臺灣博碩士論文加值系統

非酒精性脂肪肝 (nonalcoholic fatty liver disease, NAFLD) 為一種與肥胖相關的慢性、進行性疾病。甘藷富含多種活性物質，已被發現具抗發炎、抗氧化和益生元活性之潛力，而透過發酵能夠有效增加其中功效性物質含量。艾克曼嗜黏蛋白菌(Akkermansia muciniphila)，為腸道最豐富單一物種，許多證據顯示其與肝臟發炎及肥胖等疾病具相關性，於疾病發生的情況下A. muciniphila的豐度大幅下降，並導致腸道屏障功能減弱。為了解結合發酵甘藷萃取物與A. muciniphila對於肥胖是否具加成作用，本研究篩選台灣甘藷品種與台灣人源分離之巴氏殺菌A. muciniphila菌株(AKK19011)，評估其組合物對於高脂飼料誘導肥胖小鼠之影響。實驗結果發現，相對於台農57號黃色甘藷及台農66號紅色甘藷，台農73號紫色甘藷具有最佳的益生元潛力，而透過乳酸菌株OFM-73及OFM-143混合菌株發酵紫甘藷，可以有效提升功效性物質如多酚、類黃酮含量，並降低還原糖含量，其中以吲哚乳酸、雪松素、異鼠李素及莨菪素提升最為顯著。此外，於肥胖小鼠實驗模型中可發現，發酵紫甘藷乙醇萃取物(fermented purple sweet potato, FSPE)可顯著降低由高脂飲食誘導之體增重及脂肪組織重量增加現象，並且能夠有效降低血清及肝臟三酸甘油脂含量，使其肝臟脂滴大小及數量顯著抑制，且FSPE對於白色脂肪組織細胞面積增生亦具有顯著的抑制效果。此外，FSPE及FSPE + AKK19011組可以顯著提升糞便中短鏈脂肪酸丙酸及丁酸含量。而FSPE可調控肝臟中與肥胖及發炎反應相關之PPARα、PPARγ、TNF-α及IL-10基因表現；FSPE + AKK19011組能夠有效將脂肪組織中，因高脂肪飲食改變之基因PPARγ、TNF-α、IL-10、PGC-1α及SREBP-1C調控至接近ND組之表現，並且發現經由FSPE及A. muciniphila誘導後，脂質代謝物如三酸甘油脂、磷脂質及磷脂酸等，容易導致肥胖及肝損傷的代謝產物含量顯著降低，綜合以上結果，FSPE能有效降低由高脂飲食所引起的肥胖及脂肪增生現象並減緩NAFLD之發生，表明FSPE具有作為治療由高脂飲食引起的NAFLD替代用藥之應用潛力。

Nonalcoholic fatty liver disease (NAFLD) is a chronic and progressive condition closely linked to obesity. Sweet potatoes are rich in bioactive compounds with anti-inflammatory, antioxidant, and prebiotic properties, and their fermentation can enhance the concentration of these functional substances. Akkermansia muciniphila, the predominant single species in the gut microbiota, has been implicated in liver inflammation, obesity, and other related disorders. This study aims to investigate the effects of fermented sweet potato extract and pasteurized A. muciniphila on obesity. The research involved screening different varieties of Taiwanese sweet potatoes and a human-derived strain of A. muciniphila (AKK19011) to assess their impact on high-fat diet-induced obesity in mice. The purple sweet potato showed the highest prebiotic potential compared to the yellow and red sweet potatoes. Fermentation of the purple sweet potato using a combination of lactic acid bacteria strains (OFM-73 and OFM-143) increased functional compounds and reduced reducing sugars. Notably, compounds such as indolelactic acid, cedrelopsin, isorhamnetin, and scopoletin were particularly prominent. In the obesity model, the administration of fermented purple sweet potato ethanol extract (FSPE) resulted in a marked reduction in body weight gain and adipose tissue accumulation induced by a high-fat diet. Additionally, FSPE effectively lowered serum and hepatic triglyceride levels, significantly inhibited the size and number of hepatic lipid droplets, and demonstrated a notable suppressive effect on the hyperplasia of white adipose tissue cells. Furthermore, both FSPE and the combination of FSPE with AKK19011 significantly elevated the levels of short-chain fatty acids, specifically propionic acid and butyric acid, in fecal samples. FSPE modulated the expression of genes associated with obesity and inflammation in the liver, including PPARα, PPARγ, TNF-α, and IL-10. The FSPE + AKK19011 group exhibited a regulatory effect on the expression of PPARγ, TNF-α, IL-10, PGC-1α, and SREBP-1C in adipose tissue, bringing their expression levels closer to those observed in the control group. Following the induction of A. muciniphila, there was a significant reduction in lipid metabolites such as triglycerides, phospholipids, and phosphatidic acids, which are associated with obesity and liver damage. These results suggest that FSPE can effectively mitigate obesity and adipose tissue hyperplasia induced by a high-fat diet and may slow the progression of NAFLD, indicating its potential as an alternative therapeutic agent for treating NAFLD associated with high-fat dietary intake.

