臺灣博碩士論文加值系統

背景:
在科技的進步下和第二代高通量定序儀(NGS)的出現，使腸道菌群的研究有了大大的進展，在近期研究發現腸道微生物菌群與癌症、代謝疾病都有關聯。改變腸道微生物菌群的變因有非常多，其中主要的有飲食習慣以及生活作息，而運動以及熱量的攝取也是改變腸道微生物菌群的因素之一，因此本研究以老鼠為模式探討有氧運動、阻抗運動、高脂飲食和卡洛里限制下的微生物菌群之變化，此外腸道微生物菌群影響宿主健康的原因可能是腸道微生物的代謝產物，例如:短鏈脂肪酸等。因此本研究使用碳13標定追蹤碳源，探討宿主細胞受到腸道菌群代謝物之調控
實驗設計:
分別選用了營養介入的高脂飲食組和卡洛里限制組，運動模式介入的有氧運動組和阻抗運動組，分別挑選氧運動、阻抗運動、卡洛里限制組，使用NGS定序16s rRNA分析老鼠糞便之腸道菌群。接著各組分別透過13C-Glucose、13C-Serine的標定下，瞭解宿主和不同腸道微生物菌群運用葡萄糖或絲氨酸合成丁酸等短鏈脂肪酸。並在不同微生物菌群下，使用13C-Glucose將菌群體外培養，並追蹤不同模式下菌體的檸檬酸循環。
結果:
我們的結果發現在血液中有氧運動組丁酸和卡洛里限制組都有著師趨勢增加，而阻抗運動組的丁酸則不影響。糞便菌群分析後發現有氧運動組的丁酸生產菌Clostridium_sensu_stricto_1、Lachnospiraceae、Ruminococcaceae並沒有顯著增加。在體外培養中卻發現有氧運動組乙酸不改變，丁酸濃度增加，阻抗運動組乙酸丁酸都是下降，在高脂飲食組中乙酸增加丁酸減少，卡洛里限制組丁酸趨勢下降。因此從我們的實驗方法可以了解行為和營養介入下，腸道菌群對於不同碳源的使用與代謝產物和宿主的交互作用。從這兩個結果發現乙酸和丁酸在腸道菌群的改變後，不一定直接如同菌相預期丁酸菌增加血液丁酸就會增加，這其中可能與人體的代謝改變有關。

Background: Becase of science and technology improved and Next Generation Sequencing (NGS). In recent studies, it has been found that gut microbiota is associated with cancer and metabolic diseases. The gut microbiota may be changed by many factors. The main ones are eating habits, lifestyle, exercise and calorie intake are also one of the factors that change the gut microbiota. Therefore, we used mice as a model to explorec aerobic exercise, resistance exercise, high-fat diet and calorie restriction to changes in gut microbiota. In addition, the cause of gut microbiota affecting host health may be microbiota metabolites. For example, short-chain fatty acids (SCFA). Therefore, this study used isotope to track carbon sources and explored the regulation of host by gut microbiota metabolites.
Methods: The exercise mode had the aerobic exercise group and the resistance exercise group. The nutritional mode had high-fat diet group and the calorie restriction group. We were selected the aerobic exercise group, resistance exercise group and calorie restriction group to analyzed using NGS sequencing 16s rRNA. Then, each group was used by 13C-glucose and 13C-Serine to understand that the different gut microbiota synthesized SCFA such as butyrate by glucose or serine in the host. Under different gut microbiota, 13C-glucose was used to culture the bacteria in vitro, and the different modes have different citric acid cycle's carbone flow.
Result: Our results found that aerobic exercise group and calorie restriction group both improved the butyrate in plasma.But resistance exercise group hadn't changed of butyrate in plasma. After analysis of the fecal microbiota, the butyrate producing bacteria Clostridium_sensu_stricto_1, Lachnospiraceae, and Ruminococcaceae in the aerobic exercise group were not significantly increased. In vitro, we were found that the aerobic exercise group did not change acetate concentration, but butyrate concentration increased. The esistance exercise group decreased the acetate and butyrate. The igh-fat diet group increased acetate but decreased butyrate. The caloric restriction group decrease butyrate. Therefore, our experimental methods can understand the interaction of the gut microbiota used different carbon sources to interaction between metabolites and hosts, under exercise mode and nutritional mode. So we found that the changes of acetate and butyrate in the gut microbuita were not necessarily directly increased as the plasma, which may be related to the metabolic changes of the human body.

目錄
1. Part I. 運動模式和營養模式改變腸道微生物組成和影響宿主代謝 i
摘要 i
Abstract ii
目錄 iii
表目錄 iv
圖目錄 vi
第一章 前言 1
第二章 材料與方法 7
第三章 結果 12
第四章 討論與結論 20
Part1.table 29
Part 1.Figure 43
2. Part II. 短鏈脂肪酸對於肝癌和模擬正常肝細胞代謝影響 54
摘要 54
Abstract 55
第一章 前言 56
第二章 材料與方法 58
第三章 結果 61
第四章 討論與結論 70
Part2. Table 73
第五章 參考文獻 132

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