金針菇免疫調節蛋白 FIP-fve 生物可及性之研究

Bioaccessibility Study for Fungal Immunomodulatory Protein FIP-fve

劉育靈(Yu-Ling Liu)

指導教授 ： 許輔

國立臺灣大學/生物資源暨農學院/園藝暨景觀學系/碩士(2013年)

https://doi.org/10.6342/NTU.2013.10511

摘要

FIP-fve 為純化自金針菇之免疫調節蛋白，研究指出 FIP-fve 可刺激人類周邊血淋巴細胞增生及分泌 IFN-γ，並使免疫反應趨向於 TH1 反應；體內試驗亦發現，口服 FIP-fve 可降低小鼠食物過敏的症狀、延長罹癌小鼠之壽命及抑制腫瘤大小。生物可及性 (bioaccessibility) 是指活性成分自食物基質釋出後，可通過腸胃道的部分，用來評估活性成分的可利用性，其大多採取體外模擬腸胃道消化的試驗來證明。本實驗欲探討金針菇免疫調節蛋白 FIP-fve 經模擬腸胃道消化後之穩定性，並評估其是否具有活化腸道免疫細胞之功能。第一部分，在模擬胃、腸兩階段各別消化中，以含有胃蛋白酶之模擬胃消化液處理，結果顯示 FIP-fve 並不會有所降解，而在含有胰酶之模擬腸道消化液中，FIP-fve 雖少部分被消化並產生兩個片段，但大部分並未被消化。使用模擬連續式腸胃道消化 FIP-fve，其經模擬胃消化兩小時後，再進行模擬腸消化兩小時，透過高效能液相層析儀分析結果顯示 FIP-fve 並無降解。第二部分，實驗發現 FIP-fve 可以刺激腸繫膜淋巴結 (mesenteric lymph node, MLN) 及培耶氏斑 (Peyer’s patches, PP) 細胞產生 IFN-γ，此外，亦發現消化後的 FIP-fve 仍具有調節腸道免疫之活性，可刺激培耶氏斑細胞分泌 IFN-γ，因此推論 FIP-fve 之口服穩定性佳，模擬腸胃道消化並不會使 FIP-fve 降解，可通過消化道到達小腸，具良好之生物可及性。且經消化之 FIP-fve尚有調節腸道免疫之功能，可能進而影響全身性的免疫反應，達到治療疾病的效果。

關鍵字

FIP-fve ； 生物可及性 ； 體外模擬腸胃道消化 ； 腸繫膜淋巴結 ； 培耶氏斑

並列摘要

FIP-fve is an immunomodulatory protein which is purified from Flammulina velutipes. It has been investigated that FIP-fve has the abilities to stimulate the proliferation and the production of cytokine IFN-γ of human peripheral blood mononuclear cells (hPBMC) and skew the T cells toward TH1 immune response. Besides, in vivo studies indicated that the effects of orally administrated FIP-fve in decreasing food-allergic reaction in mice, inhibiting the progression of tumor size and prolonging the life of mice bearing liver cancer. Bioaccessibility stands for the availabilities of bioactive compounds released from food matrix after passing through gastrointestinal tract which is determined by mimicking digestive system in vitro. In this study, we would like to investigate the stability of immunomodulatory protein FIP-fve after passing through gastrointestinal digestion and determine whether its digested product could further activate the cells of gut-associated lymphoid tissue (GALT).
Results showed that during mimicking each stage of digestion in stomach and intestine, FIP-fve was not degraded under the presence of digestive enzyme which contained pepsin. Moreover, after incubated in simulated intestinal fluid (SIF) which contained pancreatin, FIP-fve was slightly digested into fragments but not digested completely. In addition, the mimicked continuous gastrointestinal digestion of FIP-fve was performed for 2 hours. In this case, results determined via HPLC showed that FIP-fve was not degraded during the mimicked gastrointestinal digestion. Furthermore, digested FIP-fve could trigger the activation of mesenteric lymph node (MLN) and Peyer’s patch (PP) cells to generate IFN-γ. Results reveals the effects that the modulation of immune responses on GALT was achieved by stimulating IFN-γ production from Peyer’s patch.
Briefly, we could conclude that orally intake of FIP-fve is considerably stable to reach intestine against gastrointestinal digestion. In addition, the potential of immunomodulatory effects of FIP-fve may further affect systemic immune responses and has the possibility to be utilized on therapy studies.

並列關鍵字

FIP-fve ； bioaccessibility ； in vitro gastrointestinal digestion ； mesenteric lymph nodes ； Peyer’s patch

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