當歸粗多醣對髓系造血功能的影響

Abstract

本研究以當歸水萃物為原料，區分為小分子 (< 300 Da)及大分子 (> 300 Da，酒精沉澱粗多醣)，以探討當歸是否具有傳統中國醫藥所稱之「補血」功效。本研究以 in vitro及 in vivo兩種方式進行。前者利用取自小鼠之造血幹細胞進行實驗，觀察樣品是否有刺激血球細胞集落增生之效果；後者則以急性失血性貧血的動物模式 (BALB/c小鼠)探討當歸區分物對造血功能之影響。 研究結果發現，當歸水萃物中小分子成分不具有刺激血球細胞集落數增加的效果。但是，以大分子之多醣先行刺激脾臟細胞，產生條件培養液，再以含細胞激素之條件培養液刺激造血幹細胞，則能顯著的增加血球細胞集落數 (p < 0.05)。所形成的集落的集落型態主要有顆粒球集落 (granulocytic colony)、顆粒球-單核 (granulocyte and monocyte colony)及顆粒球-巨噬細胞混合型集落 (granulocyte and macrophagic colony)。最後，在以動物模式評估造血功能方面，發現當歸粗多醣並不會影響小鼠正常生長情形以及主要器官之重量；低劑量時，當歸粗多醣顯著提升失血後小鼠之血紅素值 (p < 0.05)；低、高劑量皆具有刺激脾臟細胞增生的能力 (p < 0.05)，並且於高劑量組中可顯著增加細胞激素 GM-CSF及 IL-6 (p < 0.05)；最後，低、高劑量皆可顯著的增加集落數目 (p < 0.05)。

Angelica sinensis (Oliv.) Diels is a well-known herb belonging to family Umbelliferae, and is traditionally used for the treatment of anemia. The aim of this study was to investigate hematopoietic effect of extracts of Angelica sinensis (AS). For in vitro studies, the hematopoietic stem cells, collected from BALB/c mice were treated with AS, to observe the colonies-forming activity. For in vivo studies, acutely bleed BALB/c mice were administered polysaccharides which were precipitated with EtOH (ASPS) orally, to assess the hematopoietic activity of Angelica sinensis extracts. The AS extract was subjected to membrane filtration using hollow fiber membrane module with molecular weight cut-off of 300 Da. Spleen cells from mice was treated with the permeate containing components of AS with molecular weight less than 300 Da (ASsp) and the AS polysaccharides which were precipitated with EtOH from the retentate (ASPS). Then, the spleen condition medium (ASsp-SCM, ASPS-SCM) was collected. It was found that the ASsp-SCM could not promote colony-forming in bone marrow cells. However, bone marrow cells treated with ASPS-SCM showed significant (p < 0.05) increase in colony formation as compared to AS-SCM and ASsp-SCM. Morphologic analysis of the colonies formed by ASPS-SCM showed three cell types, including granulocytes, granulocytes /monocytes and granulocytes /macrophages. The result of in vivo study showed that, the low dose of ASPS significantly (p < 0.05) increased the production of hemoglobin, while the high dose of ASPS significantly (p < 0.05) increased the levels of GM-SCF and IL-6 as compare with control and low dosage. Also it was observed that the high dose of ASPS significantly (p < 0.05) proliferated spleen cells. This herbal medicine activates immature blood cells, possibly by suppressing cytokine secretion, thereby promoting hematopoiesis. Thus the in vitro and in vivo studies have proved that Angelica sinensis has significant hematopoietic and proliferative activity and can be used in anemia treatment.