中文摘要

血紅密孔菌( Pycnoporus sanguineus )栽植體之成份分析與抗發炎活性研究

Studies on chemical constituents and anti-inflammatory activity of cultivated Pycnoporus sanguineus

中文摘要

血紅密孔菌 (Pycnoporus sanguineus (Fr.) Murr) 泛佈於熱帶和亞熱帶地區，為一種木材腐 朽菌，野生子實體生長速度較為緩慢，因此造成研究材料之不足，本研究針對培養之菌絲體進行 生成之生物量及化學成分研究。結果顯示血紅密孔菌在培養 49 天後，菌絲乾重達最高產率 16.13±2.10 g/l，而多醣體產量在培養 21 天時，達 1.02±0.40 g/l 最高。培養第 28 天時 乙醇萃取物產量為 6.10±1.90 g/l 最高。而所含多醣體之單醣組成分析，結果顯示以岩藻醣 (fucose)、葡萄糖 (glucose)及甘露糖 (mannose) 為主要單糖組成，其含量分別為

64.81±0.15,18.70±0.05, 17.56±0.03 μmol/g PS。針對血紅密孔菌菌絲體進行分離純 化，共得到四個麥角甾醇，分別為：ergosta-7,22-dien-3β-ol (ST-1: 1),3β,5α-dihydroxyergosta-7,22-dien-6-one (ST-2: 2), 3β,5α,9α-trihydroxy-ergosta-7,22-dien-6-one (ST-3: 3)及 3β,5α,6α-trihydroxyergosta-7,22-diene (ST-4: 4)。ST-2, ST-3, ST-4 三個化合物是首 次自血紅密孔菌中分離得到之已知化合物。藥理活性測試以 LPS 誘導老鼠微神經膠細胞株 (BV-2) 產生 NO 的抗發炎模式篩檢中，乙醇粗萃物在 50 μg/ml 濃度下，抑制率達 87.5 %， 具有極顯著抑制 NO 產生的效果。乙醇粗萃物之劃分，乙酸乙酯層在 20 μg/ml 濃度下，抑制 率達 87.5 %，正丁醇層 20 μg/ml 濃度時，抑制率 45.5 %。經單離後之純化合物 ST-1 對 LPS 誘導之微神經膠細胞株在 15 μM 濃度具有 43.5 % 的抑制率。

英文摘要

Pycnoporus sanguineus (Fr.) Murr is a slow-growing saprophytic fungus which causes decay of certain types of wood in the forests of tropical and subtropical areas. Because of the material is limited on the island and poor growth rate in nature, liguid culture of the fungus was

attempted. The results showed that the fungus was cultured for 49 days to reach the maximal dry weight of 16.13±2.10 g/l. The yield of polysaccharides (PS) of P. sanguineus for 21 days was 1.02±0.40 g/l. The yield of ethanolic extract of mycelial of P. sanguineus for 28 days was

6.10±1.90 g/l. Compositional analysis PS of P. sanguineus showed that fucose, glucose and mannose were the predominant sugars in the value of 64.81±0.15, 18.70±0.05 and 17.56±0.03 μmol/g PS, respectively.

Bioassay-guided fractionation was led to the isolation of four ergostane skeleton type steroids, including ergosta-7,22-dien-3β-ol (ST-1: 1),

3β,5α-dihydroxyergosta-7,22-dien-6-one (ST-2: 2),

3β,5α,6α-trihydroxyergosta-7,22

-diene (ST-4: 4) 。All were known compounds. From the isolates,

ergosta-7,22-dien-3β-ol was isolated from P. sanguineus previously. The others were the first isolation from mycelial of P. sanguineus. Their structures were determined by 1H, 13C, 2D NMR, and MS spectral

analyses. The bioassay-guided fractionation of the crude ethanolic extract showed significant inhibition of LPS induced NO production in BV-2 cells up to a

concentraction of 50 μg/ml for 87.5 %. EA-layer extract was also showed significant inhibition of LPS induced NO production in

BV-2 cells up to a concentraction of 20 μg/ml for 87.5 %. BuOH-layer extract of 20 μg/ml for inhibition of 45.5 %. Compound ST-1 of the inhibition at the