臺灣博碩士論文加值系統

螺旋藻（Arthrospira spp.） 又俗稱藍藻，為可食用藻類，含有豐富的蛋白質、必需氨基酸、必需脂肪酸、維生素及礦物質等營養素，並含有一種特殊之色素攜帶蛋白質—藻藍蛋白質（C-Phycocyanin, CPC），其具有抗氧化（antioxidation） 活性，可減少肝臟脂質過氧化，並有抗菌、抗腫瘤及免疫調節等功效，為理想之保健食品材料。有鑑於肝病為我國人主要疾病之一，本論文乃以螺旋藻為材料，研究其護肝（liver protection） 功效與作用之機制，期望能開發具市場價值之保健食品。本論文以四氯化碳（carbon tetrachloride, CCl4） 誘發大鼠形成慢性化學性肝損傷的模式，探討不同劑量之藻藍蛋白質對肝臟細胞之保護效果及其作用機制。結果顯示，在護肝功效評估的部分，經四氯化碳誘導之大鼠，投予藻藍蛋白質之組別與負對照組相較，其血清中麩氨酸氨基轉氨酶（aspartate aminotransferase, AST） 與麩氨酸丙酸轉氨酶（alanine aminotransferase, ALT） 兩者的上升幅度皆顯減著少（Ｐ＜0.05），顯示藻藍蛋白質可抑制四氯化碳引起之 AST 及 ALT 值上升。餵食藻藍蛋白質中、高劑量組之試驗大鼠，其肝臟中所含之抗氧化物質（GSH） 含量及抗氧化酵素（SOD、Catalase、和 GSH-Px） 活性，相較於負對照組亦明顯增加（Ｐ＜0.05）。病理切片結果顯示，餵食藻藍蛋白質可明顯改善肝臟空泡化變性、細胞腫脹、壞死、膽管增生及肝纖維化等異常病理變化之情形。綜合以上結果顯示，藻藍蛋白質具有提升肝臟所含之抗氧化物質與酵素含量、緩解四氯化碳所致之肝損傷、保護肝細胞完整性之效果。本論文結果顯示，由螺旋藻中萃取之藻藍蛋白質，對於開發護肝功效之保健食品具有正面意義。

Spirulina (Arthrospira spp., commonly known as cyanobacteria) is an edible alga, which is rich in proteins, essential amino acids, essential fatty acids, vitamins and minerals, and other nutrients. The Spirulina contains a special pigment carrying protein (C-phycocyanin, CPC). The CPC has effects on antioxidation, reduces liver lipid peroxidation, antibacteria, antitumor and immunoregulation, which could be ideal materials for health food. In this current paper, the effect of Spirulina on protection of liver and the mechanisms were studied to develop a health food for liver protection, since the liver disease is one of the main diseases in Chinese. The chronic chemical liver injury rat model was induced with carbon tetrachloride (CCl4). The effect of different doses of CPC protection effect and mechanism on liver cells were detected. The liver function analysis indicated that the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in sera of the model rats induced by CCl4 was significantly decreased (P<0.05) by the treatment of CPC comparing with that of the negative control rats. In liver tissues from the rats treated with high dose of CPC, the antioxidant enzymes, including SOD, Catalase, GSH and GSH-PX, were also increased significantly compared with that of the negative control group (P<0.05). The pathological observation showed that the pathological phenotype of cavitation of degeneration, liver cell swelling, necrosis, fibrosis were obviously improved by the treatment of CPC. In summary, the results indicated that the CPC treatment increased the levels of the antioxidant enzymes in liver and improved the liver function in the rat model of CCl4 induced chronic liver injury, and suggesting that antioxidant enzymes inducible activity of CPC could be one of mechanisms of liver protection. Our founding was also suggested that CPC could be a potential health food for liver protection.

摘 要 I
Abstract III
謝 誌 V
目錄 VI
圖表目錄 VIII
第一章 前言 1
第二章 文獻回顧 3
2.1 肝臟 3
2.1.1 肝臟組織結構與功能 3
2.1.2 肝臟疾病成因 8
2.1.3 肝纖維化機制 11
2.2 自由基與氧化壓力 12
2.2.1 自由基簡介 12
2.2.2 氧化壓力與細胞傷害 15
2.3 抗氧化物質 16
2.3.1 非酵素類抗氧化物質 16
2.3.2 酵素類抗氧化物質 17
2.4 四氯化碳誘導肝損傷 20
2.4.1 四氯化碳誘導肝損傷作用機制 20
2.4.2 四氯化碳於動物實驗之研究 23
2.5 螺旋藻簡介 25
2.5.1 螺旋藻之營養價值 26
2.5.2 螺旋藻之功效及應用 32
2.5.3 藻藍蛋白質結構與功能 34
第三章 論文研究 35
3.1 研究目的 35
3.2 材料與方法 36
3.2.1 實驗架構 36
3.2.2 藻藍蛋白質樣品製備 38
3.2.3 試驗動物 40
3.2.4 藥品與試劑 42
3.2.5 儀器與設備 43
3.2.6 試驗物質與對照物質的給予及試驗樣本收集 44
3.2.7 肝臟功能生化分析 45
3.2.8 抗氧化物質與酵素分析 46
3.2.9 組織切片及病理判讀 47
第四章 結果 51
4.1 攝食量、體重及臟器重量 51
4.2 臨床症狀觀察及病理切片分析 56
4.3 血清脂質生化值變化 65
4.4 血清白蛋白及總蛋白質值變化 70
4.5 肝纖維化指標、脂質過氧化程度分析 70
4.6 抗氧化物質與酵素分析 76
第五章 討論 78
5.1 對試驗動物體重、臟器相對重量及攝食量之影響 78
5.2 對血清生化值之影響 78
5.2.1 肝功能生化值 78
5.2.2 血清白蛋白及總蛋白質含量 79
5.2.3 血脂濃度 80
5.3 對肝纖維化之影響 80
5.4 對脂質過氧化程度之影響 81
5.5 對肝臟抗氧化物質含量與酵素活性之影響 82
第六章 結論 84
參考文獻 85
附錄 1 螺旋藻製劑之藻藍蛋白質含量測定結果 98
附錄 2 各成分定量標準曲線及決定係數（R2） 99
作者簡介 100

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