血府逐瘀胶囊调控EphB4/EphrinB2促血管适度生长的实验研究

Moderate Angiogenic Effect of Xuefu Zhuyu Capsule by Regulating EphB4/EphrinB2

目的研究血府逐瘀胶囊适度促血管新生的作用及其对EphB4/EphrinB2表达的影响。方法通过检测血府逐瘀胶囊含药血清对人微血管内皮细胞(human microvascular endothelial cell line 1,HMEC-1)活性、细胞周期、迁移、黏附和体外成血管的影响,明确药物具有适度促血管新生的作用,并运用实时定量PCR、Western blot技术检测药物对EphB4/EphrinB2表达的影响。结果血府逐瘀含药血清对内皮细胞活性及S期细胞比例均无影响。细胞迁移方面,1.25%含药血清作用24、48、72 h均显著增加细胞迁移能力;但2.50%含药血清在最初24 h抑制细胞迁移,随后显著促进迁移,与5.00%含药血清的影响趋势正好相反。细胞黏附方面,1.25%含药血清作用72 h时显著减少黏附细胞数;2.50%药物血清在前两个时间段增加细胞黏附数,随后在72 h抑制细胞黏附;5%含药血清在24 h表现出抑制作用,而在48 h转为促进,随后作用消失。成血管方面,仅2.50%含药血清表现出促进作用,而5.00%含药血清作用48 h和72 h均出现抑制现象。2.50%含药血清组的实时定量PCR和Western blot结果显示,药物作用12 h上调EphB4的表达,作用24 h下调EphB4表达的同时上调EphrinB2的表达。结论仅2.50%含药血清作用48 h对HMEC-1的迁移、黏附和体外成血管均有促进作用,为最佳促血管生长条件,提示血府逐瘀胶囊促血管新生作用具有一定的条件限制,而药物调控EphB4/EphrinB2的表达是其重要因素之一。

Objective To observe moderate angiogenic effect of Xuefu Zhuyu Capsule（ XFZYC）on human microvascular endothelial cell line 1（ HMEC-1）, and its regulation effect on expression of EphB4 / EphrinB2.Methods The moderate angiogenic effect of XFZYC was clarified by detecting XFZYC containing serum on cell viability,cell cycle,migration,adhesion and in vitro angiogenesis.Its effects on expressions of EphB4 / EphrinB2 were detected by Real-time quantitative PCR and W estern blot.Results XFZYC containing serum（ XFZYC-CS） had no effect on the cell viability or cell ratio in phase S endothelial cells.Cell migration was significantly improved by 1.2 5 % XFZYC-CS after 2 4,4 8,and 7 2 h of action2.5 0 % XFZYC-CS inhibited cell migration at the primary 2 4 h,but it significantly promoted cell migration at 4 8 and 7 2 h afterwards.It showed just an opposite tendency to 5.0 0 % XFZYC-CS.Cellular adhesion number was significantly reduced by 1.2 5 % XFZYC-CS at 7 2 h.Cellular adhesion number was significantly increased by 2.5 0 % XFZYC-CS at 2 4 and 4 8 h,but inhibited at 7 2 h 5.0 0 % XFZYC-CS showed inhibition at 2 4 h,but turned to promotion,and disappeared afterwards.In vessel formation aspect,only 2.5 0 % XFZYC-CS showed vessel formation promotion 5.0 0 % XFZYC-CS showed inhibition on vessel formation at 4 8 and 7 2 h.Results of Real-time quantitative PCR and W estern blot in 2.5 0 %XFZYC-CS showed EphB4 expression was up-regulated at 1 2 h; EphB4 expression was down-regulatedwhile EphrinB2 expression was up-regulated at 2 4 h.Conclusions Only 2.5 0 % XFZYC-CS at 4 8 h had promotion of migration,adhesion,andin vitro angiogenesis of H MEC-1,which was the optimal condition for vessel growth.These results suggested XFZYC promoted angiogenesis in certain conditional limitations.But it regulated the expression of EphB4 / EphrinB2,which might be one of important factors.