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研究生:劉宇碩
研究生(外文):LIU, YU-SHUO
論文名稱:補充魚油及雲芝胞外蛋白聚醣對短腸症候群大鼠之腸道損傷、發炎反應和腸菌相的影響
論文名稱(外文):The effects of fish oil and Trametes versicolor extracellular proteoglycan on intestinal damage, inflammatory response and dysbiosis in rats with short bowel syndrome
指導教授:羅慧珍
指導教授(外文):LO, HUI-CHEN
口試委員:徐泰浩李建興
口試委員(外文):HSU, TAI-HAOLEE, CHIEN-HSING
口試日期:2020-06-10
學位類別:碩士
校院名稱:輔仁大學
系所名稱:營養科學系碩士班
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2020
畢業學年度:108
語文別:中文
論文頁數:165
中文關鍵詞:短腸症候群魚油雲芝胞外蛋白聚糖腸道損傷發炎反應腸道菌
外文關鍵詞:short bowel syndromefish oilTV LH-1 ePGintestinal damageinflammatory responseintestinal microbiota
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短腸症候群(SBS)因大量腸切除造成腸道長度不足,導致吸收不良並伴隨腹瀉、脫水及腸蠕動受損等症狀發生,故SBS患者須以全靜脈營養(TPN)維持營養狀態。然而,SBS及長期使用TPN易造成腸黏膜萎縮、發炎反應及腸道菌改變。魚油已被證實具抗發炎、調節免疫與腸道菌的作用;臨床試驗證實重症患者給予靜脈魚油乳劑具調節其發炎與免疫反應、降低感然率及縮短住加護病房與住院時間等效益。在細胞與動物實驗證實雲芝胞外蛋白聚糖(TV LH-1 ePG)具免疫調節、降血糖和改善骨質密度的作用。因此本研究目的為探討口服補充魚油和TV ePG對SBS大鼠腸道損傷、發炎反應及腸道菌的影響(第一部分),並比較給予口服或靜脈補充魚油乳劑及合併口服TV ePG之效果(第二部分)。Sprague–Dawley (SD)大鼠經右頸靜脈插管和空-結腸吻合手術,術後立即給予TPN輸液7天。術後第一天,第一部分分別給予SBS大鼠單獨口服補充大豆油、魚油和TV ePG及合併給予大豆油或魚油和TV ePG;第二部分分別給予口服或靜脈補充魚油與口服TV ePG,及合併給予口服或靜脈補充魚油和TV ePG。術後第七天對SBS大鼠進行犧牲,採集血液和器官做後續實驗分析。實驗結果顯示,SBS大鼠有腹瀉、體重下降、器官腫大(脾、肝和肺)及血脂異常等問題,且血液IL-6和IL-8濃度及空腸IL-6含量的增加會造成全身性與腸道發炎;SBS大鼠的腸道菌數量及多樣性下降,且腸道菌群落改變(優勢菌種由Firmicutes轉換為Proteobacteria。給予口服或靜脈補充魚油或合併口服TV ePG可降低SBS大鼠的空腸IL-6和IL-8含量,但無法緩解全身性發炎與改善腸道菌紊亂;合併給予口服補充魚油和TV ePG會造成SBS大鼠腸道損傷。口服補充TV ePG可降低SBS大鼠的空腸和血漿IL-6和IL-8含量,且口服補充TV ePG可增加產生丁酸的腸道菌Lachnospiraceae和Ruminococcaceae之豐富度,此顯示TV ePG可能具有益生元的特性。綜上所述,大量腸切除後合併給予口服或靜脈補充魚油和TV ePG無法改善SBS的問題,且不建議合併給予口服魚油和TV ePG以避免造成腸道損傷,而單獨給予口服TV ePG具有調整SBS大鼠腸道菌相與緩減腸道與全身性發炎之功效。
Short bowel syndrome (SBS) is the symptoms with massive bowel resection following a deficiency of intestinal length that cause malabsorption and accompany with diarrhea, dehydration and impaired intestinal motility. SBS patients were total parental nutrition (TPN)-dependent to maintain nutritional status. However, SBS and long-term TPN is prone to cause intestinal injury, inflammatory response and dysbiosis. Fish oil had been evaluated abilities with anti-inflammation, immunomodulation and modulation of intestinal microbiota. Parenteral fish oil lipid emulsion had been used for critically ill patients in clinical trail, which associated with regulation of inflammation and immune responses, decline of infection rate and reduction in length of intensive care unit (ICU) and hospital stay. Trametes versicolor LH-1 extracellular proteoglycan (TV LH-1 ePG) had been evaluated abilities with immunomodulation, reduction of glycemic and improvement of bone density in cell line and animal studies. Therefore, this study aimed to investigate the effects of oral fish oil and TV ePG on intestinal injury, inflammatory response and dysbiosis in rats with SBS (part 1) and comparate the effects of parenteral infusion or ora of fish oil with TV ePG (part 2). Sprague–Dawley (SD) rat underwent right jugular vein catheterization and jejunocolostomy, and immediately administered TPN infusion after surgery until on postoperative day 7. On postoperative day 1, SBS rats were