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研究生:林筱晴
研究生(外文):Hsiao-Ching Lin
論文名稱:樹豆對酒精誘導肝細胞損傷之保護效應 及其活性成分之研究
論文名稱(外文):Studies on the protective effect against alcohol-induced damage in liver cells and the bioactive components of Cajanus cajan L.
指導教授:吳思敬
指導教授(外文):She-Ching Wu
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:食品科學系研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
畢業學年度:100
語文別:中文
論文頁數:113
中文關鍵詞:樹豆抗氧化酒精性肝損傷花青素
外文關鍵詞:Cajanus cajanantioxidative activitiesalcohol-induced liver damageanthocyanins
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樹豆 [Cajanus cajan (Linn.) Millsp.] 為豆科 (Leguminosae) 木豆屬 (Cajanus) 之多年生矮灌木作物,具有調節血糖、抗菌、抗瘧疾等多種生理功效。本研究以樹豆臺東 3 號為原料,分析其主要生理活性成分,並探討樹豆對酒精誘導 Clone 9 大鼠正常肝細胞損傷之保護功效。樹豆種實以正己烷、乙酸乙酯、甲醇及 50% 乙醇等溶劑萃取,並以抗氧化能力作為篩選平台。結果顯示,50% 乙醇萃取物具最佳之清除 DPPH 自由基能力 (88.20%)、總抗氧化能力 (95.76%) 及還原力 (A700nm=1.51)。於酒精誘導肝細胞損傷方面,樹豆 50% 乙醇萃取物能有效減緩酒精誘導 Clone 9 細胞中三酸甘油酯蓄積及脂質過氧化程度,並可提升細胞中 glutathione (GSH) 含量。進一步分析樹豆之生理活性成分,使用 XAD-7 管柱層析及高效能液相層析進行分離及純化,純化之物質以液相層析串聯質譜儀 (LC-MS/MS) 進行結構鑑定。結果顯示,樹豆 50% 乙醇萃取物之區分層中,以 30% 甲醇區分層具有較佳之抗氧化能力,而其純化之物質經鑑定後確定結構為花青素 cyanidin-3-glucoside (C-3-G) 及 peonidin-3-glucoside (P-3-G)。而此區分物於酒精誘導肝損傷方面亦可有效減緩酒精所誘導之肝損傷,並可提升細胞中 catalase (CAT)、glutathione peroxidase (GPx)、glutathione reductase (GR) 及 glutathione-S-transferase (GST) 等抗氧化酵素之活性。綜合上述結果,樹豆 50% 乙醇萃取物及其 30% 甲醇區分物均具有預防酒精性肝損傷之功效,且主要活性成分為 C-3-G 及 P-3-G。
Pigeone pea [Cajanus cajan (Linn.) Millsp.] is one of the perennial shrub belongs to the Leguminosae family and genus of Cajanus. It is used for blood glucose regulation, antibacterial and antipaludism. In this study, the bioactive compounds of Cajanus cajan were determined and the potentiality of Cajanus cajan in protecting alcohol-induced damage of Clone 9 (rat liver cell) was evaluated. Cajanus cajan seeds was extracted by n-Hexane, ethyl acetate, methanol and 50% ethanol, then the antioxidant properties of the extracts were determined. The 50% ethanol extracts obtained from Cajanus cajan exhibited the strongest DPPH radical scavenging capacity (88.20%), total antioxidant activity (88.20%) and reducing power (A700nm=1.51). In the aspect of alcohol-induced liver cell damage, the 50% ethanol extracts obtained from Cajanus cajan not only significantly reduced triglyceride accumulation and lipid peroxidation but also enhanced the glutathione (GSH) content in the alcohol-induced Clone 9 cell. The 50% ethanol extracts obtained from Cajanus cajan was fractionated by Amberlite XAD-7 column. The bioactive components of Cajanus cajan were purified by HPLC. The chemical of purified materials were identified by LC-MS/MS. The results indicated that the 30% methanol fraction of the 50% ethanol extracts obtained from Cajanus cajan exhibited the best antioxidant activity and the purified compounds from this fraction were cyanide-3-glucoside (C-3-G) and peonidin-3-glucoside (P-3-G) confirmed by structure identification. This fraction could attenuate the alcohol-induced cell damage and enhance the activities of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST) in the alcohol-induced cell. These results suggest that the 50% ethanol extracts obtained from Cajanus cajan and its 30% methanol fraction exhibit the protective effect on the alcohol-induced liver cell and the major bioactive components are C-3-G and P-3-G.
中文摘要 I
英文摘要 III
謝誌 V
目錄 VI
圖次 VII
表次 X
前言 1
第一章、文獻回顧 3
一、樹豆 4
二、花青素 11
三、自由基與抗氧化防禦系統 17
四、肝臟生理及其傷害機制 24
研究目的 32
實驗架構 33
第二章、材料與方法 37
第三章、結果與討論 51
一、樹豆不同極性溶劑萃取物之萃取率 52
二、樹豆不同極性溶劑萃取物之抗氧化活性 52
三、樹豆 50% 乙醇萃取物預防酒精誘導 Clone 9 大鼠正常肝細胞損傷之能力 59
四、樹豆 50% 乙醇萃取物生理活性成分之分離純化及結構鑑定 69
五、樹豆 50% 乙醇萃取物之 30% 甲醇區分物 (F2) 預防酒精誘導 Clone 9 大鼠正常肝細胞損傷之能力 80
結論 98
參考文獻 99

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