麦考酚酸对flk-GFP斑马鱼模型抗血管生成的作用机制研究

Study on effective mechanism of mycophenolic acid on antiangiogenesis in flk-GFP zebrafish

目的从已上市药物中筛选出具有抑制血管生成作用的小分子化合物麦考酚酸(免疫抑制剂)并对其抗血管生成作用机制进行研究。方法用flk-GFP转基因斑马鱼作为观察血管生成的动物模型,从含有大部分已上市药物的1120种化合物库(Prestwick Chemical,Inc.)中,筛选出具有典型抑制血管生成的药物麦考酚酸(MPA);对其抑制血管生成的作用进行量效分析,通过目标基因敲出技术和微注射方法,研究flk-GFP转基因斑马鱼胚胎注射次黄嘌呤核苷磷酸脱氢酶(IMPDH)的吗啉反义寡核苷酸(MO)3种亚型的血管生成抑制作用。结果麦考酚酸抑制血管生成的作用与药物浓度成正相关,且有明显的血管生成阻断作用,提示IMPDH2 MO能显著抑制斑马鱼的血管生成(n>200,表型比例>50%)。结论麦考酚酸通过抑制IMPDH2从而产生血管生成抑制作用。

Objective To identify small molecules with potential of antiangiogenesis （mycophenolic acid, MPA） from approved drugs and do further study for its mechanism of action. Methods The flk - GFP zebrafish serves as a model for rapid observation of angiogenesis in living embryos. We screened 1 120 compounds library and revealed that MPA exhibited effect of inhibition of blood vessels but had no other detrimental effects on general embryonic development. The mechanism of MPA action was further analyzed by microangiographic studies and 3 subtypes IMP-DH, the possible target of MPA, morpholino oligos （ MO ） - knock. Resuits The inhibition of angiogenesis caused by MPA is dose - dependent. The circulation of these embryos was also blocked as revealed by microangiographic analysis, suggesting potent inhibition of angiogenesis. It was showed that IMPDH 2 is mainly expressed in the ventral regions of the developing trunk and IMPDH2 antisense morpholino can inhibit angiogenesis of zebrafish （ n 〉 200, 〉 50% showed phenotype）. Conclusion MPA can induce antiangiogenesis by inhibiting IMPDH2.