Chlorogenic Acid-enriched Extract From Ilex Kudingcha C.J.Tseng Inhibits Angiogenesis In Zebrafish And Its Mechanism

Kudingcha is a particularly bitter-tasting tea that has been widely used in China to cure disperse wind heat and itching eyes,and to clear blood toxins.Kudingcha is considered of value for its potential anticancer effects,which are attributed to the presence of characteristic ingredients.We previously demonstrated that chlorogenic acid(CGA)derivatives from kudingcha extract(KDCE)exhibit dose-dependent anti-angiogenesis activity in intersegmental vessels(ISV)and induce apoptosis in vascular endothelial cells in zebrafish,but the mechanisms remain unknown.Herein,ten differentially abundant proteins associated with extracellular signal-regulated protein kinases(ERKs)were identified following exposure to CGA in zebrafish embryos at 52 hour post-fertilisation(hpf)using isobaric tags for relative and absolute quantitation(i-TRAQ)analysis.Among the ten proteins,CGA was successfully docked into the active site of MAP4K5 alone by Molecular Operating Environment(MOE)analysis.KDCE and CGA also induced ERK phosphorylation in zebrafish.These results indicate that KDCE and CGA induce anti-angiogenesis via ERK phosphorylation,and CGA interacts with MAP4K5 in the response to apoptosis in vascular endothelial cells.Importantly,our findings suggest that KDCE is a potential anti-angiogenic reagent and food additive.