辛二酰苯胺氧肟酸对血管生成的抑制作用及其机制

Inhibitory effect of suberoylanilide hydroxamic acid on angiogenesis and its molecular mechanism

目的研究辛二酰苯胺氧肟酸(SAHA)对人脐静脉内皮细胞(HUVEC)生物学特性的影响及对血管生成的作用。方法 CCK-8法检测SAHA对HUVEC增殖能力的影响。TranswellTM小室法、基质胶体外成管实验、Western blot法、基质胶体内血管生成实验分别观察15μmol/L SAHA处理HUVEC 48 h后HUVEC迁移能力的变化;HUVEC在基质胶上形成管腔样结构的能力;HUVEC细胞中血管内皮生长因子(VEGF)信号通路相关蛋白表达的变化以及小鼠体内基质胶栓中新生血管的数量。结果 SAHA能够抑制HUVEC的增殖,抑制作用呈剂量、时间依赖性;与对照组相比较,SAHA处理组迁移过膜的HUVEC细胞数明显降低;HUVEC形成的管腔样结构SAHA处理组明显少于对照组;SAHA抑制HUVEC中VEGF相关信号通路;SAHA处理后小鼠体内的基质胶栓中新生的血管数明显少于对照组。结论 SAHA能够抑制HUVEC增殖及体内外血管形成能力,并且其可能机制是SAHA阻断了VEGF相关的信号通路。

Objective To study the effect of histone deacetylase inhibitor （HDACi）, suberoylanilide hydroxamic acid （SAHA） on the angiogenesis and the biological characteristics of human umbilical vein endothelial cells （HUVECs）. Methods The proliferation of HUVECs affected by SAHA was investigated by CCK-8. After treated with 15 μmol/L SAHA for 48 hours, HUVECs underwent the TranswellTM assay to observe the migration, the Matrigel angiogenesis assay in vitro to test the ability of tube formation, Western blotting to evaluate the change of vascular endothelial growth factor （VEGF）-induced signaling pathway, and the Matrigel-based tube formation assay in vivo to observe the new formed vessel in matrigel plug, thereby evaluating the effect of SAHA on the angiogenesis in mice. Results Compared with the control group, SAHA significantly suppressed HUVECs proliferation in a dose-dependent and time-dependent manner. The number of migrated HUVECs was significantly reduced after the treatment of 1,5 μmol/L SAHA for 48 hours. The Matrigel angiogenesis in vitro suggested that the capillary-like tube formation in the SAHA treated group was less than that in the control group. Western blotting demonstrated that SAHA inhibited the VEGF-induced signaling pathway. The number of new formed vessels in the Matrigel plug in the SAHA treated group was less than that in the control group. Conclusion SAHA suppresses the proliferation of HUVECs and the angiogenensis in vivo and in vitro. The potential mechanism of the inhibition may be SAHA blockage of the VEGF-induced signaling pathway.