第一部分: 台灣苧麻葉部化學成分之研究 第二部分: 金童及金皇石斛水萃物對小鼠腸道菌相分布之影響

Abstract

本論文包含兩部分: 第一部分 : 台灣苧麻葉部化學成分之研究 台灣苧麻 (Boehmeria formosana) 分布於台灣、日本與中國大陸，為蕁麻科植物，同屬植物具有降血糖、抗B型肝炎、抗癌等生物活性，文獻指出其活性與生物鹼與原花青素相關。而本研究，相較於同屬植物，台灣苧麻葉部成分含有豐富的菲并吲哚啶(phenanthroindolizidine) 與菲并喹啉 (phenathroquinolizidine) 類生物鹼，此類生物鹼結構特色為三個苯環的菲 (phenanthrene) 接一個吲哚啶雙環的生物鹼。此類生物鹼於文獻中報導具有許多的生物活性，包含抗發炎、抗微生物、抗癌、抗痢疾、抗血管新生等，其中，對於抑制多種癌細胞生長之活性更為顯著。 取葉部乙醇粗提物以液相-液相分配進行極性劃分，分別為乙酸乙酯、正丁醇以及水可溶部分，而本研究大部分生物鹼分布在正丁醇層。利用Sephadex LH-20、CPC、半製備HPLC等層析方法，分離及鑑定出17個化合物: seco-Formonsanasine A (1), 4a,4b-seco-Dehydroantofine (2), seco-Formonsanasine B (3), Tylophoridicine D (4), Deoxytylophorinine (5), Boehmeriasin A (6), Boehmeriasine C (7), Tylophoridicine G (8), N-pyrrolidyl-3,4-dimethoxycinnamoylamide (9), 5,7-Dihydroxychromone-7-O-β-D-glucopyranoside (10), 5,7-Dihydroxychromone (11), Quercetin 3-O-α-L-rhamnoside (12), Quercetin (13), Hypoxanthine (14), 3,4- Dihydroxybenzoic acid (15), (2S,3S)-[3-(4'-methoxyphenyl)piperidin-2-yl]methanol (16)。其中，化合物 1、3、7、8為菲并吲哚啶類生物鹼，透過1D與2D NMR、MS (n)進行結構解析，透過文獻搜尋確認為新的化合物；化合物 9、16經由文獻搜尋結果得知為首次於天然物中分離得到。

第二部分 : 金童及金皇石斛水萃物對小鼠腸道菌的菌相分布之影響 石斛屬植物先前文獻指出其醣類成分具有許多生物活性，例如:抗氧化、抗糖尿病、抗腫瘤、神經保護作用等活性。蘭科石斛雜交種亦屬中藥材，具有生長快速的特性，相較於野生種石斛可能其有效成分較為豐富。 先前研究發現金童石斛粗萃多醣作用在胰臟β細胞，可對抗STZ的毒性，此研究顯示金童石斛粗萃多醣成分具有抗糖尿病功效的潛力。由於先前未曾報導其雜交種石斛與腸道菌的關聯性。因此本次實驗著重在初步了解C57BL/6J品系之實驗鼠食用此兩種石斛，為期7周餵養，期間紀錄生理數值 (體重、血糖、攝食量)，以及犧牲後取得脂肪組織的含量多寡，藉此作為初步的實驗依據。進而抽取犧牲當日殘留於直腸的糞便之DNA，利用PCR技術擴增16S V3-V4區域片段的DNA，以NGS分析其菌相分布情形。 本次實驗初步產出的結果進行菌相分析，初步探討菌屬與多醣成分可能的相關性，做為日後研究的基石。

Abstract This thesis are inclusive of two parts: Part 1: Chemical investigation on the leaves of Boehmeria formonsan Boehmeria formosana which belongs to Urticaceae is major distributed mainly in Taiwan, Japan and China. Its related species have been reported to possess bioactivities, including anti-glycemic effect, inhibition of HBV production, anti-cancer and so on. Some reports highlight that its bioactivities are arosed from cyanidins and alkaloids. In this study, B. formosana leaf contains aboundance phenanthroindolizidine-type and phenathroquinolizidine-type alkaloids in comparison with the same genus (Boehmeria). These chemical structure of alkaloids is characterized by three phenanthrene fuesd with an acridine bicyclic alkaloids. That alkaloids have many biological activities, including anti-inflammatory, anti-microbial, anti-cancer, anti-dysentery, anti-angiogenesis, etc. reported in the literature; especially, cytotoxic effect is more remarkable among above-mentioned bioactivities. The ethanol extract of B. formosana leaves were divided into ethyl acetate, n-butanol and water layer followed by liquid-liquid phase distribution. Most of the alkaloids in this study are distributed in the n-butanol layer. 16 compounds were isolated by Sephadex LH-20, CPC, semi-preparative HPLC including seco-Formonsanasine A (1), 4a,4b-seco-Dehydroantofine (2), seco-Formonsanasine B (3), Tylophoridicine D (4), Deoxytylophorinine (5), Boehmeriasin A (6), Boehmeriasine C (7), Tylophoridicine G (8), N-pyrrolidyl-3,4-dimethoxycinnamoylamide (9), 5,7-Dihydroxychromone-7-O-β-D-glucopyranoside (10), 5,7-Dihydroxychromone (11), Quercetin 3-O-α-L-rhamnoside (12), Quercetin (13), Hypoxanthine (14), 3,4- Dihydroxybenzoic acid (15), (2S,3S)-[3-(4'-methoxyphenyl)piperidin-2-yl]methanol (16). Among the isolated compounds, compounds 1, 3, 7, and 8 are phenanthroindolizidine-type alkaloids, which were confirmed to be new compounds based on 1D, 2D NMR and MS(n); compounds 9 and 16 were first isolated from the nature.

Part 2: Effects of dietary supplement with the water extracts from two Dendrobium hybrids (Cassiope and Taiseed Tosnobile) on the gut microbiome diversity in mice The previous literature of Dendrobium indicate that their carbohydrate components have many bioactivities, such as anti-oxidation, anti-diabetes, anti-tumor, neuroprotective effects. The hybrids of Dendrobium are Chinese herbal medicines, which have the characteristics of rapid growth and richer active constituents compared with wild-type Dendrobium species. In our previous studies showed that the crude polysaccharides from Dendrobium cassiope can counteract the toxicity of STZ in pancreatic β cells. This result showed that the polysaccharide extracted from Dendrobium cassiope has the potential to alleviate diabetes. Because it has not been reported previously that the relationship between Dendrobium species (Taiseed Tosnobile and Cassiope) and gut microbiota in mice. Therefore, in this study, we focused on the preliminary understanding of this relationship in mice after taking in Dendrobium species (Taiseed Tosnobile and Cassiope) for 7 weeks feeding. During the experimental term, the physiological values (weight, blood sugar, food intake) were recorded; the amount of tissue were obtained after sacrifice. The microbial gDNA of the feces remained in the rectum on the day of sacrifice was extracted, and DNA sequences of the 16S V3-V4 region were amplified by PCR. Next, the microbial distribution of the feces were analyzed by NGS. The results can not only be regarded as the basis for the further researches, but also firstly discussed the possible correlation between mictobiota and these 2 species of Dendrobium (Taiseed Tosnobile and Cassiope).