢物影响重要的生理功能，如炎症、中性粒细胞活化、离子转运、细胞增殖和血管张力。我们的研究结果表明，CYP 2C酶在心肌，主动脉，肾，肺，肾上腺，眼睛和生殖器官的存在下，可以调节这些组织中重要的生理和/或病理过程。
The CYP2C subfamily has been extensively studied in humans with respect to the metabolism of clinically important drugs, and polymorphisms have been identified in these enzymes. In the present study, a murine model was used to determine the possible physiological functions and extrahepatic distribution of CYP2Cs. Using the reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immununohistochemistry, this report demonstrates that the mouse CYP2Cs are extensively distributed in extrahepatic tissues and localized to heart muscle, lung Clara and ciliated cells, kidney collecting ducts, the X-zone of female adrenals, reproductive organs, white blood cells, and eyes (in the optic nerve, rods, and cones). RT-PCR, subcloning, and sequencing of the products indicate that each CYP2C has a unique tissue distribution. Four cDNA fragments representing potentially new CYP2Cs were identified, each with its own organ-specific pattern of expression. Using a bacterial cDNA expression system, we found that recombinant proteins for each of the five full-length murine CYP2Cs metabolize arachidonic acid to different regio- and stereospecific products, including epoxyeicosatrienoic acids and hydroxyeicosatetraenoic acids. Regio- and stereospecific metabolites of arachidonic acid have been reported to affect important physiological functions such as inflammation, neutrophil activation, ion transport, cellular proliferation, and vascular tone. Our results suggest that the presence of CYP2C enzymes in heart muscle, aorta, kidney, lung, adrenals, eyes, and reproductive organs could regulate important physiological and/or pathological processes in these tissues.