摘要
目的:评价活血化瘀方(SE)的促血管新生作用,并探讨其可能的作用机制。方法:选择转基因斑马鱼幼鱼及内皮细胞系EA.hy926细胞株为模型,按空白对照组、模型组、活血化瘀方不同剂量组进行分组和给药干预。在荧光显微镜下观察正常斑马鱼胚胎培养液中空白对照组及活血化瘀方组肠下静脉血管(SIVs)的生长状况;用VEGF受体抑制剂(VRI)诱导斑马鱼血管损伤,观察各组斑马鱼节间血管(ISVs)生长情况;采用VRI抑制内皮细胞系EA.hy926增殖模型,用MTT法评价活血化瘀方各组对细胞增殖的促进作用;用Real-time PCR法,检测各组斑马鱼体内血管内皮生长因子受体(VEGFR)基因(flt1、kdr、kdrl)的表达情况。结果:在正常斑马鱼模型上,活血化瘀方可明显促进斑马鱼SIVs出芽;在VRI诱导的斑马鱼血管损伤模型上,活血化瘀方各剂量组能够明显抑制斑马鱼ISVs的损伤,其血管生长指数与模型组比较差异均有统计学意义(P<0.001);在EA.hy926细胞模型上,活血化瘀方各剂量组可明显促进内皮细胞的增殖。Real-time PCR结果表明,模型组VRI显著抑制了斑马鱼体内VEGF受体基因flt1、kdr、kdrl的表达,而活血化瘀方各剂量组的表达量均明显增高,并呈现一定的剂量依赖关系。结论:活血化瘀方在内皮细胞模型、正常斑马鱼模型和VRI诱导的斑马鱼血管损伤模型上均具有促血管新生作用,其机制可能与其上调VEGF受体flt1、kdr、kdrl基因的表达有关。
Objective： To evaluate the pro-angiogenic effect of SE Formula and its action mechanism study. Methods：Transgenic zebrafish embryos Tg（ fli-1a： EGFP） y1 and endothelial cell line EA. hy926 were used as experimental models in this study. All experiments contained blank control,model and different concentration SE Formula treatment groups,with intervention by drug adminstration respectively. Zebrafish morphological changes were observed under fluorescence microscope.The difference of angiogenic vessel formation（ spout） in subintestinal vessels（ SIVs） was compared between control and SE Formula treatment groups. SE Formula treatment inhibiting VRI-induced intersegmental vessels（ ISVs） loss was concerned.The proliferation effect of SF Formula was evaluated in VRI-induced EA. hy926 cell growth inhibition model by cell viability assay（ MTT）. The relative mRNA expression levels of VEGFRs including flt1,kdr and kdrl were tested by Real-time PCR after SF Formula treatment in zebrafish embryos. Results： SE Formula increased angiogenic vessel formation（ spout） in SIVs in zebrafish obviously. And SE Formula protected VRI-induced ISVs loss and the SIV index was significantly increased in SE Formula treatment groups,and there was significant difference in SIV index compared with model group（ P〈0. 001）.Moreover,SE Formula could promote proliferation effect in EA. hy926 cells. As determined by quantitative real-time PCR,the dose-dependent up-regulation of VEGFRs including kdr, kdrl and flt1 was detected in SE Formula treatment group.Conclusion： This study indicates SE Formula presents pro-angiogenic effect both in zebrafish embryos and EA. hy926 cells,and its action mechanism may relate to up-regulating the expression of VEGFRs including fltl,kdr and kdrl.
出处
《上海中医药大学学报》
CAS
2016年第1期40-44,共5页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
国家自然科学基金海外及港澳学者合作项目(81328025)
上海市自然科学基金项目(13ZR1442400)
上海市卫计委科研基金项目(20154Y0052)
上海中医药大学预算内项目(2014YSN31)
