野生食用菇類作為藥品與化妝品潛在應用性之研究

Abstract

在第一章中，由國立科學博物館植物研究部保坂健太郎博士提供的 34 種野生食用菌萃取物，經由 TLC、抗菌活性和抗氧化活性試驗進行蘑菇篩選，本實驗選擇了生物活性相對較高的 Albatrellus confluens 進行研究。將其乙酸乙酯萃取物利用 HPLC、MS 及 NMR 等方法分離並純化出 Meroterpenoids Grifolin 及 Neogrifolin，而這兩種化合物的結構與 Podogigant A 等化合物相似，Podogigant A 曾在文獻中被提及可提升真菌劑 amphotericin B (AmB) 的抗真菌活性，因此後續實驗與北里大學合作研究 meroterpenoids Grifolin 及 Neogrifolin 兩種化合物的活性。結果顯示 2 μg/mL 和 4 μg /mL 的組合對帶有念珠菌病病原的白色念珠菌具有 4 倍AmB 增效活性，而 8 μg /mL 的組合則具有 8 倍的 AmB 增效活性。因此將 meroterpenoids Grifolin 及 Neogrifolin 與抗生素結合，可減少所需劑量，並有助於將副作用減至最低。 在第二章中，自 30 種野生食用菌萃取物，包含先前研究曾使用的蘑菇、文獻未曾提及與分離成天然產品的蘑菇、長期被當作藥用蘑菇的蘑菇 (例如靈芝 Ganoderma lucidum)，以及食用蘑菇 (例如日本白松露 Tuber japonicum) 等，這些蘑菇萃取物在台灣大學進行美白活性測試。首先測試酪氨酸酶的抑制活性，酪氨酸酶有助於產生黑色素，而黑色素是使皮膚變黑的成分。結果發現，其中七種蘑菇具有相對較高的抑制活性。接著使用小鼠身上產生黑色素的 B16-F10 黑色素瘤細胞進行活性抑制與毒性測試，結果發現 Ganoderma lucidum、Boletopsis grisea、Pleurotus ostreatus、Grifola frondosa、Albatrellus confluens 和 Calvatia craniiformis 等六種蘑菇在無毒濃度範圍內具有美白活性。最後使用斑馬魚進行測試，結果顯示，這六種蘑菇的萃取物皆能抑制色素沉澱，且不會對生物造成不良影響。因此這些蘑菇具有開發新的、安全的、天然的美白化妝品材料的潛力。

In the 1st chapter, several species of mushrooms were selected by screening 34 wild edible mushroom extracts provided by Dr. Kentaro Hosaka of the National Museum on Nature and Science, Plant Research Department, Japan, using TLC, antibacterial activity, and antioxidant activity tests, according to previous research conducted by the same laboratory. In the initial experiments, Albatrellus confluens, which showed relatively high biological activity among them, was selected for study. The ethyl acetate extract was divided into several fractions, and the meroterpenoids grifolin and neogrifolin were isolated, purified, and elucidated using HPLC, MS, and NMR. Since the structures of these two compounds were similar to those of compounds such as podogigant A, which has been reported to enhance the antifungal activity of the fungal agent amphotericin B (AmB), I investigated their activity in collaboration with Kitasato University. The results showed that 2 and 4 μg/mL combination showed 4-fold and 8 μg/mL combination showed 8-fold AmB potentiating activity against Candida albicans, the cause of candidiasis. Combining those two compounds with antibiotics may reduce the required dosage and minimize side effects. In the 2nd chapter, extracts were obtained from 30 wild edible mushrooms, including several mushrooms used in previous studies, mushrooms that have not been reported and isolated as natural products, mushrooms that have long been used as medicinal mushrooms such as Ganoderma lucidum, and edible mushrooms such as Tuber japonicum, a white truffle found in Japan. The extracts were tested for whitening activity at National Taiwan University. First, I tested the inhibitory activity of tyrosinase, an enzyme that helps producing melanin, a component that darkens the skin. As a result, seven types of mushrooms were found to have relatively high inhibitory activity. The seven mushrooms were tested for activity inhibition and toxicity using melanin-producing B16-F10 melanoma cells derived from mice. The results showed that six of the seven mushrooms (Ganoderma lucidum, Boletopsis grisea, Pleurotus ostreatus, Grifola frondosa, Albatrellus confluens, and Calvatia craniiformis) were found to have whitening activity in the non-toxic concentration range, and these six were tested using zebrafish. The results showed that all six extracts inhibited pigmentation without adverse effects on the organism. This could lead to the discovery of new, safe, whitening cosmetic materials of natural origin.