The Screening Study And Controlled Trials On The Anti-angiogenic Effects Based On Transgenic Zebrafish Model Traditional Chinese Medicine To Heat-clearing And Detoxifying

BackgroundIn recent years, the zebrafish as a biological research model has become a research hotspot. Zebrafish early development process, the embryo is transparent, can be directly observed in various tissues and organs. And form new blood vessels is simple and clear observation, can be clearly observed under a microscope internodes blood vessels and blood flow, and the experimental model is considered to have the characteristics of high flux and animal experiment, to study new blood vessels have congenital advantage.esistance to the new angiogenesis is a key link in the process of comprehensive treatment of liver cancer, and traditional Chinese medicine (TCM) anti-tumor angiogenesis therapy has been part of the documents confirmed. According to the literature and clinical experience, heat-clearing and detoxifying drugs of traditional Chinese medicine (TCM) is a basic method for the treatment of liver cancer. On the basis of literature review and expert consensus, we choose 6 kinds of qingrejiedu Chinese medicine for newborn screening study of angiogenesis, resistance to flila-EGFP TG strains of transgenic zebrafish embryos for biological model, using the green fluorescent protein development characteristics, water decoction of Chinese traditional medicine ingredients under the microscope of zebrafish in vivo angiogenesis inhibiting effect. For early screening by ensuring that the drug can inhibit angiogenesis effect for further quantitative test.MethodologyChoose TG strain flila-EGFP transgenic zebrafish as animal model. Will be configured fresh water extract of Chinese traditional medicine (working concentration: 200 mu g/ml) (respectively:herba violae, spreading hedyotis herb, lobelia, cordate houttuynia, the reed, wild chrysanthemum flower) and positive control drug-PTK787 (5 mu g/ml) treatment after fertilization (22 HPF) 22 hours until 48 HPF zebrafish embryos. Drug treatment after the experiment, the group will be treated as the drug concentration after embryos in asana fluorescence microscope (Nikon SMZ-1500 postures fluorescence microscope) vascular phenotype observed and photographed under, that when you take a photo for 48 HPF, among all the samples of zebrafish somite to analyze blood vessels (ISVs) generated by the impact. In the vascular screening experiments, and at the same time of the zebrafish developmental toxicity induced by various samples and death also to observe and count. After the initial screening test for drugs with anti-angiogenesis effect is ISVs quantitative count comparison and taste traditional Chinese medicine (TCM) aqueous extract of decoction liquid intervention effect of contrast.ResultsPositive control group 5 mu g/ml PTK787 can completely inhibit angiogenesis, blank control group 0.1% DMSO not inhibit angiogenesis, wild chrysanthemum flower, reed extract water leakage can be evident in the inhibition of zebrafish internode blood vessels, and herba violae, spreading hedyotis herb, lobelia, its aqueous extract has the effect of inhibition of zebrafish internode blood vessels. Six kinds of Chinese medicine extract for no cause phenomenon of zebrafish death or birth defects. Quantitative count control study, ISVs count is similar and angiogenesis inhibiting rate, suggests that wild chrysanthemum water extract and wild chrysanthemum reed leakage water extract inhibit angiogenesis effect and PTK787 is quite new, and is superior to the reed leakage water extract.ConclusionZebrafish as a new model of the screening of new blood vessels, has small volume, the advantages of the embryo, transparent, fast breeding, can be used as the research focus in the future. In the treatment of tumors need era of pursuing new and change, and more emphasis on angiogenesis in tumor growth, metastasis and the role of infiltration, choose-with the traditional Chinese medicine (TCM) of new blood vessels in order to achieve the purpose of antitumor became a hot research topic. In this study, wild chrysanthemum flower, water leakage reed extract shows more excellent resistance to the effect of new blood vessels, but also need to conduct further research, on the new angiogenesis effect of drug effect of genomics, proteomics, etc for further exploration.