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(Flt-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、Flt-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、Flt-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、Flt-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。

ObjectiveTo study the effect of hydroxysafflor yellow A (HSYA ) on vascu larendo thelial growth factor (VEGF) pathway in hypoxic canine aortic endo thelial cell(VEC). MethodsUsing several antibodies,the involvement of VEGF, fam-like tyrosine receptor (Flt21) and kinase in sert domaincon taining receptor (KDR) in HSYA-induced VEC growth was determined by MTT assay and ELISA for VEGF secretion under hypoxic condition. Furthermore, the in teractions of HSYA with VEGF, Flt21 or KDR were studied by Biomolecular In teraction A nalysis (BIA ). Then it ricoxide (NO ) concent ration and en-do thelialn it ricoxide synthetase (eNOS) activity were determined by nitrate reductase assay. ResultsAntibodies of Flt21, KDR or VEGF sign ificantly blocked the proliferative effect of HSYA. Also, antibodies of Flt21 and VEGF at tenuated the promotion of HSYA on VEGF secretion. BIA Results revealed that HSYA had no in teractions with VEGF, Flt21, and KDR. The treatment of HSYA1 mmolöL in hypoxic culture resulted in elevated NO level and intact NO S level in the endo thelial cell culture media. ConclusionThe proliferative effect of HSYA on VEC is probab lymediated by VEGF pathway. However, there is no direct in teract on between HSYA and VEGF or VEGF receptors, which indicates other HSYA targets in VEGF pathway.

国家自然科学基金资助项目(30370720);国家“863”计划(2004AA2Z3815)