臺灣博碩士論文加值系統

蔬果發酵纖維是由100種天然蔬菜及果實經酵母菌、乳酸菌等益生菌進行為期兩年的發酵，萃取、過濾『酵素』後所產生之蔬果纖維。為有效開發及利用此纖維殘渣，本研究利用體外、動物及人體實驗分別探討蔬果發酵提煉後的植物纖維是否具有益菌功能，是否可改善宿主腸道菌相及增進宿主免疫功能。
利用體外實驗評估『蔬果發酵纖維』和已知的『益菌生』菊醣或寡多醣對四種益生菌之益菌功效，實驗結果顯示『蔬果發酵纖維』具有益菌之特性，其中對乳酸桿菌的益菌效果較比菲德菌佳。
人體實驗共17位健康受試者參與，每位受試者均參與五階段、為期十週的試驗：第一階段為一週適應期，第二階段為三週實驗I期，受試者每日服用十顆不含乳酸菌之蔬果發酵纖維錠，第三階段為二週排空期，第四階段為三週實驗II期，受試者每日服用十顆添加LGG乳酸桿菌之蔬果發酵纖維錠，最後為一週排空期。結果發現服用蔬果發酵纖維錠後可有效增加宿主腸道中乳酸桿菌和比菲德菌等益生菌數，但對抑制有害菌指標產氣夾膜梭狀桿菌的能力則不如服用添加LGG乳酸桿菌之蔬果發酵纖維錠組。本實驗證明每日服用添加LGG乳酸桿菌之蔬果發酵纖維之『共生菌』具有最佳之腸道保健功效，但在停止服用『共生菌』進入排空期後有害菌又逐漸增加。
在動物實驗中，我們以BALB/c小鼠為實驗動物，將其分為二組，每組12隻。參考人體服用劑量，每日提供每隻小鼠1粒400 mg之蔬果發酵纖維錠或添加LGG乳酸桿菌之蔬果發酵纖維錠，實驗為期八週。根據小鼠腸道菌相及免疫功能之檢測結果，再次證明每日服用蔬果發酵纖維錠或『共生菌』二星期後小鼠腸道中益生菌明顯增加，有害菌指標大腸桿菌生長相對受到抑制，具改善腸道菌相的保健功效，但此結果至第五週已至停滯期 (plateau)，將實驗期延長為八週，仍未能讓宿主腸道中的益生菌持續增加。在動物實驗免疫評估的部分，服用添加LGG乳酸桿菌之蔬果發酵纖維錠兩週後，顆粒球及單核球吞噬能力明顯較蔬果發酵纖維錠組增加，此結果可能與BALB/c小鼠腸道中LGG乳酸桿菌快速增加有關。服用『共生菌』組小鼠脾臟細胞分泌IFN-γ的量比蔬果發酵纖維錠組高，而IL-10的分泌量則下降，此結果與文獻報告LGG乳酸桿菌可增進 Th1免疫調節功能之結果一致。
綜合評估：蔬果發酵纖維可以促進腸道中益生菌生長並抑制有害菌繁殖，故可有效利用此蔬果發酵纖維殘渣，添加LGG乳酸桿菌，進一步發展為具有改善宿主腸道菌相及免疫調節等保健功效之『共生菌』。

