Objective： To evaluate the effect of Xuefu Zhuyu Capsule（血府逐瘀胶囊）-containing serum（XFZY-CS） on Eph B4/ephrin B2 and its reverse signal in human microvascular endothelial cell-1（HMEC-1）. Methods： XFZY-CS a...Objective： To evaluate the effect of Xuefu Zhuyu Capsule（血府逐瘀胶囊）-containing serum（XFZY-CS） on Eph B4/ephrin B2 and its reverse signal in human microvascular endothelial cell-1（HMEC-1）. Methods： XFZY-CS and the blank control serum were collected. HMEC-1 cells were randomly assigned to 6 groups including the concentration 1.25%, 2.5%, and 5% XFZY-CS groups and their blank serum control ones. The angiogenesis effect of XFZY-CS was tested with an in vitro tube formation assay and the best condition of pro-angiogenesis was determined. The effect of XFZY-CS on Eph B4/ephrin B2 and the reverse signal were determined by Western blot and real-time quantitative polymerase chain reaction, respectively; we also confirmed the results through activating and inhibiting the reverse signal by Eph B4/fc and pyrophosphatase/phosphodiesterase2（PP2）. Results： XFZY-CS promoted angiogenesis at the concentration of 2.5% corresponding serum after being cultured for 48 h, while inhibited angiogenesis at the concentration of 5% after culturing for 48 and 72 h. Under the 2.5% serum concentration, XFZY up-regulated the expression of Eph B4-m RNA at 12 h（P〈0.05）, and down-regulates its expression at 24 h（P〈0.01）. Protein expression of Eph B4 was apparently up-regulated at 12 h and down-regulated at 24 h. The phosphorylation of ephrin B2 increased at 9 h（P〈0.05）. In addition, 2.5% XFZY-CS played a similar role as the reverse signaling activator Eph B4/Fc ranging from 0.5 to 5 μg/m L（P〉0.05）. XFZY-CS also reduced the inhibitive effect of PP2 in limited periods. Conclusion： Eph B4/ephrin B2 was the upstream signal in the process of angiogenesis and its reverse signaling was responsible for XFZY＇s effect on promoting angiogenesis.展开更多
基金Supported by the National Natural Science Foundation of China(No.81072933 and No.81173431)National Center for Complementary and Alternative Medicine/National Institutes of Health of USA(No.2K24 AT004095)
文摘Objective： To evaluate the effect of Xuefu Zhuyu Capsule（血府逐瘀胶囊）-containing serum（XFZY-CS） on Eph B4/ephrin B2 and its reverse signal in human microvascular endothelial cell-1（HMEC-1）. Methods： XFZY-CS and the blank control serum were collected. HMEC-1 cells were randomly assigned to 6 groups including the concentration 1.25%, 2.5%, and 5% XFZY-CS groups and their blank serum control ones. The angiogenesis effect of XFZY-CS was tested with an in vitro tube formation assay and the best condition of pro-angiogenesis was determined. The effect of XFZY-CS on Eph B4/ephrin B2 and the reverse signal were determined by Western blot and real-time quantitative polymerase chain reaction, respectively; we also confirmed the results through activating and inhibiting the reverse signal by Eph B4/fc and pyrophosphatase/phosphodiesterase2（PP2）. Results： XFZY-CS promoted angiogenesis at the concentration of 2.5% corresponding serum after being cultured for 48 h, while inhibited angiogenesis at the concentration of 5% after culturing for 48 and 72 h. Under the 2.5% serum concentration, XFZY up-regulated the expression of Eph B4-m RNA at 12 h（P〈0.05）, and down-regulates its expression at 24 h（P〈0.01）. Protein expression of Eph B4 was apparently up-regulated at 12 h and down-regulated at 24 h. The phosphorylation of ephrin B2 increased at 9 h（P〈0.05）. In addition, 2.5% XFZY-CS played a similar role as the reverse signaling activator Eph B4/Fc ranging from 0.5 to 5 μg/m L（P〉0.05）. XFZY-CS also reduced the inhibitive effect of PP2 in limited periods. Conclusion： Eph B4/ephrin B2 was the upstream signal in the process of angiogenesis and its reverse signaling was responsible for XFZY＇s effect on promoting angiogenesis.
