劉凱莉Liu, Kai-Li李珮毓Li, Pei-YuPei-YuLi營養學系碩士班2026-03-242029-08-2720242024-07-24https://handle.ncl.edu.tw/11296/8an9kwhttps://scholars.csmu.edu.tw/handle/123456789/7192潰瘍性結腸炎（Ulcerative colitis, UC）是發炎性腸道疾病的一種，在台灣UC盛行率逐年增加，目前臨床治療多以抗發炎藥物來治療及減少復發，然而UC的致病機轉尚不明確，可能是多重因子失衡所致，與黏膜屏障受損、發炎反應、氧化壓力相關。已知黃酮類化合物具有抗氧化及抗發炎功效，因此，本研究目的是探討白楊素及槲皮素在預防或改善dinitrobenzene sulfonic acid（DNBS）誘導結腸炎的保護潛力。五週齡雄性C57BL6/J小鼠適應一週後，依體重隨機分成控制組（C組）、DNBS組（D組）、白楊素（Chrysin, DC組）和槲皮素（Quercetin, DQ組）四組，DC組和DQ組分別灌食50 mg/kg體重的白楊素和槲皮素，C組和D組灌食相同體積的載體，管餵3週後由直腸輸注一次DNBS（200 mg/kg）誘發結腸炎，C組輸入等體積的50%乙醇，並連續三天記錄體重、收集糞便以評估疾病活動指數（Disease activity index, DAI），於誘發三天後犧牲動物，收集血液及結腸進行後續分析。結果顯示，灌食白楊素和槲皮素三週對小鼠體重沒有影響；結腸炎誘發後，在黏膜屏障受損的部分，相較於C組，D組顯著導致嚴重的疾病活動指數、結腸縮短、結腸黏膜與黏膜屏障受損，並影響細胞凋亡蛋白的表現（增加Bax及減少Bcl2）。然而，相較於D組，DC組及DQ組於DNBS誘發後第2天顯著改善DAI評分，且DC組改善DAI評分可延續至第3天。此外，DC組也顯著改善結腸縮短的情況、降低組織病理評分、並增加黏液蛋白MUC2、緊密連接蛋白（ZO-1、Occludin及E-Cadherin）的表現，並影響細胞凋亡蛋白的表現（Bax下降、Bcl2增加），DQ組則明顯增加MUC2、ZO-1、E-Cadherin和Bcl2的表現。在發炎反應的部分，相較於C組，D組明顯增加TLR4/MAPK/NF-κB）蛋白質及發炎物質（IL-6、IL-1β、TNF-α、IFN-γ、iNOS與COX2）mRNA的表現以及血漿IL-6含量；相較於D組，DC組皆可顯著改善上述發炎因子的表現，DQ組則改善TLR4、p-ERK、p-IκBα、IκBα和發炎物質的表現。在氧化壓力的部分，相較於C組，D組明顯降低結腸抗氧化酵素蛋白質（HO-1、GCLM、GR、NQO-1及SOD1）和mRNA（Catalase、SOD2、GPx1、GPx2及GCLC）表現，並增加紅血球丙二醛（Malondialdehyde, MDA）含量；相較於D組，DC組皆可顯著提高上述抗氧化酵素的表現，DQ組則是顯著提高結腸HO-1和GCLM蛋白質表現，以及Catalase、SOD2、GPx1、GPx2及GCLC的mRNA表現，此外DC組和DQ組皆顯著降低紅血球MDA含量。另外，D組也明顯增加內質網壓力路徑蛋白質（Bip、p-PERK、p-IRE1α、ATF4及CHOP）與CHOP之mRNA的表現，DC組可明顯改善上述蛋白質的表現，而DQ組僅降低p-PERK、ATF4及CHOP蛋白質與CHOP之mRNA表現。此外，D組血清指標也明顯高於C組，包括高靈敏度C反應蛋白（hsCRP）、AST、ALT、Creatinine和Uric acid，補充白楊素可顯著改善上述血清指標，而補充槲皮素可顯著降低ALT、Creatinine和Uric acid。綜合以上結果可知，補充白楊素和補充槲皮素可以透過調節結腸黏膜屏障功能、減少發炎反應和內質網壓力、以及增加抗氧化作用，同時改善血液發炎反應及氧化壓力，達到預防結腸炎病程的發展。Ulcerative colitis (UC) is an inflammatory bowel disease with increasing prevalence in Taiwan. Clinical treatments mainly use anti-inflammatory agents to reduce recurrence. The pathogenesis of UC is unclear but may involve mucosal barrier damage, inflammatory responses, and oxidative stress. Flavonoids have antioxidant and anti-inflammatory properties. Therefore, this study aims to investigate the protective potential of chrysin and quercetin in preventing or ameliorating dinitrobenzene sulfonic acid (DNBS)-induced colitis. Five-week-old male C57BL6/J mice were stabled for one week and then randomly divided into four groups by body weight: control group (C group), DNBS group (D group), chrysin group (DC group), and quercetin group (DQ group). DC and DQ groups were orally administered chrysin and quercetin at a dose of 50 mg/kg body weight, respectively. C and D groups received the same volume of vehicle. After three weeks of gavage, colitis was induced by rectal administration of DNBS (200 mg/kg), while C group received an equivalent volume of 50% ethanol. Body weight was recorded, and fecal samples were collected to assess disease activity index (DAI) daily for three