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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/35102

Title: 海藻機能性脂質之鑑定及紫菜中含硫醣脂之保肝作用
Identification of Functional Lipids of Marine Macroalgae and Hepatoprotective Effect of Porphyra cripata SQDG
Authors: Cheng Jung Tsai
蔡政融
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 海藻;紫菜;不飽和脂肪酸;含硫醣脂;吳郭魚;硫代乙醯胺
Marine macroalgae;Porphyra crispata;Polyunsaturated fatty acids;Sulfoquinovosyldiacylglycerols;Tilapia (Oreochromis niloticus × Oreochromis aurea ♂);thioacetamide
Date: 2012
Issue Date: 2013-10-07T02:50:31Z
Abstract: 東北角海域二十一種海藻包含五種綠藻(裂片石蓴、滸苔、硬毛藻、束生剛毛藻和球松藻),十三種紅藻(皺葉紫菜、異色角叉菜、小杉藻、蜈蚣藻、硬盾果藻、海頭紅、唇孢藻、小珊瑚藻、傘房龍鬚菜、繩龍鬚菜、異枝軟骨凹頂藻、紅羽凹頂藻、縱胞藻)及三種褐藻 (厚葉馬尾藻、重緣葉馬尾藻和小海帶),脂質含量以藻體乾重計算,綠藻含15.36~20.15 mg / g、紅藻含18.57 ~28.34 mg / g 褐藻為13.11~19.56 mg / g;脂質之多元不飽和脂肪酸 (polyunsaturated fatty acids, PUFAs) 含量以綠藻較高的為C18:2 n6及 C18:3 n3、紅藻為C20:4 n6 及 C20:5 n3、褐藻為 C18:4n3、C20:4n6 及C20:5n3;紅藻之 PUFAs 含量最高,以ecicosapentaenoic acid (EPA) 含量最為豐富,又以紫菜 (Porphyra spp.) 之 PUFA 及 EPA 含量最高,各藻種中均含有陸上植物缺乏之奇數碳脂肪酸 C17:0 和 C17:1,可作為分辨植物油及藻油之參考。 紫菜 (Porphyra crispata) 經冷凍乾燥及磨粉 (200 mesh) 後,以無水酒精萃取,紫菜酒精萃取物經 Diaion HP-20 樹脂分離後,得glycolipids,再以弱陰離子交換纖維素 (DEAE-cellulose, diethylaminoethyl-cellulose) 分離,經冰丙酮沉澱後,得 SQDG (sulfoquinovosyldiacylglycerol) 純品 ,收率為 0.032 % dry basis。紫菜之 SQDG 主要含 palmitic acid (C16:0, 33.3%)、EPA (C20:5n-3, 30.0%)、arachidonic acid (C20:4n-6, 12.7%)、oleic acid (C18:1n-9, 7.52%) 和 stearic acid (C18:0, 6.83%) 等脂肪酸,不飽和脂肪酸n - 3 / n - 6 比值為 1.9,其抑制人類肝癌腫瘤細胞 (Hep G2) 之半抑制濃度 (IC50 ) 為126 μg /mL,低於菠菜SQDG (255 μg / mL)。 為評估紫菜及其酒精萃取物於體內之保肝功效,故建立吳郭魚 (Oreochromis niloticus × Oreochromis aurea ♂) 肝損傷之動物模式。吳郭魚每週三次以腹腔注射 thioacetamide (TAA) 100 mg / kg b.w. 誘導肝損傷,持續 12 週後,其肝體比 (hepatosomatic index, HSI) 顯著 (P<0.05) 增加,肝組織可明顯觀察到肝細胞受損,導致血清中 AST (2.6 倍) 與 ALT (2.8倍) 活性上升、紅血球之形態改變,形成棘狀細胞 (burr cells) ,因此 TAA 可誘發吳郭魚肝損傷。應用此肝損傷動物模式,評估紫菜粉 (P) 、紫菜酒精萃出物 (PE) 及紫菜SQDG之保肝功效,吳郭魚以 TAA 誘導肝損傷,同時餵食含 1%、5% 紫菜粉 (1% P、5% P) 及0.1%、0.5% 紫菜酒精萃出物 (0.1% PE、0.5 % PE) 和紫菜 SQDG (20 mg/kg day-1) 之飼料,結果 5% P、 0.1% PE、 0.5% PE 及紫菜 SQDG組 均有保護肝損傷的功效,其中又以 0.5% PE 及 SQDG 組別於 TAA 誘導肝損傷過程中可顯著降低血清中 AST 及ALT 之活性,故以此吳郭魚肝損傷動物模式可用於評估健康食品是否具有保肝作用,紫菜之酒精萃取物及SQDG 具有開發為保肝健康食品之潛力。
The fatty acid compositions of 21 species of marine macroalgae, including 5 species of Chlorophyta (green algae), 13 species of Rhodophyta (red algae) and 3 species of Heterokontophyta (brown algae) were collected from the Northeastern Taiwan to survey their functional lipids. The lipid contents of green algae ranged 15.36~20.15 mg/g dry basis (db) were characterized by high content of C18:2 and C18:3; red algae (18.57 ~28.34 mg / g db) were high in C20:4 and C20:5; while brown algae (13.11~19.56 