Turtle Shell,Green Husk Which Inhibit The Zebrafish Anaioaenesis Regulate The VEGF,Notch/D114 Pathways

Background and AimsTherapies targeted to blood vessels,angiogenesis,shown promising including antiangiogenic therapy otentials in the treatment of mostdiseases such as cancer, cardiovascular disease.our understanding of the molecular mechanisms for vascular development and angiogenesis remains incomplete. Arteries and veins are structurally and functionallydistinct organs critical to embryonic development and adult life. The development of these vascular systems is one of the earliest steps in embryogenesis.Vascular endothelial growth factor VEGF family is considered to be the most important secreted angiogenic factors,in the embryonic development of cardiovascular system and the process of tumor angiogenesis play an important role.In the process of vascular development closely related to VEGF, Notch signal pathway, although can play a different role independently,but there are interactions between them.Traditional Chinese medicine treatment of cancer has a history of thousands of years, has accumulated a wealth of clinical information Ruanjian sanjie is a major solution treatment of tumors,for this class pf drugs in the aspect of antiangiogenic effect is worthing further study. Flila-this experiment by TG strains of EGFP transgenic zebrafish as a model, using the green fluorescent protein imaging characteristics, water extract from Chinese traditional medicine ingredients under the microscope of zebrafish in vivo angiogenesis inhibiting effect, and to further study has the ability of anti angiogenesis effect of two Chinese medicine regulation of VEGF, Notch/Dll4 pathways.Methods:1?Using TG lines flila-EGFP transgenic zebrafish as a biological model. The fresh configured herbal extracts (respectively:turtle shell,green husk) leakage treatment after fertilization of zebrafish embryos.After the drug treatment was finished, the embryos of each group were observed and photographed under the fluorescence microscope,to analyze the effects of various samples on the formation of the vascular (ISV) in the body of zebrafish.2, Use of real-time fluorescence quantitative PCR technology to detect zebrafish embryos by the turtle shell,green husk treatment, VEGF signal pathway in VEGFaa/VEGFR1/VEGFR2 and the expression of D114/Notchla/HEY2/EFNB2a in the Notch signaling pathway, study of these two signaling pathways in the physiological role in the process of embryonic development.Results:1, The extracts of turtle shell,green husk intervention with experimental group zebrafish embryos,the fluorescence expression of ISVs and DLAV were different, suggesting that the effect of inhibiting angiogenesis of zebrafish.2, The extract of turtle shell,green husk can significantly increase the expression level of D114 in zebrafish embryos(P<0.05).The extract of turtle shell,green husk can significantly increase the expression level of Notch1? in zebrafish embryos, the role of turtle shell,green husk is particularly prominent(P<0.01). The extract of turtle shell,green husk can significantly increase the expression level of HEY2 in zebrafish embryos, the role of green husk is particularly prominent(P<0.01). The extract of green husk can significantly increase the expression level of EFNB2a in zebrafish embryos, (P<0.01).speculated that the two kinds of extracts inhibited of angiogenesis in zebrafish embryos is related to the up regulation of DLL4/Notchla/HEY2 gene expression.3, The extract of turtle shell,green husk can significantly increase the expression level of VEGFaa in zebrafish embryos, the role of green husk is particularly prominent(P<0.01). The extract of turtle shell,green husk can significantly increase the expression level of VEGFR1 in zebrafish embryos, the role of turtle shell,green husk is particularly prominent(P<0.01). The extract of turtle shell,green husk can significantly increase the expression level of VEGFR2 in zebrafish embryos, the role of green husk is particularly prominent(P<0.01). So,we presumably two kinds aqueous extract can activate the VEGF pathway.4, The level of expression of VEGFaa gene expression level increased significantly compared with blank control group(p<0.05);The level of expression of VEGFR1/VEGFR2 mRNA could be inhibited by vatalanib(P<0.01).Conclutions:1,The extract of turtle shell,green husk can have anti-angiogenic effects in zebrafish.2, The extract of two kinds of Chinese medicine have effects in inhibition of angiogenesis related to up regulation of D114/Notchla/HEY2/EFNB2a gene expression in zebrafish embryos;3, The water extract of turtle shell, green husk which inhibit angiogenesis can activate the VEGF signaling pathway, there may be feedback regulation.between VEGF and Notch-D114 signaling pathway.