细胞色素酶CYP2C19介导的沙利度胺体外抗血管生成作用研究

本研究探讨人肝细胞微粒体代谢系统对治疗多发性骨髓瘤的沙利度胺体外抗血管生成的影响及细胞色素酶CYP2C19在其中的作用。采用沙利度胺原药或与人肝细胞微粒体在体外共孵育后用MTY法检测人脐带静脉内皮细胞（human umbilical cord vein endothelial cells,hUCVEC）增殖活力,用流式细胞术测定hUVCEC细胞周期和细胞凋亡,以改良的Boyden小室法检测hUCVEC细胞迁移力,以体外小管形成实验检测hUCVEC分化。结果表明：沙利度胺原药对hUCVEC活力无明显抑制作用,细胞凋亡比例也无明显增加,轻度影响细胞迁移,无抗小管形成作用;当与人肝细胞微粒体共孵育后hUCVEC增殖活力明显受抑。100μg／ml沙利度胺与肝细胞微粒体共孵育后hUCVEC增殖活力的抑制率达（11．7±3．9）％,凋亡细胞增加达27．2％,明显下调细胞迁移力并抑制体外小管形成。在共孵育体系中加入CYP2C19特异性抑制剂奥美拉唑,可减弱沙利度胺抑制hUCVEC增殖活力和诱导凋亡的作用,减低细胞迁移力和部分逆转抗小管形成的作用。结论：沙利度胺的体外抗血管生成作用依赖于人细胞微粒体的作用,细胞色素酶系中的CYP2C19可能参与了这一过程。

Antiangiogenic Activity of Thalidomide In Vitro Mediated by Cytochrome CYP2 C19

This study was aimed to investigate the effect of metabolic system in human hepatic cell rnicrosome on antiangiogenic in vitro activity of thalidomide used in treating multiple myeloma and to explore the role of cytochrome CYP2C19. Human umbilical cord vein endothelial cells （hUCVECs） were treated with thalidomide alone or thalidomide co-incubated with human hepatic cell microsome. Cell proliferation ability was assessed by MTT assay, cell cycle analysis and detection of apoptosis were carded out by flow cytomety （ FCM）, migration activity of hUCVECs was deter- mined by modified Boyden chamber and differentiation of hUCVECs was assayed by tube formation test. The results showed that thalidomide alone had no obvious direct effect on hUCVEC viability or apoptosis, mild effect on cell migration and no effect on tube formation. However, when co-incubated with human hepatic cell microsome, thalidomide significantly inhibited the hUCVECs viability. At 100 μg/ml, thalidomide co-incubated with human hepatic cell microsome, the proliferation ability of hUCVECs decreased by （ 11.7±3.9 ） % , apoptosis cells increased by 27.2%, the cell migration was down-regulated significantly, and the tube formation was obviously inhibited. When omeprazole, a specific inhibitor of cytochrome CYP2C19, was added into the co-incubation mixture, the effects of thalidomide on cell proliferation ability, apoptosis, migration and tube formation decreased. It is concluded that effect of human hepatic cell microsome is required for thalidomide＇s antiangiogenic activity in vitro and cytochrome CYP2C19 may be involved in the antiangiogenic effect of thalidomide.