mg / g db) were high in C18:4, C20:4, and C:20:5. All algal lipids contained fatty acids of odd-number carbon, C17:0, and C17:1. Red algae had relatively higher levels of polyunsaturated fatty acids (PUFAs) and richer in ecicosapentaenoic acid (EPA) than green and brown algae. Red alga, Porphyra crispata was extracted with ethanol (PE). The extract consisted of PE was separated by hydrophobic column (Diaion HP-20 column) to obtain glycolipid fraction. Sulfoquinovosylglycolipid (SQDG) was purified from glycolipid fraction using DEAE-cellulose column chromatography. The main fatty acids in SQDGs were palmitic acid (C16:0) 33.3%, eicosapentaenoic acids (EPA, C20:5, n-3), 30.0 %, arachidonic acid (C20:4, 12.7%), oleic acid (C18:1), 7.52% and stearic acid (C18:0), 6.83 %. The n - 3/n - 6 ratio was 1.9 while the authentic standard, spinach SQDG did not contain n-3 fatty acids. Sulfoglycolipids inhibited the growth of human hepatocellular carcinoma cell line (Hep G2). The IC50 was 126 μg /mL being lower than the spinach SQDG (255 μg / mL). The SQDGs from Porphyra crispate has shown in vitro cytotoxicity against human hepatoma (HepG2) cells that was further studied for its in vivo hepatoprotective activity. For this reason, the aim of this study was to establish an animal model of chronic liver damage and to investigate the in vivo hepatoprotective effect of the ethanol extract of Porphyra crispata and its sulfoglycolipid fraction. Tilapia (Oreochromis niloticus × Oreochromis aurea ♂) was used as animal model to study liver injury through intraperitoneal administration of thioacetamide (TAA), and hepatprotective effect of the freeze – dried powder of Porphyra crispate (P), PE and SQDG. The results showed that TAA administration (100 mg / kg i.p., per 2 day) for 12 weeks caused liver damage indicated by significant (P<0.05) increase in hepatosomatic index (HSI), serum AST (2.6 times) and ALT (2.8 times), and a significant (P < 0.05) decrease in triacylglycerol and total cholesterol levels and liver section compared to those without TAA (saline - 0P). Tilapia (Oreochromis niloticus) challenged with TAA showed abnormal red blood cells (burr cells). The occurrence of burr cells decreased when tilapia were fed 0.1%、0.5 % PE. In order to compare the hepatoprotective effect of 1%, 5% P, 0.1%, 0.5 %PE and SQDG (20 mg/kg day-1) with the challenged and the unchallenged control, PE and SQDG showed potential hepatoprotective effects intilapia fish model.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0D93320008
http://ntour.ntou.edu.tw/handle/987654321/35102
Appears in Collections:[食品科學系] 博碩士論文

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