大安水蓑衣成分及相關活性探討

The constituents and their related activities of Hygrophila pogonocalyx Hayata

林建宏(Chien-Hung Lin)

指導教授 ： 李美賢

共同指導教授 ： 李宗徽(Tzong-Huei Lee)

臺北醫學大學/藥學院/生藥學研究所/碩士(2009年)

摘要

大安水蓑衣（Hygrophila pogonocalyx Hayata）為爵床科水蓑衣屬（Hygrophila）多年生水生挺水植物，為台灣特有種，產於台中縣臨海鄉鎮。根據本實驗室先前實驗結果顯示，大安水蓑衣95％乙醇萃取物，具清除自由基的活性（HO•、O2•‾及ABTS•‾，IC50 = 0.78、12.75及 7.94 ug/mL），因其被國際自然及自然資源保育聯盟 (IUCN)，列為瀕臨絕滅級 (endangered )，故本研究將組織培養的大安水蓑衣地上部以 95％ 乙醇萃取，並進行成分分離及其活性探討。經管柱層析分離純化得到十三個化合物，由物理和光譜相關數據解析後，分別為flavones 結構 luteolin 7-O-β-D-glucopyranoside（1），luteolin 7-O-β- D-glucuronide（2）；flavonols 結構 quercetin（3），isoquercitrin（4），rutin（5），myricetrin（6）；phenylethanoid glycosides 結構 acteoside（7），isoacteoside（8），β-ethoxylacteoside（9）；alkylated glycoside 結構 3-O-[β-D-apiofuranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol（10），3-O-[β-L-xylopyranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol（11）；及 steroid 結構 β-sitosterol（12）及 stigmasterol（13），以上十三個化合物均首次由大安水蓑衣中分離得到。此十三個化合物，除化合物 3 及 4 因量少、化合物 12 及 13 因為混合物未測試外，化合物1、2、5 ~ 11 進行神經生長因子分化的 PC12 細胞活性篩選試驗，在細胞存活率測試中，於100 μM 濃度下，發現九種化合物的細胞存活率皆大於 85%，進一步進行以六-羥基多巴胺誘導神經生長因子分化的 PC12 細胞毒性模式，結果顯示化合物 luteolin 7-O-β-D-glucopyranoside（1）具最佳細胞保護活性。十三個化合物，除化合物 12 及 13 因為混合物未測試外，化合物 1 ~ 11 進行人類黑色素細胞內酪氨酸酵素活性試驗，在細胞存活率測試中，於100 μM 濃度下，發現十一種化合物的細胞存活率皆大於 85%，進一步對人類黑色素細胞內酪氨酸抑制活性測試，結果顯示化合物 acteoside（7） 具有較佳抑制酪氨酸酵素活性效果。

關鍵字

大安水蓑衣 ； 臺灣特有種 ； 神經細胞保護 ； 人類黑色素細胞 ； 酪氨酸酵素

並列摘要

Hygrophila pogonocalyx Hayata is an endemic species in Taiwan. It was found along the seashore in Taichung country and was endangered according to the Red List Categories of the International Union for Conservation of Nature and Natural Resources (IUCN). Our previous results showed that the 95％ ethanol extract of Hygrophila pogonocalyx exhibited free radical scavenging activities (HO•, O2•‾ and ABTS•‾, IC50 = 0.78, 12.75 and 7.94 ?慊/mL). Therefore, the active constituents of H.pogonocalyx from tissue cultures (providing by Dr. Chin-Wen Ho, Tatung University, Taipei) are isolated in the present study. The structures of isolated compounds were identified by various physical and spectroscopic characterizations (eg. ESI-MS, UV, IR and NMR etc.). Thirteen compounds from Hygrophila pogonocalyx Hayata were identified in this study, including two flavones, luteolin 7-O-β-D-glucopyranoside (1) and luteolin 7- O-β-D-glucuronide (2); four flavonols, quercetin (3), isoquercitrin (4), rutin (5) and myricetrin (6); three phenylethanoid glycosides, acteoside (7), isoacteoside (8) and β-ethoxyl-acteoside (9); two alkylated glycosides, 3-O-[β-D- apiofuranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol (10) and 3-O- [β-L-xylo- pyranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol (11) and two steroid, β-sitosterol (12) and stigmasterol (13). Those 9 compounds were further evaluated the neurocytoprotective activity in NGF-differentiated PC12 cells, and compound 1 shows the most potent activity. Those 11 compounds were also evaluated the inhibition of cellular tyrosinase activities in human epidermal melanocytes（HEMn）. Cell viabilities of compounds 1 ~ 11 were higher than 85％, the compound 7 was the most potent in inhibiting tyrosinase activity.

並列關鍵字

Hygrophila pogonocalyx Hayata ； endemic species in Taiwan ； nerve growth factor-differentiated PC12 cell ； human epidermal melanocytes

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