中文摘要 三裂葉蟛蜞菊(Wedelia trilobata (L.) Hitchc.) 又稱南美蟛蜞菊, 為菊科多年生草本植物,常應用於美化環境,亦是坊間青草茶常用之 原料,具有清熱解毒,抗菌及抗腫瘤作用。本研究以三裂葉蟛蜞菊的 花 (flower) 、葉 (Leave)、莖 (stem) 為原料,分別測試其抗氧化 力、抑菌作用、抗發炎能力與抗肝癌效用。 三裂葉蟛蜞菊花、葉、莖三部位分別以乙醇、正己烷、乙酸乙酯、 正丁醇及蒸餾水等不同極性之溶劑分層萃取,再進行各層的抗氧化 力、抑菌能力及最小抑菌濃度試驗。另外小鼠巨噬細胞(RAW264.7) 細胞株試驗,以脂多醣體(Lipopolysaccharide,LPS) 促使其發炎反 應,分別以花、葉、莖之乙醇粗萃物以及水萃物來測試是否具有抑制 發炎介質的能力,再將效果最佳之花以不同極性之溶劑分層萃取測試 其活化 PPARγ 及PPARα 之能力。在肝癌 (SK-Hep-1) 細胞株的抑制 生長能力試驗方面則分別以乾燥及新鮮的三裂葉蟛蜞菊來測試,本實 驗採測試細胞存活率 (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide,MTT assay) 來觀察其生長情形,探討各樣本是否具有抑制癌細胞生長效果; 而抑制轉移試驗,則以測定肝癌細胞分泌matrix metalloproteinase (MMP)-9 及MMP-2 侵襲因子表現及侵入能力,來測試蟛蜞菊否具 有抑制SK-Hep-1 細胞轉移效果。 實驗結果得知三裂葉蟛蜞菊各萃取層都具有抗氧化能力,其中以 花及莖乙酸乙酯層能力最佳其總酚含量亦較多;而螯合亞鐵離子能力 則是以正丁醇層較佳。抑菌試驗結果發現,除了水層外,其對於仙人 掌桿菌 (Bacillus cereus) 和金黃色葡萄球菌 (Staphylococcus aureus) 都有顯著的抑制作用,而以乙酸乙酯層效果較佳,其對仙人掌桿菌最 小抑菌濃度可低達6.25 mg/mL。抗發炎介質NO2 方面,以花有較佳 的抑制效果。而花之不同極性層對PPARγ 則無活化效果,但花之乙 醇粗萃物及正己烷層具有很好的活化PPARα 之能力。抗肝癌試驗發 現,花之乙醇層具有最佳抑制肝癌生長能力,而花水煮20 分鐘之凍 乾物亦具有抑制能力,唯其效果明顯低於乙醇層。在細胞轉移試驗結 果發現,在抑制癌細胞分泌侵襲因子表現上以花的乙醇萃取物有較 佳,能降低16.82±2.04% MMP-9 表現量及12.45±2.28% MMP-2 表現 量,在侵入性實驗則以乙醇萃取液濃度80 μg/mL 有最好效果。綜合 上述,三裂葉蟛蜞菊其花部位具有較多的有生理活性物質,且證實了 花乙醇萃取物有抗肝癌細胞之轉移作用及抑制其生長的效果。
Abstract Wedelia trilobata (L.) Hitchc. is used in folk medicine. It is reported that the plant shows antibacterial activity and contents antitumor chemical components. The purpose of this research was to evaluate antibactes, antioxidation, anti-inflammation, and anticancer activity of W. trilobata. W. trilobata flowers , leaves and stems were extracted with ten times of ethyl alcohol. The extract was then partitioned by n-Hexane, ethyl acetate, n-butyl alcohol and water to evaluate its antimicrobial and antioxidative activities. We also evaluate its anti-inflammation and anticancer activity of the flower. The result showed that most extracts had antimicrobial activities except the water extracts from flower. The ethyl acetate extract was the most effective among all the extracts. Most extracts from flowers, leaves and stems showed the good antioxidative activities especially the flower. The the ethyl acetate extracts presented the best on DPPH scavenging activity and reducing power and had the highest total phenolic compounds content . We used the ethyl alcohol and water extracts of W. trilobata to treat RAW264.7 macrophage, which induced inflammation by LPS. In the NO2 assay, the extracts of W. trilobata flower had better inhibitory ability .In transient transfection assay shows the n-hexane and ethyl alcohol extracts of W. trilobata flower can activate PPARα. In MTT assay of SK-Hep-1, we found that extract of W. trilobata flower had the best inhibitory ability. We also found the ethyl alcohol extract of W. trilobata had the best ability to diminish the expression of matrix metalloproteinase (MMP)-9 and MMP-2. The ethyl alcohol extracts of flower had good anti-migration and anti-invasion ability especially on 80 μg/mL dose.