- 學位論文
當歸與黃耆多醣對人類臍帶血造血幹細胞之 增生與分化之影響
Effects of Angelica sinensis and Astragalus membranaceus polysaccharides on proliferation and differentiation of hematopoietic stem cells from umbilical cord blood
摘要
本論文乃研究當歸與黃耆對臍帶血中造血幹細胞 (CD34+ cells) 之增生與分化之影響，分三部份進行。第一部分的實驗中，我們確認多醣為當歸水萃液中與造血功能相關的活性成分，並以陰離子交換層析分離當歸粗多醣，得到四個不同的區分，其中 F 2 區分為主要的活性分布，能促進 GM-CSF、 IL-3 分泌，幫助 CFU-GM 的形成，其分子量介於 51.28 kDa 與 8.40 kDa 間，主要由阿拉伯糖、半乳糖及半乳糖醛酸所組成。第二部份我們將當歸與黃耆配伍使用，來探討藥材經配伍使用後，能否增進其藥效。結果顯示，多醣亦為黃耆成分中與造血活性相關的有效成分。雖然黃耆在配伍的比例中為主體，但其活性遠不如當歸多醣，但黃耆多醣能提供或是造成協同作用，導致兩藥材在配伍使用後，能同時促進 GM-CSF、 IL-3 及 M-CSF 等 HGF 的分泌，助於 CFU-GM 及 BFU-E 兩種集落的形成，並顯著提高 BFU-E 所佔的比例。第三部份，乃探討 F 2 區分及配伍藥方，在抑制劑 Adriblastina 的存在下，對於造血幹細胞提供之保護作用，結果顯示，配伍藥方比 F 2 區分能更有效的減緩抑制劑造成之傷害。
並列摘要
The aim of this research was to investigate the effects of Angelica sinensis (AS) and Astragalus membranaceus (AM) polysaccharides on proliferation and differentiation of hematopoietic stem cells (CD34+ cells) from umbilical cord blood. This research was divided into three parts. In the first part, we confirmed that polysaccharides was the major hematopoietic component in the AS water extract. The Angelica sinensis polysaccharides (ASPS) were fractionated into four fractions by anion-exchange chromatography, and F 2 fraction had the highest hematopoietic activity among four fractions. The F2 fraction could stimulate secretion of hematopoietic growth factor (HGF), such as GM-CSF and IL-3, and promote colony-forming unit of granulocyte-macrophage (CFU-GM) formation. The molecular weight of F 2 fraction was between 51.28 kDa and 8.40 kDa, and composed of arabinose, galactose and galacturonic acid. In part II, we used the decoction - Danggui Buxue Tang (DBT), which is composed of AS and AM with weight ratio 1：5. We compared the hematopoietic activity of DBT, AS and AM, and found that the combination of AS and AM had synergizing on hematopoiesis, and AS and AM polysaccharides were the active component in DBT. DBT induced all HGF that we detected, including GM-CSF, IL-3 and M-CSF. Compared to F2 fraction, DBT could stimulate more kinds of HGF production, and this is the probably reason why DBT had higher hematopoietic effect than F2. DBT not only promoted the formation of CFU-GM but also promoted Burst-forming unit erythrocyte (BFU-E) colony formation. In part III, we compared the protective effect of F2 fraction and DBT against the damage caused by the chemotherapy drug Adriblastina. It was found that both the F 2 fraction and DBT rescued CD34+ cells viability and protected the colony formation ability of CD34+ cells. In this damage model, DBT was found to have better effects than F 2 fraction.