- 學位論文
探討甘草萃取物影響血管生成之能力
Assessment of pro-angiogenic ability of Liquorice extract
摘要
過敏煎主要是由甘草、五味子、銀柴胡、防風、烏梅五種中藥材所組成，此藥物具有抗過敏及抗發炎的作用，其中甘草根據先前研究發現，甘草水萃物具有有效抗血管新生及抗腫瘤的活性，推測甘草與血管生成有密切的關係，而本篇論文主要研究目的為探討甘草萃取物是否具有影響血管增生之能力。透過斑馬魚Alkaline Phosphatase staining實驗，發現浸泡過敏煎及甘草後之斑馬魚胚胎的SIV branch points在(basket inside: 對照組 vs. 過敏煎, 1.00±0.18、2.00±0.30； basket outside: 對照組 vs. 過敏煎, 0.17±0.06、0.67±0.12)；(basket inside: 對照組 vs. 50 ppm甘草, 1.28±0.13、1.87±0.22； basket outside: 對照組 vs. 50 ppm甘草, 0.81±0.09、1.42±0.13)，皆有明顯促進血管增生的現象。接下來使用基因轉殖斑馬魚Tg(fli1:EGFP) 觀察血管增生的效果，發現在浸泡過敏煎後caudal vein plexus血管面積增加3.6%，也發現有促進血管增生的現象。最後在基因層次的研究發現， flt1、nrp1a (皆為血管內皮生長因子受體)、cdh5(細胞連接標記)基因表現量分別為對照組的 1.99倍、0.86倍及1.21倍。根據上述實驗結果，可得知過敏煎與甘草萃取物確實具有促進血管增生的能力。
並列摘要
Guo Min Jian (GMJ) is mainly composed of five kinds of Chinese medicine include Liquorice, Wu wei zi, Yin chai hu, Fang feng, and Wu mei. This Chinese medicine has anti-allergic and anti-inflammatory effects. According to previous paper, the water extract of Liquorice has anti-angiogenic and anti-tumor activity, it is inferred that Liquorice is closely related to angiogenesis. The main purpose of this thesis is to explore the pro-angiogenic ability of Liquorice extract. Results of Alkaline Phosphatase staining experiments, showed that the SIV of GMJ and Liquorice-treated zebrafish embryos had more branch points (basket inside: mock: 1.00±0.18 vs. GMJ-treated: 2.00±0.30；basket outside: mock: 0.17±0.06 vs. GMJ-treated: 0.67±0.12)；(basket inside: mock: 1.28±0.13 vs. 50 ppm Liquorice-treated: 1.87±0.22；basket outside: mock: 0.81±0.09 vs. 50 ppm Liquorice-treated: 1.42±0.13). Results of fluorescence recording of transgenic zebrafish Tg(fli1:EGFP) showed that GMJ-treated embryos displayed more broadly fluorescence expressions in the caudal vein plexus (the area of blood vessel increased by 3.6%) compared to no treatment mock group, indicating that both GMJ and Liquorice possess pro-angiogenic effects. Finally, from the genetic points of view, it was found that the expressions of flt1(VEGFR-1) and cdh5 (cell adhesion marker) genes in the GMJ-treated groups increased to 1.99 and 1.21 folds, but the expression of nrp1a (VEGFR) decreased to 0.86 folds. On the basis of these observations, we proposed that GMJ and Liquorice extract have pro-angiogenic effects.