臺灣博碩士論文加值系統

本研究取綠藻水萃物進行抗發炎之相關試驗，結果顯示，綠藻水萃物可以有效抑制LPS誘發RAW264.7巨噬細胞iNOS和COX-2蛋白之表現。
另外為了解綠藻水萃物延緩衰老之實際功效，本試驗採隨機、安慰劑對照之人體臨床試驗設計，共計44位健康受試者全程參與（男性11人，女性33人），分為兩組同步進行，實驗組之受試者每天飲用綠藻水萃物27mL；對照組之受試者每天飲用安慰劑27mL。試驗期間共計180天，試驗期之第0、90、180天，進行體位測量並收集血液檢體進行試驗，且於試驗其之第0及180天進行腹部超音波檢測。
結果顯示，飲用綠藻水萃物180天，可以顯著增加血漿中總抗氧化能力、維生素C、麩胱甘肽、多酚化合物、含硫功能基含量，及顯著降低血漿中8羥基-2’-去氧鳥嘌呤核甘及降低脂質之過氧化反應，並且顯著提高血球中葡萄糖六磷酸去氫酶、觸酶、麩胱甘肽過氧化酶、麩胱甘肽還原酶及超氧歧化酶等抗氧化酵素之活性。
此外，由腹部超音波檢測結果顯示，服用綠藻水萃物可顯著改善受測者脂肪肝、膽結石和 肝實質性病變之效果。

In this study, Chlorella water extracts was used for determining anti-inflammation. Results showed that Chlorella water extracts inhibited iNOS and COX-2 protein expression on LPS-induced RAW 264.7 macrophage.
To further evaluate the anti-aging effect of Chlorella water extracts in human. A randomized and placebo-controlled study was designed. Forty-four subjects (11 males and 33 females) were completed involved in this study. The total experimental period was 180 days. One group of subjects took placebo, and Chlorella water extracts for the other one group subjects. Each subject took 27mL placebo or Chlorella water extracts every day for continuous 180 days. Anthropometric measurements and blood collection were performed at the initial, 90, and 180 days of the testing period. Abdominal ultrasound was performed at the initial, and 180 days. Results showed that 180 days of continuous administration of Chlorella water extracts can significantly increase total antioxidant capacity, ascorbic acid, glutathione (GSH ), total phenolics, and total thiol groups, but reduce 8-hydroxydeoxyguanosine (8-OH-dG), and TBARS contents in plasma. The activities of glucose-6-phosphate dehydrogenase (G-6-PDH), catalase (CAT), glutathione reductase (GSH Rd), glutathione peroxidase (GSH Px), and superoxide dismutase (SOD) in erythrocytes were also greatly increased.
In addition, the abdominal ultrasound examination showed that Chlorella water extracts could improvement on fatty liver, gall stone and parenchyma liver disease.

表次......................................................I
圖次.....................................................Ⅲ
附錄.....................................................IV
中文摘要.................................................V
英文摘要.................................................VI
壹、 前言...............................................1
貳、 文獻整理...........................................2
一、 綠藻的分類........................................ 2
二、 綠藻.............................................. 4
三、 綠藻的營養成分.................................... 4
四、 綠藻之保健功效.................................... 6
(一) 提升抗氧化能力...................................6
(二) 調節血脂.........................................7
(三) 調節免疫.........................................8
(四) 排除毒素.........................................9
五、 發炎反應..........................................9
(一) 一氧化氮及誘導型一氧化碳合成酶..................9
(二) 前列腺素E2及第二型環氧化酶.....................10
六、 自由基、氧化壓力與老化理論........................13
(一) 老化的定義......................................13
(二) 老化與自由基....................................14
(三) 老化與抗氧化防禦系統............................17
(四) DHEA/DHEAS與老化.............................21
(五) 8羥基-2’-去氧鳥嘌呤核甘與老化...................24
(六) 發炎與老化......................................26
參、 研究目的..........................................28
肆、 試驗設計..........................................29
伍、 材料與方法........................................30
一、 試驗材料..........................................30
(一) 綠藻水萃物......................................30
(二) 人體試驗........................................31
1. 基本資料......................................33
2. 血液樣品收集..................................33
二、 試驗用藥.........................................36
三、 試驗儀器..........................................41
四、 試驗方法..........................................43
(一) 綠藻水萃物酚類化合物含量測定............. .....43
1. 總酚含量.......................................43
(二) 巨噬細胞RAW264.7培養.........................43
1. PBS製備.......................................43
2. RPMI 1640培養液製備...........................43
3. 冷凍細胞活化...................................44
4. 細胞培養.......................................44
5. 細胞冷凍保存...................................44
6. 細胞計數.......................................45
(三) 抗發炎性測定..................................46
1. 細胞存活試驗...................................46
2. iNOS與COX-2測定.............................47
(四) 人體臨床試驗..................................49
1. 血液樣本之收集................................49
2. 血液生化值分析................................49
3. 肝功能評估....................................50
4. 血漿抗氧化活性之測定..........................51
(1) 血漿中總抗氧化狀態測定....................51
(2) 血漿中脂質過氧化物含量測定...............51
(3) 血漿中含硫功能基之測定...................52 (4) 血漿中麩胱甘肽（GSH）之測定..............52
(5) 血漿中總酚類化合物含量測定...............53
(6) 血漿中維生素C含量測定...................53
(7) 血漿中8羥基-2’-去氧鳥嘌呤核甘（8-OH-dG）之高效率層析法之測定.......................54
5. 抗氧化酵素活性分析............................56
(1) 紅血球超氧歧化酶(superoxide dismutase；SOD)活性測定......................................56
(2) 紅血球葡萄糖六磷酸去氫酶(glucose-6-phosphate dehydrogenase；G6PD)活性測定...............58
(3) 紅血球麩胱甘肽過氧化酶(glutathione peroxidase；
GSH Px)活性測定..........................59
(4) 紅血球麩胱甘肽還原酶(glutathione reductase；
GSH Rd)活性測定..........................60
(5) 紅血球觸酶活性測定（catalase）..............61
6. 自由基測定....................................62
(1) 全血超氧陰離子............................62
五、 統計分析...........................................63
陸、 結果與討論........................................64
一、 酚類化合物分析...................................64
（一） 綠藻水萃物總酚含量測定.........................64
二、 抗發炎試驗.......................................64
(一) 對巨噬細胞RAW264.7存活率之影響...............64
(二) 對巨噬細胞RAW264.7中iNOS表現之影響..........64
(三) 對巨噬細胞RAW264.7中COX-2表現之影響........64
三、 人體試驗.........................................70
(一) 受試者特性....................................70
(二) 對受測者血液生化值的影響......................70
(三) 全血自由基影響................................74
(四) 對受測者血漿抗氧化活性之影響..................76
(五) 對受測者紅血球中抗氧化酵素活性之影響..........87
(六) 肝功能評估....................................95
柒、 結論.............................................100
捌、 參考文獻.........................................101
玖、 附錄.............................................109

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