days post-induction. Mice were sacrificed on the third day post-induction, and blood and colon were collected for analysis. Results showed that treatment with chrysin and quercetin did not affect body weight in mice. After colitis induction, compared to C group, D group exhibited significantly increased DAI scores, colon shortening, mucosal damage, and altered expression of apoptosis-related proteins (increased Bax and decreased Bcl2). However, compared to D group, DC and DQ groups significantly improved DAI scores by the second day after DNBS induction, with DC group maintaining improvement through the third day. Additionally, DC group significantly mitigated colon shortening, reduced histopathological scores, increased expression of MUC2, tight junction proteins (ZO-1, Occludin, and E-Cadherin), and altered apoptosis protein expression (decreased Bax, increased Bcl2). DQ group significantly increased MUC2, ZO-1, E-Cadherin, and Bcl2 expression. In terms of inflammatory response, DNBS significantly upregulated TLR4/MAPK/NF-κB pathway proteins, inflammatory mediators (IL-6, IL-1β, TNF-α, IFN-γ, iNOS, and COX2 mRNA expression), and plasma IL-6 levels. Compared to D group, DC group significantly ameliorated expression of these inflammatory factors, while DQ group improved TLR4, p-ERK, p-IκBα, IκBα and inflammatory mediators expression. Regarding oxidative stress, compared to C group, D group significantly decreased colonic anti-oxidant enzyme protein and mRNA expression, and increased red blood cell malondialdehyde (MDA) levels. Compared to D group, DC group significantly increased expression of antioxidant enzyme and decreased red blood cell MDA levels. DQ group significantly increased colonic HO-1 and GCLM protein expression, as well as mRNA expression of Catalase, SOD2, GPx1, GPx2, and GCLC. Furthermore, both DC and DQ groups significantly decreased red blood cell MDA levels. Moreover, D group significantly increased endoplasmic reticulum (ER) stress pathway proteins (Bip, p-PERK, p-IRE1α, ATF4, and CHOP) and CHOP mRNA expression, which was significantly improved in the DC group, whereas DQ group only decreased p-PERK, ATF4, and CHOP protein, and CHOP mRNA expression. Additionally, serum markers in D group were significantly higher than those in C group, including high-sensitivity C-reactive protein (hsCRP), AST, ALT, creatinine, and uric acid. Chrysin significantly improved these markers, while quercetin significantly decreased ALT, creatinine, and uric acid levels. To sum up, chrysin and quercetin may prevent the progression of colitis by regulating colonic mucosal barrier function, reducing inflammatory responses and ER stress, increasing antioxidant activity, and improving blood inflammatory response and oxidative stress.疾病活動指數DNBS槲皮素白楊素黏膜屏障Disease activity indexquercetinchrysinmucosal barrierthesis