Effect of EphB4/EphrinB2 reverse signal on angiogenesis induced by Xuefu Zhuyu Capsule () containing serum in human microvascular endothelial cell 1.
OBJECTIVE
To evaluate the effect of Xuefu Zhuyu Capsule ()-containing serum (XFZY-CS) on EphB4/ephrinB2 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 EphB4/ephrinB2 and the reverse signal were determined by Western blot and real-time quantitative polymerase chain reaction, respectively; we also confifirmed the results through activating and inhibiting the reverse signal by EphB4/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 EphB4-mRNA at 12 h (P<0.05), and down-regulates its expression at 24 h (P<0.01). Protein expression of EphB4 was apparently up-regulated at 12 h and down-regulated at 24 h. The phosphorylation of ephrinB2 increased at 9 h (P<0.05). In addition, 2.5% XFZY-CS played a similar role as the reverse signaling activator EphB4/Fc ranging from 0.5 to 5 μg/mL (P>0.05). XFZY-CS also reduced the inhibitive effect of PP2 in limited periods.


CONCLUSIONS
EphB4/ephrinB2 was the upstream signal in the process of angiogenesis and its reverse signaling was responsible for XFZY's effect on promoting angiogenesis.
Zhongshan Hospital, Fudan University, Shanghai, 200032, China.
Jing-Si
JS
Zhang
College of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, 350108, China.
Yi-Zheng
YZ
Wang
College of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, 350108, China.
Ya-Qiong
YQ
Hu
College of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, 350108, China.
Fan
F
Lin
College of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, 350108, China. 976558160@qq.com.
Dong
D
Gao
Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Jun
J
Song
Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA02215, USA.
Ted J
TJ
Kaptchuk
Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, 100091, China.
Ke-Ji
KJ
Chen
D000328:Adult; D002214:Capsules; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D036388:Ephrin-B2 / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D055806:Microvessels / Q000473:pathology*; D008875:Middle Aged; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000235:genetics; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D036224:Receptor, EphB4 / Q000235:genetics / Q000378:metabolism*; D044967:Serum / Q000378:metabolism*; D013997:Time Factors; D055815:Young Adult
D002214:Capsules; D004365:Drugs, Chinese Herbal; D036388:Ephrin-B2; D012333:RNA, Messenger; C513826:Xue-Fu-Zhu-Yu decoction; D036224:Receptor, EphB4; D010727:Phosphoric Diester Hydrolases
Xuefu Zhuyu Capsule; angiogenesis; ephrinB2/EphB4 reverse signal