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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 潘敏雄(Min-Hsiung Pan) | |
| dc.contributor.author | Tai-Ling Chueh | en |
| dc.contributor.author | 闕岱伶 | zh_TW |
| dc.date.accessioned | 2023-03-19T21:30:43Z | - |
| dc.date.copyright | 2022-10-19 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-09-23 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/84072 | - |
| dc.description.abstract | 發炎性腸道疾病已被視為一項全球性的疾病，主要受到工業化國家生活方式西化及環境因子的影響。腸道屏障損傷是造成腸炎持續惡化的主因，而腸道屏障主要受到上皮緊密結合蛋白 (tight junction, TJ) 的支持。普遍而言，TJ 的損傷早於疾病進程，而腸道屏障功能喪失也受到促發炎細胞激素分泌及腸道菌相紊亂的影響，因此維持腸道屏障的完整性被視為腸炎的治療標的。雖然目前已有許多藥物用於治療腸炎，然而長期使用卻會引發諸多副作用，因此許多植化素已被作為腸炎的疾病預防策略。?類化合物 (stilbenoids) 是廣泛存在於葡萄、百香果、花生等植物中的酚類物質，已被證實具有抗氧化、抗發炎、抗癌等生物活性，其中又以白皮杉醇 (piceatannol. PIC) 及三羥基紫檀? (3′-hydroxypterostilbene, HPSB) 被認為具有相對良好的生物活性，然而兩者改善腸道屏障受損的相關機制仍有待釐清。因此本實驗旨在探討 PIC 及 HPSB 是否具有保護腸道屏障以緩解腸炎的能力，首先利用 TNF-α 誘導的細胞模型模擬腸道發炎環境，評估 PIC 及 HPSB 抗發炎及保護腸道屏障之潛力，再藉由 dextran sulfate sodium (DSS) 誘導的腸炎模式評估兩者調控分子機制、腸道菌相以改善腸炎的效果。細胞實驗結果顯示，PIC 與 HPSB 皆顯著下調 p-p65/p65 表現量，並且降低腸道上皮細胞單層的通透性。而在 DSS 誘導之 ICR 小鼠，則發現 PIC 與 HPSB 皆能顯著抑制 TNF-α/NF-κB/MLC 路徑及降低 NLRP3 發炎小體活化緩解發炎反應，但僅有 PIC 顯著調節 TJ 組成，推測可能與 PIC 更可顯著抑制腸道上皮細胞凋亡蛋白 bax/bcl-2 及 caspase-3 活化有關。從腸道菌相分析結果則觀察到 PIC 與 HPSB 皆能調節腸道菌相，提高 Akkermansiaceae、Lactobacillus intestinalis 等益菌，並下調 Spiroplasmataceae、Acholeplasmataceae 等害菌。由 LEfSe 分析則顯示介入 PIC 的組別，以丁酸鹽產生菌作為標識菌種，由此推測 PIC 尚具有增加短鏈脂肪酸合成之潛力，而可提供上皮細胞能量來源，以維持腸道屏障之構造。綜上所述，PIC 顯著降低發炎反應、抑制細胞凋亡，並調節腸道菌相，而相較 HPSB 更能夠維持腸道屏障的完整性，在未來具有作為腸炎化學預防製劑之潛力。 | zh_TW |
| dc.description.abstract | Inflammatory bowel disease (IBD) has emerged as a global disease, which is attributed to westernized lifestyle and other environmental factors in industrialized countries. Research has indicated that the disruption of gut barrier plays a crucial role in the development of colitis, and it is mainly supported by epithelial tight junctions (TJs). Commonly, TJs are impaired substantially precede the development of the disease. Besides, gut barrier dysfunction also affected by pro-inflammatory cytokines production and gut microbiota dysbiosis. Thus, maintaining gut barrier integrity is considered as the therapeutic target in colitis. Many synthetic drugs are currently in use to treat IBD; however, long-term usage of these drugs results in many complications. Hence, lots of phytochemical compounds have been proven as preventive strategy for IBD treatment. Stilbenoids are a group of phenolic compounds present in various plants, such as grape, passionfruit, and peanuts, which exert diverse bioactivities ranging from anti-oxidant, anti-inflammation, and cancer