壹、 前言 1
貳、 文獻整理 3
一、非酒精性脂肪肝(non-alcoholic fatty liver disease, NAFLD) 3
(一) 肥胖 3
1. 肥胖引起的腸道菌群失調 4
(二) 非酒精性脂肪肝之腸道微生物群 5
二、NAFLD的治療藥物及方法 6
(一) 運動控制 6
(二) 飲食改善 7
(三) 腸道微生物控制 7
三、甘藷〔Ipomoea batatas (L.) Lam〕 9
(一) 甘藷熱水萃取物 10
(二) 甘藷乙醇萃取物 10
(三) 發酵甘藷 11
(四) 抗氧化活性 13
(五) 抗發炎活性 14
(六) 抗肥胖活性 14
(七) 益生元活性 15
四、艾克曼嗜黏蛋白菌(Akkermansia muciniphila) 16
(一) 安全性及致病性 16
(二) 巴氏殺菌艾克曼嗜黏蛋白菌 17
參、 實驗設計 18
肆、 材料方法 19
一、 實驗原料 19
二、 實驗溶液配置 20
三、 實驗方法 22
(一) 樣品製備 22
(二) 甘藷品種之篩選 22
(三) 甘藷樣品之熱水萃取(Sweet potato hot water extract, SPE) 24
(四) 菌種篩選及甘藷樣品之發酵 25
(五) 發酵甘藷樣品之乙醇萃取(Fermented sweet potato ethanol extract, FSPE) 25
(六) 功效性物質分析 26
1. 總酚含量 26
2. 總類黃酮含量 26
3. 總花青素含量 26
4. pH 值 27
5. 還原糖含量測定 27
6. 發酵前後物質變化測定 27
(七) 動物實驗 28
1. 動物來源 28
2. 動物實驗流程圖 29
3. 紫甘藷熱水萃取物管餵劑量試驗 32
4. 小鼠糞便益生菌數測試 32
5. 甘藷熱水萃取物及艾克曼嗜黏蛋白菌動物實驗設計 32
6. 甘藷乙醇萃取物及艾克曼嗜黏蛋白菌動物實驗設計 33
7. 生理指標分析 34
8. 即時定量聚合酶連鎖反應測試(Real-time Quantitative Polymerase Chain Reaction, qPCR) 37
9. 糞便短鏈脂肪酸分析(Fecal short chain fatty acid analysis) 40
10. 脂質體學分析 41
(八) 統計分析 41
伍、 結果 42
一、 具有益生元潛力之甘藷篩選及其功效物質分析 42
二、 紫甘藷熱水萃取物對於C57BL/6小鼠之影響 45
三、 甘藷熱水萃取物及艾克曼嗜黏蛋白菌對於肥胖小鼠之影響 48
四、 發酵甘藷之功效性物質分析 50
五、 甘藷乙醇萃取物及巴氏殺菌艾克曼嗜黏蛋白菌對於高脂飲食誘導C57BL/6小鼠非酒精性脂肪肝之影響 56
六、 甘藷乙醇萃取物及巴氏殺菌艾克曼嗜黏蛋白菌對於高脂飲食誘導C57BL/6小鼠mRNA基因表現之影響 66
七、 甘藷乙醇萃取物及巴氏滅菌艾克曼嗜黏蛋白菌對於高脂飲食誘導C57BL/6小鼠代謝體學之影響 70
陸、 討論 75
柒、 結論 84
捌、 參考文獻 85
玖、 附錄 109

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