重组人血管内皮细胞因子VEGF183刺激HUVEC体外增殖

Recombinant VEGF183 Protein Promotes the Proliferation of HUVEC in vitro

VEGF183为VEGF-A基因一个拼接异构体,因其蛋白序列与VEGF189高度同源,导致其功能研究被忽视,所以目前VEGF183功能仍需进一步研究。本研究通过毕赤酵母表达并用琼脂糖肝素亲和层析方法获得重组人VEGF183蛋白,MTT法研究VEGF183蛋白在体外对HUVEC增殖刺激能力,并分别与VEGF165、VEGF189的相应活性作对比。Transwell与划痕实验分别研究其对HUVEC的迁移能力,Miles assay研究其对大鼠血管通透能力,并与VEGF189的相应活性作对比。结果发现,VEGF183仍保留对HUVEC增殖和迁移的刺激能力,但弱于VEGF165,强于VEGF189。此外,VEGF183拥有强于VEGF189的刺激血管通透能力。本研究结果提示VEGF183可能为实现血管生成过程中精确调控的一个异构体。

VEGF183 is one of important VEGF isoforms,albeit it was discovered two decades ago,due to highly structural similarity with VEGF189,VEGF183 was often neglected to be investigated compared with that of VEGF189,so its function is still need to be elucidated.In this study,we expressed VEGF183 protein via pichia yeast expression and purified with HiTrap heparin column affinity chromatography.Further we tested the effect of VEGF183 on the HUVEC proliferation by MTT,HUVEC migration by Transwell and capability to rat vascular permeability by miles assay,at the same time,we compared it with those of VEGF165 or VEGF189.The results showed that VEGF183 could stimulate the proliferation,migration of HUVEC,and could also promote the vascular permeability.However,the above-mentioned capabilities of VEGF 183 are stronger than those of VEGF189,but weaker than those of VEGF165.We deduced that VEGF183 might play an elaborate role during the process angiogenesis.