prevention. Among them, piceatannol (PIC) and 3′-hydroxypterostilbene (HPSB) are considered to have relatively good bioactivities. However, the mechanisms of PIC and HPSB on ameliorating gut barrier dysfunction haven’t been fully investigated. Therefore, we aimed to evaluate the ability of PIC and HPSB on gut barrier protection to reduce the severity of colitis. Firstly, we used TNF-α induced cell model to examine the anti-inflammatory and gut protection effect of PIC and HPSB, then used a DSS-induced colitis mice model to clarify the effect of both compounds on underlying mechanisms and microbiota modulation. Results obtained from in vitro experiments showed that, both of PIC and HPSB downregulated p-p65/p65 expression and decreased the permeability of intestinal epithelial monolayer. In contrast, in DSS-induced colitis ICR mice, both of PIC and HPSB could attenuate inflammation through inhibiting TNF-α/NF-κB/MLC pathway and reducing NLRP3 inflammasome activation. However, PIC was comparably effective in TJs modulation. The results may attribute to the effect of PIC on reducing cell apoptosis associated protein expression, including bax/bcl-2 and caspase-3 activation. Furthermore, the result of microbiota analysis showed that both of PIC and HPSB could increase the representative probiotics, including Akkermansiaceae and Lactobacillus intestinalis. On the other hand, supplementation of PIC and HPSB exhibited inhibitory effects on the several bacteria species (at family level) with undesired effect, including Spiroplasmataceae and Acholeplasmataceae. Based on LEfSe analysis, butyrate-producing bacteria was regarded as a biomarker in PIC group, so we speculated that PIC has the capability to increase short chain fatty acid production and can be used as an energy source for epithelial cell. In conclusion, the results demonstrated that PIC reduced inflammation, inhibited cell apoptosis, and regulated microbiota composition. Consequently, PIC is more effective on maintaining gut barrier integrity, and it will be a promising ingredient applied to the development of functional food for colitis prevention. | en |
| dc.description.provenance | Made available in DSpace on 2023-03-19T21:30:43Z (GMT). No. of bitstreams: 1 U0001-2109202201174400.pdf: 8890787 bytes, checksum: deece5f5932610c0dd2c53c49fccc397 (MD5) Previous issue date: 2022 | en |
