中文摘要：

目的 探讨羟基红花黄色素A（HSYA）对低氧条件下犬血管内皮细胞（VEC）血管内皮生长因子（VEGF）相关信号传导通路的影响。方法 在低氧条件下以ELISA法观察VEGF抗体及其两种酪氨酸受体（Fit-1和KDR）的抗体对HSYA促VEC增殖作用及分泌VEGF水平的影响；生物分子相互作用分析法检测HSYA与VEGF、Fit-1和KDR的相互作用；硝酸还原酶法测定VEC培养液中NO、NOS的量。结果10μg／mLVEGF、Fit-1和KDR的抗体均能明显抑制HSYA的促VEC分泌VEGF作用；生物分子相互作用分析结果证实，HSYA与VEGF、Fit-1及KDR均无明显结合；HSYA在1mmol／L浓度下能够明显促进低氧条件下VEC培养液中的NO水平，而对NOS水平没有明显影响。结论在低氧条件下，HSYA促VEC增殖的作用与VEGF及其相关信号传导通路有关，但HSYA与VEGF及其受体无明显结合，提示可能存在与VEGF及其相关信号传导通路相关的HSYA作用靶点。

英文摘要：

Objective To study the effect of hydroxysafflor yellow A （HSYA） on vascular endothelial growth factor （VEGF） pathway in hypoxic canine aortic endothelial cell （VEC）. Methods Using several antibodies, the involvement of VEGF, faro-like tyrosine receptor （Flt-1） and kinase insert domaincontaining receptor （KDR） in HSYA-induced VEC growth was determined by MTT assay and ELISA for VEGF secretion under hypoxic condition. Furthermore, the interactions of HSYA with VEGF, Fh-1 or KDR were studied by Biomolecular Interaction Analysis （BIA）. The nitric oxide （NO） concentration and endothelial nitric oxide synthetase （eNOS） activity were determined by nitrate reductase assay. Results Antibodies of Fh-1, KDR or VEGF significantly blocked the proliferative effect of HSYA. Also, antibodies of Flt-1 and VEGF attenuated the promotion of HSYA on VEGF secretion. BIA Results revealed that HSYA had no interactions with VEGF, Flt-1, and KDR. The treatment of HSYA 1 mmol/L in hypoxic culture resulted in elevated NO level and intact NOS level in the endothelial cell culture media. Conclusion The proliferative effect of HSYA on VEC is probably mediated by VEGF pathway. However, there is no direct interaction between HSYA and VEGF or VEGF receptors, which indicates other HSYA targets in VEGF pathway.