百里醌对内皮祖细胞的抑制作用

目的探讨百里醌对人脐血来源的内皮祖细胞血管生成的抑制作用及其可能机制。方法采用贴壁选择法培养人脐血内皮祖细胞(EPCs),DiI-ac-LDL吞噬试验及VEGFR-2、Ⅷ因子和CD34细胞免疫组化证实细胞属性;百里醌作用EPCs后,CCK-8法检测细胞增殖;Transwell小室实验测定EPCs体外侵袭能力;小管形成实验检测EPCs体外小管形成能力;Western blotting检测EPCs中MMP-2和MMP-9蛋白表达。结果体外成功培养出人脐血内皮祖细胞,百里醌可显著抑制体外EPCs增殖,IC50=51.2nmol/L;百里醌可抑制体外EPCs侵袭和小管形成,呈浓度依赖性;百里醌可显著下调EPCs中MMP-2和MMP-9的表达。结论百里醌可能通过抑制EPCs中MMP-2和MMP-9的表达从而抑制EPCs参与的血管生长,从而有望作为有效的血管生成抑制药物。

Inhibitory Effect of Thymoquinone on Endothelial Progenitor Cells

Objective To investigate the anti-angiogenic effect of thymoquinone on the endothelial progenitor cells.Methods The endothelial progenitor cells(EPCs) derived from human umbilical cord blood were cultured in vitro through adhesion selection and were differentiated into endothelial cells under the induction of special cytokines.The endothelial cell lineage was confirmed by DiI-ac-LDL up-taking and immunocytochemistry of VEGFR-2,Ⅷ factor and CD34.After EPCs was treated with thymoquinone,the cellular proliferation was detected by cell counting kit-8(CCK-8) assay.The effect of thymoquinone on the invasion of EPCs was examined by using matrigel counting.A tube formation assay was performed after EPCs were cultured with thymoquinone.Western blot was used to detect the protein expression of MMP-2 and MMP-9 in EPCs.Results The EPCs derived from human umbilical cord blood were cultured in vitro successfully.The proliferation of EPCs was inhibited significantly by thymoquinone with IC50 being 51.2nmol/L.Thymoquinone significantly suppressed the invasion of EPCs.Furthermore,incubation of EPCs with thymoquinone decreased EPCs′ tube formation capacity in a concentration-dependent manner.Western bolt assay indicated that thymoquinone down-regulates the expression of MMP-2 and MMP-9 proteins in EPCs.Conclusion Thymoquinone exerts anti-angiogenic activity in EPCs,which may be related to down-regulation of MMP-2 and MMP-9 protein,could be developed as an effective anti-angiogenic drug.