| dc.description.tableofcontents | 口試委員會審定書 I 謝誌 II 摘要 IV Abstract V 目錄 VII 附圖索引 XI 附表索引 XII 圖目錄 XIII 縮寫表 XIV 第一章、 文獻回顧 1 第一節、 發炎性腸道疾病 1 (一)、 定義 1 (二)、 流行病學概況 2 (三)、 致病因素 5 (四)、 腸道發炎反應 6 (五)、 腸道屏障 8 (六)、 腸道屏障受損相關機制 11 (七)、 發炎小體活化對於發炎性腸道疾病之影響 12 (八)、 腸道上皮細胞凋亡對於發炎性腸道疾病之影響 14 (九)、 腸道菌相及短鏈脂肪酸與發炎性腸道疾病之關聯 16 (十)、 治療方式 18 第二節、 誘導結腸炎之動物模式 20 (一)、DSS 之特性與作用機制 20 (二)、DSS 之使用方式 21 第三節、 ?類化合物 (Stilbenoids) 22 (一)、 簡介 22 (二)、 白皮杉醇 (Piceatannol) 23 (三)、 三羥基紫檀? (3′-hydroxypterostilbene) 24 第二章、 實驗目的與架構 25 第一節、 實驗目的 25 第二節、 實驗架構 25 (一)、 細胞實驗 25 (二)、 動物實驗 26 第三章、 材料與方法 28 第一節、 實驗材料 28 (一)、 樣品及誘導劑 28 (二)、 藥品試劑 28 (三)、 分析套組 29 (四)、 實驗耗材 29 (五)、 儀器設備 29 (六)、 抗體 31 第二節、 細胞實驗 (in vitro) 方法 32 (一)、 細胞株 32 (二)、 培養液及試藥製備 32 (三)、 細胞培養 33 (四)、 樣品製備 35 (五)、 細胞存活率試驗 (MTT assay) 36 (六)、 細胞蛋白質萃取 37 (七)、 跨上皮電阻試驗 (Transepithelial electrical resistance, TEER) 38 (八)、 細胞旁通透性測試 39 第三節、 動物實驗 (in vivo) 方法 40 (一)、 動物品系與飼養環境 40 (二)、 動物實驗組別設計 40 (三)、 疾病活動指數 (Disease activity index, DAI) 41 (四)、 動物犧牲 43 (五)、 血液生化數值分析 43 (六)、 細胞激素測定 43 (七)、 腸道通透性試驗 46 (八)、 組織切片 46 (九)、 蘇木精-伊紅染色 (hematoxylin and eosin stain, H&E stain) 48 (十)、 免疫螢光染色 (Immunofluorescence stain, IF stain) 50 (十一)、 組織均質及蛋白質萃取 53 (十二)、 蛋白質定量 54 (十三)、 西方墨點法 54 (十四)、 微生物體全長 16S 定序分析 58 (十五)、 短鏈脂肪酸含量分析 59 第四節、 統計分析 61 第四章、 結果與討論 62 第一節、 細胞實驗 62 (一)、 ?類化合物與 TNF-α 對 NCM460 細胞毒性分析 62 (二)、 ?類化合物抑制發炎反應相關蛋白表現量之比較 64 (三)、 PIC 與 HPSB 對 TNF-α 誘導 Caco-2 腸道上皮細胞通透性 之影響 67 第二節、 動物實驗 70 (一)、 PIC 與 HPSB對 DSS 誘導之 ICR 小鼠體重、攝食量及飲水 量之影響 70 (二)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠結腸炎疾病活動指數 及受損程度之影響 72 (三)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠臟器重量之影響 77 (四)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠血清生化值之影響 80 (五)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠腸道通透性之影響 82 (六)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠腸道發炎之影響 86 (七)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠腸道上皮細胞凋亡 之影響 91 (八)、 PIC 與 HPSB 對DSS誘導之 ICR小鼠腸道菌相組成之影響 94 (九)、 PIC 與 HPSB 對 DSS 誘導之 ICR 小鼠結腸糞便短鏈脂肪酸 含量之影響 102 第五章、 結論 104 參考文獻 106 附錄 129 | |
| dc.language.iso | zh-TW | |
| dc.title | 白皮杉醇與3′-羥基紫檀?維持腸黏膜屏障完整性緩解發炎性腸道疾病之功效 | zh_TW |
| dc.title | Effects of piceatannol and 3′-hydroxypterostilbene on alleviating inflammatory bowel disease by maintaining intestinal epithelial integrity | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 110-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 何元順(Yuan-Soon Ho),黃步敏(Bu-Miin Huang),王應然(Yin-Jan Wang),郭靜娟(Ching-Chuan Kuo) | |
| dc.subject.keyword | 結腸炎,腸道屏障,緊密結合,NLRP3 發炎小體,白皮杉醇,三羥基紫檀?, | zh_TW |
| dc.subject.keyword | colitis,gut barrier,tight junctions,NLRP3 inflammasome,piceatannol,3′-hydroxypterostilbene, | en |
| dc.relation.page | 129 | |
| dc.identifier.doi | 10.6342/NTU202203694 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2022-09-25 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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