植生性乳酸菌對腸內菌相分佈之影響暨植化素抑制結直腸癌SW480細胞增殖作用

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Title:	植生性乳酸菌對腸內菌相分佈之影響暨植化素抑制結直腸癌SW480細胞增殖作用 Effect of lactic acid bacteria from fermented mustard on intestinal microflora distribution and antiproliferative action of phytochemicals on colorectal adenocarcinoma SW480 cells
Authors:	蕭甄庭
Contributors:	保健營養系陳師瑩杜平悳
Keywords:	植生性乳酸菌腸道機能 5-FU 虎杖海巴戟天甜橙結直腸癌協同作用 Lactic acid bacteria Fermented mustard Intestinal function 5-FU Polygonum cuspidatum Morinda citrifolia Citrus sinensis Colorectal carcinoma cell Synergism effect
Date:	2016
Issue Date:	2016-12-21 15:34:58 (UTC+8)
Abstract:	本研究探討植生性乳酸菌 B0091 於豆粉培養基中發酵後產物，對於改善腸胃道菌叢分布之影響。10週齡雄性 Wistar 系試驗鼠35隻，經一週環境適應後，隨機分成5組，每組7隻，依序為控制組 (Control)，正控制組(Casei, 管餵L. Casei, 1× 1011 CFU/kg bw)，以及實驗組 3 組，分別管餵三種不同劑量之植生性乳酸菌 B0091，分別為 B0091_L 組 (低劑量組：4× 109 CFU/kg bw)、B0091_M組 (中劑量組：2×1010 CFU/kg bw)、B0091_H組 (高劑量組：1× 1011 CFU/kg bw)。結果顯示每公斤體重試驗鼠管餵高劑量組植生性乳酸菌 (1× 1011 CFU /kg bw ) 於5週後具有顯著改善Bifidobacteria / Cl. perfringens菌叢分布的效果，再以Bifidobacteria 和Cl. perfringens的DNA分析檢測，結果亦顯示高劑量組 (B0091_H) 之Bifidobacteria / Cl. perfringens比值顯著高於控制組與正控制組 (Casei)；針對試驗鼠空腸進行切片觀察絨毛長度與腺窩深度及其結構變化，結果顯示B0091_H 絨毛長度與腺窩深度的比值顯著優於控制組與Casei組，亦高於B0091_L組與B0091_M組。綜合以上結果顯示管餵高劑量 (1× 1011 CFU/kg bw) 植生性乳酸菌B0091，能顯著改善腸胃道菌叢分布及改善空腸生理結構。而結直腸癌在台灣癌症相關死亡原因為第三名。為?提高癌症預後，?床治?皆使用化?；但化?對正常組織所產生的毒性，已嚴重阻礙癌症的治?效果。故在化?過程中給予適當的輔助?法，減緩病人的?適，?僅可提高治癒機會，也減少化?藥物的毒性。5-FU 一直是治?結直腸癌的主要化?藥物，但治?效?一直維持在 20-35%，其中位?存活??超過 1 ?。因此，尋找具有高保健功效的抗癌成分，並與化?藥物組合運用，用以提高?好的治?效果，是治?結直腸癌的新興發展方向。研究顯示虎杖根部乙醇萃取物分劃物 (PcE(H)EA)、海巴戟天葉乙醇萃取物 (LES)、橙皮水萃物(WESP) 含有多樣化與高含?的多酚?物質，具有抗發炎和抗氧化生物活性，值得進一步純化與評估防癌功效。本研究主要探討 PcE(H)EA 和 LES 及 WESP 其活性成分對結腸癌細胞 SW480 體外試驗的抑制效果。依據抑制細胞增生的效率、半抑制率濃度 (IC50) 及聯合效應 (CI) 等參數，驗證抑制結直腸癌細胞增生的效果，最佳順序為：Resveratrol 聯合 Emodin 在濃度 (w/v) 比例為 1：4 的組合>Emodin>PcE(H)EA>Hesperetin>Hesperidin>WESP>Resveratrol>F1>Rutin>LES，此結果將有助提供可?的輔助治?策?，兼具避免化?藥物被腫瘤細胞快速發展成抗藥性的現象。 The purpose of this study was to evaluate the probiotic properties of lactic acid bacteria (B0091) strain isolated from fermented mustard in vivo. Male Wistar rats (10-week-old) were randomly divided into three experiments groups (7 rats/group) that were high dose (1× 1011 CFU/kg bw), median dose (2× 1010 CFU/kg bw) and low dose (4× 109 CFU/kg bw) of B0091 were gavage for 5 consecutive weeks. Furthermore, the positive control was orally-administered L.casei (1× 1011 CFU/kg bw), the control was treatment with the normal saline instead of sample administration. All animals were acclimatized for 1 week prior to use. The body weight and food intake were recorded daily. Fecal and fecal of cecum samples were collected to determine number of beneficial bacteria bifidobacterium and harmful bacteria Clostridium perfringens by classical plate count and by PCR amplification. Strain B0091 had a significant improve intestinal microflora distribution compared to the control. In addition, the villus height (vh), crypt depth (cd) and vh/cd ratio of rats’ jejunum were determined. The vh and vh/cd ratio were increased, which may suggest strain B0091 modulated intestinal mucin composition. Collectively, these results obtained from in vivo experiments positive effect of strain B0091 on antibacterial activity and thereby leading to improvement of intestinal health. Colorectal carcinoma is the third leading cause of cancer-related death in Taiwan. In order to improve the prognosis, chemotherapy is often used in a variety of clinical situations. The toxicity of these chemotherapeutic agents to normal tissues has been one of the major obstacles to successful cancer chemotherapy. Therefore, combined treatments with adjuvant therapy are often used not only to enhance the treatment effect, but also to reduce the toxicity of these drugs. 5-fluorouracil (5-FU) has been the major chemotherapeutic agent for treating colorectal carcinoma; however, response rates have been around 20–35% with median overall survival no more than 1 year. So finding anti-cancer compounds with high nutraceutic effect which can be used in combination with existing drugs may provide an important way forward in the treatment of colorectal carcinoma. Based on our researches, fractionation in ethanolic extracts from Polygonum cuspidatum root (PcE(H)EA) , Supernatant from the 95% ethanolic extracts of Morinda citrifolia leafexhibit (LES) , and water extracts of sweet orange (Citrus sinensis) peel (WESP) exhibit various and higher contents in polyphenols, and reveal that these compounds play a part in antioxidative and anti-inflammatory activity in vitro. Hence, PcE(H)EA, LES, WESP and its active components are worth further research in purification, and deeply investigation for anti-cancer effects. This study aim to investigate the inhibitive effects of active components from PcE(H)EA, LES, WESP combined with 5-FU on colorectal carcinoma (SW480) cells in vitro. The inhibition effects of cell proliferation of test sample and 5-FU, either in combination or on their own, are will be observed. According to the inhibition rate of cell viability, half maximal inhibitory concentration and combination effect, results in the order of efficacy showed that resveratrol plus emodin at concentration ratio of 1：4 (w/w)>Emodin>PcE(H)EA>Hesperetin>Hesperidin>WESP>Resveratrol>F1>Rutin>LES with 5-FU in SW480 cells. This is a very practicable strategy of adjuvant therapy—they might avoid the rapid development of resistance that occurs to drugs that target cancer cells.
Relation:	校外公開：2021-09-09,校內公開：2021-09-08，學年度：104，118頁
Appears in Collections:	[Dept. of Health and Nutrition (including master's program)] Dissertations and Theses

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