臺灣博碩士論文加值系統

膳食纖維可調節腸內菌叢生態並維持腸道功能；而培耶氏斑是食物由口攝取後第一個接觸的淋巴結。膳食纖維是否可以刺激培耶氏斑淋巴球增生進而調節免疫能力？本研究目的是探討餵食不同種類和比例膳食纖維飼料對於Wistar雄鼠培耶氏斑淋巴球增生與免疫反應。剛離乳Wistar雄鼠分別以5%纖維素、5%米糠、10%纖維素、10%米糠及對照組 (未含纖維質) 餵食八週即犧牲。培耶氏斑淋巴球收集後添加伴刀豆球蛋白 (concanavalin A) 和脂多醣 (lipopolysaccharide) 並進行細胞培養48小時後細胞液以WST-1試劑偵測細胞內粒腺體脫氫酶 (mitochondria dehydrogenase)之活性；細胞上清液則以酵素連結免疫分析來定量免疫球蛋白 (immunoglobulin)、細胞激素中介白素 (interleukin) 之IL-2、IL-6及腫瘤壞死因子 (tumor necrosis factor；TNF-α)。另外培耶氏斑淋巴球中B細胞收集後分為三組，對照組，一組中添加LPS和IL-4組，另一組則添加CD40和IL-7，隨即進行細胞培養48小時後萃取細胞中RNA以反轉錄聚合酶鏈合反應 (transcription polymerase chain reaction, RT-PCR) 來定量重組活化基2 (recombination activating gene2, RAG2) 之基因表現。結果顯示：細胞增生能力試驗中，所有實驗組皆高於對照組，5%米糠和纖維素組高於10%米糠和纖維素 (p<0.05)，但5%米糠與5%纖維素之間並無顯著差異。免疫球蛋白A、介白素和腫瘤壞死因子試驗中，所有實驗組皆高於對照組且5%米糠和纖維素高於10%米糠和纖維素 (p<0.05)。然而RAG2基因表現定量中則無產物表現。膳食纖維可提升腸道區域性淋巴球之增生能力和細胞激素濃度，接著細胞激素可誘導Ｂ細胞活化後分化為漿細胞且分泌大量免疫球蛋白A。結論為膳食纖維可調節淋巴細胞的成熟與分化；適量膳食纖維可能為抗原以誘發免疫機制的啟動。

Dietary fiber can modulate in the growth and distribution of fungus in intestinal tract function well. Peyer’s patches are the kind of lymph nodes that are the first contacted by food molecules after ingestion. May dietary fiber stimulate the proliferation of Peyer’s patches’ lymphocytes and regulate the immune response？ The purpose of this study is proliferation and immune response of Peyer’s patches’ lymphocytes in Wistar male rats treated with different types and proportion of dietary fiber. Wistar rats were divided in to 5 groups that were treated with 5% (w/w) cellulose, 5% rice bran, 10% cellulose, and 10% rice bran in the feeding respectively as well as control group without any fiber addition in the feeding. The Wistar rats were treated for 8 weeks from 3 weeks old to 11 weeks old. After sacrificing, the lymphocytes of Peyer’s patches (PPs) were collected and cultured with Concanavalin A (Con A) and Lipopolysaccharide (LPS) for 48 hours. The index of lymphocytes proliferation was the activity of mitochondria dehydrogenase detected by WST-1 [2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-
(2, 4-disulfophenyl)-2H-tetrazolium, monosodium salt]. The supernatant of cell culture was prepared for the level of immunoglobulin A (IgA), interleukin 2 (IL-2), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α) analyzed with enzyme-linked immunosorbent assay. Total RNA was extracted from B lymphocytes treated with interleukin 7 (IL-7) & CD40, and LPS & interleukin 4 (IL-4), respectively. The expression of recombination activating gene was then amplified with reverse transcription-polymerase chain reaction (RT-PCR). The proliferation activity in all fiber-treated groups was significantly higher than that in control group. The proliferation activity in 5% rice bran and cellulose-treated groups was significantly higher than that in 10% rice bran and cellulose treated groups (p<0.05). The proliferation activity of 5% rice bran not different from 5% cellulose. The level of IgA, IL-2, IL-6, and TNF-α in all fiber-treated groups was significantly higher than the control group. Similarly, IgA, IL-2, IL-6, and TNF-α levels in 5% rice bran and cellulose-treated groups were significantly (Statistically) higher (greater) than those in 10% rice bran and cellulose-treated groups. Dietary fiber may induce the secretion of cytokines, for example IL-2 and IL-6 from T cell than promote B cell to differentiate into plasma cells that could manufactory immunoglobulin. At the same time, lymphocytes in PPs proliferation may be induced by treatment of dietary fiber. In conclusion, dietary fiber may modulate the proliferation and immune response of PPs in intestinal tract. Also the appropriate content of dietary fiber was the important factor for the immunity modulated by dietary fiber.

表目錄
圖目錄
中文摘要......................................................I
英文摘要.....................................................II
第一章 前言..................................................1
第二章 文獻回顧..............................................3
第一節 膳食纖維(Dietary Fiber)之介紹.........................3
第二節 腸管上皮黏膜組織之介紹................................8
第三節 重組活化基因2(Recombination Activating Gene2, RAG2)蛋白
之簡介...............................................11
第四節 細胞激素對周邊淋巴球之影響作用.......................17
第五節 膳食纖維對B細胞及T細胞之影響.........................20
第三章 研究動機與目的.......................................25
第四章 材料與方法...........................................27
第五章 結果.................................................37
第六章 討論.................................................41
參考文獻.....................................................58
附錄一 ELISA 原始數據
IL-2....................................................64
IL-6...................................................65
TNF-α.................................................66
Ig A...................................................67
附錄二 基因序列及引子位置
大鼠之GAPDH基因.........................................68
大鼠之RAG2基因..........................................69
小鼠之RAG2基因..........................................70

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