orally administered with soy bean oil, fish oil and TV ePG, and soy bean oil or fish oil with TV ePG in part 1 of the study; In part 2 of the study, SBS rats were orally or parenterally administered with fish oil and orally administered with TV ePG, and orally or parenterally with fish oil and TV ePG. All rats were sacrificed to harvest blood and organs in postoperative day 7. The results showed that SBS rats had diarrhea, loss body weight, hypertrophy of organs (spleen, liver and lung) and hyperlipidemia. SBS rats had systemic and intestinal inflammation which may cause by increasion of IL-6 and IL-8 concertation in blood and increasion of IL-6 in intestine. The results showed that reduced number and of intestinal microbiota in SBS rats, and alterated intestinal microbiota community which the dominant intestinal microbial phyla shift Firmicutes to Proteobacteria. SBS rats orally and parenterally administered with fish oil that reduced the SBS-increased intestinal IL-6 and IL-8, it could not alleviated systemic inflammation and dysbiosis in rats with SBS, and intestinal damage may cause by orally administered with fish oil. SBS rats orally administered with TV ePG which significantly decreased the SBS-increased intestinal and plasma IL-6 and IL-8 and increased the abundance of Lachnospiraceae and Ruminococcaceae which produce butyric acid. It was indicated that TV ePG may have the potential effects as prebiotic. In summary, there were not beneficial effects of oral or parenteral fish oil with TV ePG in rat with SBS, and most important of all is no recommance oral fish with TV ePG after massive intestinal resection which may cause intestinal damage. single supplement of TV ePG have the beneficial effects on regulation of intestinal microbiota and alleviation of intestinal and systemic inflammation in rat with SBS.
目錄
摘要 I
Abstract III
目錄 V
圖目錄 IX
表目錄 XII
謝誌 XIV
縮寫 XVI
第一章 前言 1
第二章 文獻探討 3
第一節 短腸症候群 3
一、 吸收不良與腸道適應性反應 4
二、 大量腸切除導致發炎反應 6
三、 大量腸切除導致腸道菌改變 8
第二節 短腸症候群之臨床照護 11
一、 全靜脈營養支持的效益 11
二、 全靜脈營養支持的副作用 12
三、 新型脂肪乳劑 15
第三節 輔助治療 16
四、 魚油 16
五、 雲芝蛋白聚糖 19
第三章 假說與目的 22
第四章 材料與方法 24
第一節 研究設計 24
六、 雲芝胞外蛋白聚糖萃取前製備 24
七、 動物適應期 24
八、 SBS大鼠手術模式 25
九、 動物實驗設計 27
第二節 分析項目 31
十、 體重、組織重量、TPN輸液量及熱量 31
十一、 全血球計數及血液生化指標 32
十二、 空腸組織型態及質量分析 32
十三、 血漿與空腸免疫球蛋白A含量 34
十四、 全身及局部發炎媒介物含量 34
十五、 血漿脂肪酸代謝衍生物含量 36
十六、 腸道內容物短鏈脂肪酸含量 36
十七、 腸道內容物腸道菌分析 37
第三節 統計方法 39
第五章 結果 41
第一部份 41
第四節 體重、熱量攝取與器官相對重量 41
第五節 全血球計數與血液生化值 42
第六節 血漿發炎媒介物質 43
第七節 空腸形態及蛋白質和DNA含量 44
第八節 血漿與空腸IgA濃度 44
第九節 空腸發炎媒介物質含量 45
第十節 腸道菌分析 45
十八、 腸道菌的複雜度分析與群落結構之主座標分析 45
十九、 腸道菌門階層的物種豐富度 46
二十、 腸道菌各階層的物種豐富度 47
第十一節 腸道內容物的SCFAs含量 48
第二部份 48
第十二節 熱量攝取、體重變化與器官相對重量 48
第十三節 全血球計數與血液生化值 49
第十四節 血漿發炎媒介物質 50
第十五節 空腸形態及蛋白質和DNA含量 51
第十六節 血漿與空腸IgA濃度 51
第十七節 空腸發炎媒介物質含量 51
第十八節 腸道菌分析 52
二十一、 腸道菌的複雜度分析與群落結構之主座標分析 52
二十二、 腸道菌門階層的物種豐富度 53
二十三、 腸道菌各階層的物種豐富度 54
第十九節 道內容物的SCFAs含量 55
第七章 討論 56
第一節 短腸症大鼠 56
第二節 口服補充魚油對短腸症之腸損傷、發炎反應與腸道菌變化的影響 60
第三節 靜脈補充魚油乳劑對短腸症模式之腸損傷、發炎反應與腸道菌的影響 65
第四節 口服TV ePG對短腸症模式之腸損傷、發炎反應與腸道菌的影響 67
第五節 合併給予魚油和TV ePG對短腸症模式之腸損傷、發炎反應與腸道菌的影響 69
第八章 結論 71
第九章 參考文獻 72
第十章 圖 81
第一部份 81
第二部份 99
第十一章 表 114
第一部份 114
第二部份 124
附錄 134


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