A Zebrafish Screening Experiment For The Anti-angiogenesis Effect Of Ruanjiansan Chinese Medicine

Purpose:Now the medical profession believes that angiogenesis of tumor tissue could be inhibited can achieve the aim of inhibiting the growth and metastasis of tumor. Consequently, research and development of drugs in relation to cure and precaution of tumor is facing challenge in the current medical profession.We had known that angiogenesis of tumor could be inhibited by Traditional Chinese medicine. In the theory of cure tumor in Traditional Chinese medicine, also pay attention to promoting qi circulation and removing indigestion. And resolving hard lump belongs to this. Thus, it is worthy to study the Traditional Chinese medicine about resolving hard lump inhibit angiogenesis. In the experimental group, select six soft firm fights drugs to study its effect in anti-angiogenesis:Green husk, turtle shell, oysters, kelp, zhejiang fritillary and selfheal. Existing related studies show that the six drugs have antitumor effect, but lack of the study about angiogenesis on the model of zebrafish. We choose embryo of the Transgenic TG zebrafish (flila-EGFP) as biological model, Using the green fluorescent protein imaging characteristics to observe the effect of the anti-angiogenesis by Traditional Chinese medicine through microscope. And we can prove that zebrafish is feasible and applicative in Selected Chinese medicine about anti-angiogenesisMethodology:Choose the TG transgenic flila-EGFP zebrafish as a biological research model, Six typical resolving hard lump TCM (vinegar turtle, pericarpium citri reticulatae viride, oyster, kelp, selfheal, Zhejiang-Fritillaria) were decocted, concentrated, sieved and desiccated to attain the water extract. This study utilized the vascular organism research model for Flila-EGFP zebrafish, which were raised and maintained under standard conditions.22 h post-fertilization (hpf) embryos were distributed into 12-well plates for a treatment period of 26h. The TCM water extracts which were diluted in 0.1% dimethyl sulfoxide (DMSO), were added to each well at a concentration of 200 ?g/ml. The positive control was 5 ?g/ml PTK787 (vatalanib) and the negative control was 0.1% DMSO. At 48hpf larvae were tricaine anesthetized and imaged.Results:All 30 larvae treated with vinegar turtle, pericarpium citri reticulatae viride and PTK787 showed angiogenesis defects. Embryos treated with vinegar turtle and pericarpium citri reticulatae viride present a lower number of complete ISVs. TCM did not show neurotoxicity or organ toxicity in the zebrafish. No death or teratogenesis was seen in the TCM.Conclusion:The present research demonstrates that vinegar turtle and pericarpium citri reticulatae viride have anti-angiogenic effects in zebrafish. No death or teratogenesis was seen in the TCM.