中 文 摘 要本研究利用台灣栽培最普遍之松杉靈芝( Ganoderma tsugae, Murrill )為材料,針對其子實體 (fruiting body) 中分段萃取出之三種 萃取液,經超微過濾 (Ultrafiltration),半透膜 (semipermeable membrane) 透析 (dialysis) 及冷凍乾燥等步驟而得到三種多醣 [分別 為1. Fraction I (water soluble polysaccharide,簡稱F-I) 2. Fraction II (alkali soluble polysaccharide,簡稱F-II)及 3. Fraction III (第三類polysaccharide,簡稱F-III)三個部分] ,將這三 種多醣進行成分分析及各種生理活性的測試。包括了: I.成分分析, 1.以 TLC,HPLC 方法得知其單糖 (monosaccharide) 組成;2.以 Gel filtration 方法求得其分子量 (molecular weight) ; 3.總糖含量測定 ; 4.總氮含量測定; II生理活性 (主要是針對其免疫及抗腫瘤活性): 1.體內抗人工轉移腫瘤試驗:靜脈注射投予三種多醣,以 PI 染色法偵測 S-180 cell 之 DNA 特性,利用 S-180 cell DNA 含量與 normal lung cell DNA 含量之不同,以流動細胞分析儀 (Flow cytometer) 分析腫瘤 轉移肺臟中 S-180 tumor cell 與投藥之間的關係; 2.觀察且計算不同 組之肺表面瘤結節 (nodule) 數; 3.測試三種多醣對末稍 (peripheral) 血液 T,T4,T8,B 及 macropharge 比率之影響。結果顯示, F-I 及 F-II 在化學組成上並無多大差異,也和 PSK 相差不多,均為 glucose 與 N- acetyl-D-glucosamine 或 Glucosamine 之共聚物;而 F-III 則以 Glucosamine 為主 (約60%)。此外在對抗人工轉移腫瘤之效果上,以 G2+ M(%) 之 S-180 細胞而言,則以 F-III (38.28%) > F-I (64.30%) > F- II (79.84%) > Control (100%)。
AbstractIn order to completely utilize the polysaccharide isolated from the fruiting body of ''Ling Zhi'', this study used the most common cultured species in Taiwan, the mushroom Songshan lingzhi (Ganoderma tsugae,Murrill), as its material. We extracted three polysaccharides by different processes such as ultrafiltration, semimembrane dialysis, and lyophilization. The purpose of the study is to compare the chemical and antitumor properties of the polysaccharide produced by 1. F-I (water soluble polysaccharide), 2. F-II (alkali soluble polysaccharide), and 3. F-III (the deacetylated cell wall skeleton). The chemical analysis included total sugar assay, sugar component, ratio of the sugar component, and molecular weight. The methods involved in the analysis are : TLC, HPLC, Gel-filtratiion, and Acid hydrolysis. Antitumor activities of these polysaccharides were assayed by challenging Sarcoma 180 tumor cells to Balb/c mice and prior to which the polysaccharides were administrated into cadal vein, intravenously. The pulmonary nodules of S-180 induced by metastasis were counted by flow cytometer and naked eyes. In addition, the lymphocyte subsets in peripheral blood were measured in respect to the ratio of T, T4, T8, B, and Macrophage. In this experiment, we tried to utilize the residue of Ling Zhi which is used to be dumped. In the whole process, two additional polysaccharides were produced in large quantity and we expect these polysaccharides functioned as well as that from water soluble one. These results indicate that F-I and F-II had no much difference in their components. They were the polymers of glucose and N-acetyl-D-glucosamine (or glucosamine). The major component of F-III is glucosamine (60%). In addition, to the G2+M(%) phase of murine S-180 sarcoma cells, the inhibitory effect is F-III (38.28%) > F-I (64.30%) > F-II (79.84%) > control (100%).