“Koso” is a healthy supplement extracted from 100 kinds of vegetables and fruits fermented by yeast and the lactic acid bacteria for two years. After “Koso” extraction, numerous fermented plant fibers were discarded. In order to reuse these fermented plant fibers efficiently, the aim of this study was to explore whether they may improve the host’ s gut microflora and immunity by in vitro and in vivo（human and animal ）experiments.
Comparing to the well-known prebiotics fructo-oligosaccharide or inulin, the fermented plant fibers had better prebiotic function to lactobacilli than bifidobacteria by in vitro assay.
Seventeen healthy volunteers (age: 18~35 years) were recruited into this free-living and self control study. The 10-week trial comprised an initial lead-in week, a 3-week treatment I phase (taking fermented plant fibers alone, ten tablets once a day), a 2-week washout phase, a 3-week treatment II phase (taking fermented plant fiber with L. rhamnosus GG), and followed by a final week of washout. After oral administration of the fermented plant fibers for 3 weeks, the fecal lactobacilli and bifidobacteria were increased efficiently. However, the decrease of fecal harmful bacteria, Clostridium perfringens in fermented plant fibers group was inferior to that of the synbiotic group. In this study we demonstrated that consumption both of the fermented plant fiber and L. rhamnosus GG may have the best effect on our intestinal health. However, the harmful bacteria may reappear in our intestine after stopping administration of the symbiotics.
Twenty four BALB/c mice were divided into two groups (prebiotics and synbiotics groups). Referring to the human dosage, each mouse was daily fed with 400mg fermented plant fiber or fermented plant fiber with L. rhamnosus GG for 8 weeks. In this animal study, we also proved that taking fermented plant fibers alone or fermented plant fibers with L. rhamnosus GG for two weeks could increase the fecal lactobacilli and bifidobacteria, while decrease the fecal coliform bacteria. However the increase of fecal beneficial bacteria may reach to a plateau after 5 weeks, even the feeding period was extended to eight weeks. For investigating the immunomodulating effect of the fermented plant fibers on BALB/c mice, blood samples were drawn from the retroorbital sinus every two weeks, and the mice were sacrificed on day 56. The phagocytosis of the neutrophils and monocytes in the synbiotic groups were significantly better than those of the prebiotics group. These results might be correlated with the increasing of L. rhamnosus GG in the intestine of the synbiotic groups. Compared the splenocyte cytokine secretion, the IFN-γ secretion increased, while the IL-10 secretion decreased in the sybiotic group. The result was agreed with previous study that the immunomodulating activity of L. rhamnosus GG was through upregulation of Th1 function.
Conclusion: The fermented plant fibers do have prebiotic effect which may stimulate the proliferation of fecal beneficial bacteria while inhibit the growth of fecal harmful bacteria. Therefore, we further added L. rhamnosus GG to these fermented plant fibers residues to develop a novel “synbiotic” which had better prebiotic effect than the fermented plant fibers alone.

目錄……………………………………………………………………1-6
縮寫對照表…………………………………………………………7-8
中文摘要…………………………………………………………9-10
英文摘要……………………………………………………………11-12
第一章前言及文獻回顧………………………………………13-23
第一部份 益生菌………………………………………………… 13
第二部份 益菌生…………………………………………………18
第三部份 共生菌…………………………………………………19
第四部份 植物纖維發酵物………………………………………21
第二章 實驗目的………………………………………………24-26
第三章 實驗材料與方法…………………………………………27-34
第一部份 實驗架構………………………………………………27
第二部份 發酵植物纖維錠樣品分析檢測……………………28-34
第一節 蔬果發酵後植物纖維益菌功能之評估……………28
第二節 蔬果發酵纖維錠中含菌量之測定…………………31
第三節 LGG乳酸桿菌蔬果發酵纖維錠安定性檢測………33
第三部份 人體試驗…………………………………………35-42
第一節 實驗設計……………………………………………35
第二節 糞便檢體腸道菌相之檢測…………………………38
第三節 血液一般生化檢驗…………………………………41
第四節 問卷調查……………………………………………42
第五節 統計方法……………………………………………42
第四部份 小鼠動物實驗……………………………………43-58
第一節 實驗設計……………………………………………43
第二節 腸道菌相之評估……………………………………44
第三節 免疫功能之研究……………………………………45
第四節 統計方法…………………………………………58
第四章結果與討論…………………………………………………59-74
第一部份蔬果發酵後植物纖維之分析檢測…………………59-62
第一節 蔬果發酵後植物纖維益菌功能之評估……………59
第二節 含菌量之測定………………………………………61
第三節 LGG蔬果發酵纖維錠中LGG益生菌安定性……62
第二部份 人體試驗…………………………………………63-68
第一節 受試者基本資料 & 問卷調查……………………63
第二節 血液一般生化檢驗…………………………………64
第三節受試者腸道細菌菌相的變化………………………65
第三部份 小鼠動物實驗……………………………………69-74
第一節 腸道菌相之研究……………………………………69
第二節 血液免疫功能評估…………………………………71
第五章結論…………………………………………………………75
表……………………………………………………………………76-80
圖……………………………………………………………………81-94
參考文獻………………………………………………………95-101
附件………………………………………………………………102-108

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