Angiogenesis was the formation of new capillary blood vessels from pre-existing vessel, which was an important biological process in physiological and pathological conditions. A variety of diseases such as cancer, diabetic retinopathy, rheumatoid arthritis, ischemic heart disease, chronic wound were associated with dysregulation of angiogenesis. ADTM was new danshensu derivative, synthesized by chemical modification of Danshensu (DSS) and tetramethylpyrazine (TMP), which were active ingredients of Traditional Chinese medicine Salvia miltiorrhiza and Ligusticum wallichii, respectively. These two herbs were widely used for the treatment of ischemic heart and cerebral vascular disease. In present study, we found that ADTM significantly exhibited pro-angiogenic activity in zebrafish, whereas the underlying mechanism of angiogenesis effect had not been addressed. We investigated angiogenic effect of ADTM using in vivo and in vitro. The in vivo angiogenic effect of ADTM was studied using transgenic Tg(fli-1a:EGFP)y1 zebrafish embryos by observing the presence of ectopic vessels in the subintestinal vessels (SIV) and recuse iii effect of chemical-induced blood vessel loss in intersegmental vessels (ISV) and dorsal longitudinal anastomotic vessel (DLAV) under fluorescence microscopy. ADTM did not exhibit angiogenesis activity in endothelial cell proliferation and capillary tube formation assays, but it significantly reversed VRI-induced blood vessel loss in zebrafish. Furthermore, ADTM also could reverse VRI-induced down-regulation of mRNA expression of VEGFR1 and VEGFR2. This present study proposed the advantage of zebrafish over endothelial cell for screening angiogenic compounds because some compounds were only active in in vivo but inactive in in vitro. Taken together, these results suggested that ADTM was novel angiogenic agent and had potential to develop as a therapeutic agent for diseases associated with imbalance angiogenesis such as ischemia heart disease.

血管新生在很多生理過程中扮演著重要的角色，並且與很多疾病有關，例如癌症、 糖尿病視網膜病變、類風濕性關節炎、缺血性心臟病、慢性傷口。ADTM，一種 新的丹參素衍生物，主要由丹參素和川芎嗪為基本化學結構，經過化學修飾後合 成的衍生物。丹參素和川芎嗪分別為傳統中藥丹參和川芎的主要活性成份之一， 這兩種中藥在中國數千年以來用作治療缺血性心腦血管疾病。在本次研究中， 使用轉基因 Tg(fli-1a:EGFP)y1 斑馬魚研究 ADTM 促血管新生作用和其分子機 制。 實驗結果發現，ADTM 在斑馬魚模型上具有很強的促血管新生作用。ADTM 能 能夠逆轉血管內皮細胞生長因子受體抑制劑 (VEGF receptor tyrosine kinase inhibitor, VRI) 誘導體節間血管 (ISV) 和 背部縱向血管 (DLAV) 缺失，且呈劑 量依赖關係。同時通過基因表達實驗結果發現，ADTM 能夠逆轉 VRI 引起的 VEGFR1 和 VEGFR2 表達下調。然而，在血管內皮細胞體外模型上没有顯著的 促血管新生作用。本研究提示，ADTM 在斑馬魚體內模型上具有很強的促血管 新生作用。另一方面，該研究表明，使用斑馬魚模型篩選血管新生的化合物優越 於血管內外細胞模型，且會减少假陰性結果。 關鍵字: 血管新生，斑馬魚，ADTM，丹参素，川芎嗪，血管內皮細胞，血管內 皮細胞生長因子