Primary screening of Chinese materia media for angiogenesis modulating activities using zebrafish embryos
Student thesis: Master's Thesis
Zebrafish-based screening combines the advantage of higher through-out analysis and high relevance to humans. Chinese material medica (CMM) are likely to possess natural products of potential therapeutic values as they have demonstrated their effectiveness in the treatment of ailments and diseases over their long history of empirical use. They are becoming increasingly important in modern health care because they can reduce medical costs and lead to clinical success in a different range of conditions. Angiogenesis is a natural process of the growth of new blood vessels. It is believed that both too much and too little angiogenesis are associated with cancer, coronary artery disease and numerous other diseases. The research was conducted by performing screening experiments using whole-animal models and data mining. The screening was a phenotype-based analysis. It was performed by scoring angiogenic activity of zebrafish embryos. This was achieved by using in vivo assay and screening tool. The assay was carried out by the addition of CMM to zebrafish embryo medium. Alkaline phosphatase activity was used as a screening tool to visualize vascular pattern in zebrafish embryos. A database was developed by storing information from traditional knowledge and experimental data in computerized format. The data of screened CMM included components of formulae and the botanical names of herbs, etc. The formation or malformation of the vessels of each fish during development was recorded. New variables derived from existing recorded parameters, e.g. survival rate. Then, additional data exploration was carried out. Finally, three visualizations and one application were developed to show relationships after data mining. There were totally 178 CMMs screened. The screening system successfully identified 4 formulae, 11 single herbs and 11 single compounds inducing anti-angiogenesis and 3 formulae inducing pro-angiogenesis. It was reproducible as the enzymatic staining assay was quick and easy. The experiments was continued to screen more CMMs and the secondary screening was processed by my teammates. In addiction, it was successfully completed in a short period and provided a high hit rate. The data mining provided a platform for studying and understanding CMM. It was expandable and was searchable by various parameters. It functioned via the World Wide Web that allowed multiple users to access worldwide. The research bridged the gap between CMM and angiogenesis.
- Animal models, Zebra danio, Embryos, Physiology, China, Materia medica, Neovascularization