Research On Anti-angiogenic Activity Of Chonglou Based On Zebrafish Model

In recent years,the rising morbidity or mortality of tumor has become one of the key problems in medical research.Pathological surveys show that angiogenesis is a necessary condition for tumor's growth.Inhibition of angiogenesis is a good target for cancer treatment.Most anti-angiogenesis drugs used in clinic take effects through the role of single target which is associated with high costs,high-risk to produce the drug resistance and relapse of tumor cells.These problems impose restrictions on their clinical application seriously.Therefore,to find new anti-angiogenesis drugs has become an urgent problem in the field of cancer treatment.After thousands of years of clinical evidence-based,traditional Chinese medicine has many advantages,such as low price,multiple ingredients,multiple targets and so on.This is an important source for discovery of new anti-angiogenesis prodrugs.As one of traditional classes in Chinese herbal,heat-clearing and detoxicating drugs,which is commonly used in the treatment of cancer patients,has become the first choice for anti-angiogenesis drugs screening.In this study,transgenic zebrafish line Tg(flk1:mCherry)was used to screen the anti-angiogenesis effect of 31 herbal extracts during the same concentration range,then Paris polyphylla was selected as the study object subsequently.The tumor IOD value and SIV of the transgenic zebrafish xenografts were used as indicators to evaluate the anti-angiogenesis activity of the extracts.The anti-angiogenesis effect of saponins from Paris polyphylla was further studied.The results of this study will provide references for further anti-tumor mechanism research and new anti-tumor drug development of Paris polyphylla.The main contents of this study are described as followsChapter 1:Transgenic zebrafish line Tg(flk1:mCherry)was used to study the effects of 31 Chinese herbs on the intersomatic vessels angiogenesis from 6 hpf to 72 hpf in the same concentration range(50?g/mL?100?g/mL?200?g/mL),while 100 ng/mL PTK787 was used as positive control.The results showed that Potentilla Ampelopsis japonica and Potentilla chinensisSeringe promote the formation of intersomatic vessels of zebrafish at different concentrations.Andrographis paniculata,Isatisindidigotica,Violayedoensis,Crematrae Pseudobulbus Pleiones Pseudobulbus,SophoraeTonkinensis Radix Rhizoma inhibited the intersomatic vessels angiogenesis of zebrafish in different concentrations with growth inhibition or developmental malformation of zebrafish larva at the same time.Paris polyphylla exhibited good inhibiting effect on the intersomatic vessels of zebrafish,while rarely affect the growth or development of zebrafish.In chapter 2,the anti-angiogenesis effects of different extracts of Paris polyphylla were studied on transgenic zebrafish model,with the IOD value of tumor and the area of inferior intestinal vessel as the index.Results showed that at the concentration of 100ng/mL,200ng/mL,400ng/mL,the drug could inhibit both subintestinal blood vessel angiogenesis and xenografted human hepatoma tumor growth in transgenic zebrafish in a concentration-dependent manner.Chapter 3:polyphyllin ?,?,?,?,? and H from Paris polyphylla were compared at physiological anti-angiogenesis activity on zebrafish model.The results showed that Paris polyphylla polyphyllin H promoted the angiogenesis of zebrafish only at concentration of 100ng/mL.Polyphyllin ? inhibited the intersegmental vessels of zebrafish at 50ng/mL and 100 ng/mL,while polyphyllin ? inhibited the intersegmental vessels of zebrafish at concentration of 200 ng/mL.At the concentrations of 100ng/mL,200ng/mL and 400 ng/mL,polyphyllin ? and ? could inhibit the growth of tumor and the development of intersegmental vessels of zebrafish.This study showed that some of the heat-clearing and detoxicating drugs of traditional Chinese medicine had effects on angiogenesis,especially the anti-angiogenesis activity.The combination of transgenic zebrafish model and xenograft model can observe the changes of blood vessels and tumor blood vessels in zebrafish at the same time.This study provides in-vivo evidence for tumor treatment of Paris polyphylla and lays a foundation for the development and application of Paris polyphylla ? and ?.