臺灣博碩士論文加值系統

檄樹(Morinda citrifolia L.)又名諾麗(noni)，長久以來為夏威夷及大溪地島民之民俗藥物。近年來熟成諾麗果汁紛紛以直銷形式進口，宣稱具有抗氧化、抗癌與促進免疫等健康功效。然而其中僅有少數功效經科學驗證。製造傳統熟成果汁耗費時間長，果汁中的有效成份也隨熟成過程逐漸減少。故本研究探討輔以商業化果膠酵素新鮮諾麗果汁之製造，及其不同儲藏溫度(4 °C或室溫)與避光條件，一個月期間之化學與物理變化。並探討Palawan商業化熟成果汁及其乙醇萃取物，對食道癌細胞(CE 81T)與結腸癌細胞(HTB-37)之抗癌作用。結果顯示果膠酵素能夠澄清果汁、增加果汁的產量，增加東茛萣素(scopoletin)的含量。新鮮果汁經儲藏後其總抗氧化能力(TEAC) 與總酚含量下降、東茛萣素與縮合單寧含量稍微增加。且縮合單寧含量與紅色度值(a* value)有正相關。熟成諾麗果汁及其酒精萃取物於作用八小時兩細胞株之DNA均輕微損傷；二十四小時HTB-37凋亡、CE 81T則呈現壞死；七十二小時後導致兩株人類腫瘤細胞死亡。細胞的死亡與其作用劑量呈正相關。以不同抗氧化模式分析熟成諾麗果汁及其酒精萃取物，顯示兩者各擁有不同之抗氧化能力。綜合而言，諾麗果汁及其酒精萃取物是先造成細胞DNA損傷、使細胞壞死或凋亡。

Morinda citrifolia L., also known as noni, has a long history of traditional use in the Hawaiian and Tahitian islands as cure-all medicine. Recently, commercial fermented noni juice products have been imported from different countries and delivered directly to consumers with claims of multiple health benefits, including antioxidative, anticancer and immuno-stimulating activities. Only few of its health benefits have been confirmed by scientific reports. Production of noni juice by fermentation is time consuming and some of its active ingredients will decompose during its fermentation process. In this study, we investigate the production of fresh noni juice enhanced by commercial pectinase and change in its chemical and physical properties during one-month storage at 4 °C (refrigerator) or room temperature with or without exposed to light. Palawan, a commercial fermented noni juice, and its ethanol extracts were chosen to test their anticancer effects on Carcinoma of Human Esophagus cell (CE 81T) and Human colon cancer cell (HTB-37) in cell culture. The results indicated that addition of pectinase would enhance the clarification of noni juice and increase juice production and scopoletin content. During storage, Trolox equivalent antioxidant capacity (TEAC) and total phenol content of noni juice were found to decrease, whereas scopoletin and condensed tannin contents were found to increase slightly. Moreover, red color index (a* value) showed positively correlated to condensed tannin. Fermented noni juice and its ethanol extracts were found to cause both slightly DNA damage in 8hr; HTB-37 apoptosis in 24 hr, but necrosis in CE 81T and both cell lines death in 72 hr. And the death is dose-dependent. Noni juice and its ethanol extracts possessed different antioxidative activities in different antioxidant assay model. Taken together, our observations suggest that noni juice and its ethanol extracts induce DNA lesion eventually cell death.

目錄----------------------------------------------------------------------------------------------I
表次----------------------------------------------------------------------------------------------III
圖次----------------------------------------------------------------------------------------------IV
附錄次--------------------------------------------------------------------------------------------V
中文摘要----------------------------------------------------------------------------------------VI
英文摘要---------------------------------------------------------------------------------------VII
壹、前言-----------------------------------------------------------------------------------------1
貳、文獻回顧-----------------------------------------------------------------------------------xx
一、簡介------------------------------------------------------------------------------------2
二、諾麗果汁之加工製造---------------------------------------------------------------4
三、生理功效------------------------------------------------------------------------------5
參、材料與方法-------------------------------------------------------------------------------13
一、諾麗果汁與諾麗鮮果---------------------------------------------------------------13
二、試劑與試藥---------------------------------------------------------------------------13
三、儀器設備------------------------------------------------------------------------------14
四、其他------------------------------------------------------------------------------------14
五、添加酵素生產諾麗果汁------------------------------------------------------------15
（一）果實前處理與添加酵素生產果汁方式----------------------------------15
（二）分析果汁固形物與東茛萣素(scopoletin)含量--------------------------16
（三）果汁顏色變化、TEAC抗氧化能力、總酚含量、縮合單寧含量與scopoletin含量之儲藏試驗---------------------------------------------------------16
A. 顏色變化-------------------------------------------------------------------16
B. 總抗氧化能力(Trolox equivalent antioxidant capacity, TEAC) ---17
C. 總酚含量-------------------------------------------------------------------17
D. 縮合單寧含量-------------------------------------------------------------18
E. 測定scopoletin含量--------------------------------------------------------18
六、以乙醇分離熟成果汁進行細胞實驗--------------------------------------------------19
七、培養細胞之條件--------------------------------------------------------------------------20
（一）細胞培養基配製--------------------------------------------------------------20
（二）解凍細胞-----------------------------------------------------------------------20
（三）繼代細胞-----------------------------------------------------------------------20
（四）凍存細胞-----------------------------------------------------------------------21
八、細胞相關試驗-----------------------------------------------------------------------------21
（一）測定細胞存活率(XTT assay) ----------------------------------------------21
（二）使用流式細胞儀測定細胞凋亡--------------------------------------------21
（三）慧星試驗(comet assay) ------------------------------------------------------23
九、測定抗氧化能力--------------------------------------------------------------------------24
（一）總抗氧化能力(Trolox equivalent antioxidant capacity, TEAC) -------24
（二）清除α,α-diphenyl-β-picrylhydrazyl (DPPH)自由基能力-----------24
（三）過氧化氫清除能力-----------------------------------------------------------25
（四）還原力--------------------------------------------------------------------------25
（五）螯合亞鐵離子能力-----------------------------------------------------------26
十、功能性成分測定--------------------------------------------------------------------------26
（一）總酚含量-----------------------------------------------------------------------26
（二）縮合單寧含量-----------------------------------------------------------------26
（三）測定scopoletin含量------------------------------------------------------------26

肆、結果與討論---------------------------------------------------------------------------------27
一、新鮮諾麗果汁之加工----------------------------------------------------------------27
二、新鮮諾麗果汁之儲存試驗----------------------------------------------------------31
三、熟成諾麗過果汁抑制癌細胞相關試驗-------------------------------------------40
四、熟成諾麗果汁抗氧化性質分析----------------------------------------------------48
伍、結論--------------------------------------------------------------------------------------53
陸、參考文獻--------------------------------------------------------------------------------55
柒、附錄--------------------------------------------------------------------------------------59

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