羟基红花黄色素A抑制异常增殖血管内皮细胞的机制研究

Hydroxysafflor yellow A inhibited abnormal proliferation of vascular endothelial cells

目的研究羟基红花黄色素A(HSYA)抑制异常增殖血管内皮细胞的作用机理。方法建立人结肠腺癌细胞LS180上清液和人脐静脉内皮细胞ECV304体外共培养模型,通过实时荧光定量PCR检测ECV304细胞中血管内皮生长因子(VEGF)及其受体(KDR)、碱性成纤维细胞生长因子(b FGF)及其受体(b FGFR)、HSPG2、p53、c-myc mRNA水平的表达;ELISA法检测细胞培养上清VEGF、VEGFR(KDR)、b FGF、b FGFR蛋白的表达。结果在共培养细胞模型中,0.66、0.33 mg/L浓度的HSYA对血管生成促进因子VEGF及其受体KDR、b FGF及其受体b FGFR和癌基因c-myc及与细胞增殖相关蛋白多糖HSPG2的mRNA表达具有抑制作用;对抑癌基因p53 mRNA的表达具有促进作用。0.66、0.33 mg/L浓度的HSYA对血管生成促进因子VEGF及其受体KDR、b FGF及其受体b FGFR的蛋白表达具有抑制作用。结论 HSYA抑制新生血管的生成、抑制共培养模型中血管内皮细胞的异常增殖,其可能的作用机理是通过调控VEGF及其受体、b FGF及其受体、HSPG2、c-myc和野生型p53的表达发挥抗肿瘤作用,揭示HSYA可能是潜在的肿瘤血管生成抑制剂。

Objective To study the inhibitory mechanism of hydroxysafflor yellow A（ HSYA） on abnormal proliferation of vascular endothelial cells. Methods Co-cultured model in vitro was established,with supernatant fraction of LS180 human colon adenocarcinoma cells tumor cells and ECV304 human umbilical vein endothelial cells. Then mRNA expressions of vascular endothelial growth factors（ VEGF）and VEGF receptors（ KDR）,basic fibroblast growth factor（ b FGF） and b FGF receptors（ b FGFR）,proteoglycan related to cell proliferation（ HSPG2）,p53 and c-myc were detected using real-time fluorescence quantitative PCR; protein expressions of VEFG,KDR,b FGF and b FGFR in supernatant fraction were measured by ELISA. Results In the cell co-cultured model,the HSYA concentration of 0. 66 and0. 33 mg/L inhibited mRNA and protein expressions of VEGF,KDR,b FGF and b FGFR,inhibited mRNA expressions of c-myc and HSPG2,while upregulated p53 mRNA expression. Conclusion HSYA inhibited angiogenesis and abnormal proliferation of vascular endothelial cells in co-cultured model by regulating expressions of VEGF,VEGFR,b FGF,b FGFR,HSPG2,c-myc,wild type p53. HSYA may be a potential tumor angiogenesis inhibitor.