Epigallocatechin gallate-quercetin-rutin複方對ICR小鼠之抗疲勞作用與調控腸道菌相之研究

Effect of epigallocatechin gallate-quercetin-rutin formula on anti-fatigue and regulation of gut microbiota in ICR mice

丁頤瑄(Yi-Hsuan Ting)

指導教授 ： 徐慶琳

中山醫學大學/健康管理學院/營養學系/碩士(2020年)

https://doi.org/10.6834/csmu202000282

摘要

疲勞之定義為充滿疲倦感、缺乏活動力和無法維持自願性之身體活動，並且會對運動強度、工作表現、家庭生活和社會關係造成負面影響。目前有許多預防及改善疲勞之方法，其中之一為攝取機能性成分補充劑。過去文獻指出，腸道菌相組成與疲勞之發生有所相關，良好之腸道菌相組成具有提升運動表現之作用。天然多酚已知具有抗氧化、抗發炎、抗肥胖和抗癌等作用，但天然多酚複方在抗疲勞作用與腸道菌相之相關研究仍缺乏。因此，本研究將組合已知對於疲勞具有改善作用之表沒食子兒茶素沒食子酸酯 (epigallocatechin gallate, EGCG)、槲皮素 (quercetin) 與芸香苷 (rutin) 組合為EGCG-quercetin-rutin (EQR) 複方，以動物試驗探討EQR複方對於抗疲勞作用與調控腸道菌相之影響，並釐清其分子作用機轉。本研究由三大試驗組合而成，包括：跑步試驗、游泳試驗和抗疲勞作用。每一試驗皆使用8週齡雄性ICR小鼠，並隨機分成5組 (n=6/組)，分別有控制組 (control, C)、EQR複方低劑量組 (EQR formula-low dose, EQRF-LD)、EQR複方中劑量組 (EQR formula-medium dose, EQRF-MD)、EQR複方高劑量組 (EQR formula-high dose, EQRF-HD) 與正控制組 (positive control, PC)，為期4週。結果顯示，EQR複方可顯著增加ICR小鼠跑步時間與距離、游泳時間、運動後血漿乳酸清除率和葡萄糖含量、肝臟和腓腸肌肝醣含量，以及腓腸肌粒線體細胞色素c氧化酶活性，並顯著降低運動後血漿中氨和肌酸激酶含量。在基因表現結果顯示，EQR複方可顯著增加ICR小鼠肝臟中GLUT2、GK和PEPCK之基因表現。在糞便短鏈脂肪酸結果顯示，EQR複方可顯著增加ICR小鼠糞便中丁酸之含量。在腸道菌相結果顯示，EQR複方可顯著增加ICR小鼠腸道菌相多樣性，以及Lachnospiraceae、Coriobacteriaceae與Blautia之相對豐富度，並顯著降低Bacteroidaceae、Bacteroides、Clostridium、Parabacteroides、Parabacteroides goldsteinii與Bacteroides caccae之相對豐富度。綜合以上結果可知，EQR複方具有抗疲勞作用，其可透過增加能量物質之含量、清除或減少代謝產物之生成。另外，亦能藉由調節肝臟中碳水化合物代謝之基因表現，以及腸道菌相之組成，進而提升ICR小鼠之運動表現。因此，EQR複方未來可發展成具抗疲勞潛力之保健食品。

關鍵字

EQR複方 ； 抗疲勞 ； 跑步試驗 ； 游泳試驗 ； 腸道菌相 ； ICR小鼠

並列摘要

Fatigue is defined as full of fatigue, lack of mobility and inability to maintain voluntary physical activity, and it will have a negative impact on exercise intensity, work performance, family life, and social relationships. So far, there are many ways to prevent and improve fatigue, one of which is to take functional ingredient supplements. Based on previous findings, the composition of gut microbiota is related to the occurrence of fatigue, and a good composition of gut microbiota can enhance the exercise performance. Natural polyphenols have multiple functions, such as anti-oxidation, anti-inflammatory, anti-obesity, anti-cancer, etc., but the research on the anti-fatigue effect and gut microbiota of natural polyphenol compounds still remains unclear. Therefore, this study combined epigallocatechin gallate (EGCG), quercetin, and rutin, which are known to have an improvement effect on fatigue, as EGCG-quercetin-rutin (EQR) formula, and used animal experiments to explore anti-fatigue, regulation of gut microbiota and its molecular mechanisms of EQR formula. This study was composed of three major tests, including: running test, swimming test, and anti-fatigue effect. 8-week-old male ICR mice were used for each test and were randomly divided into 5 groups (n=6/group), including control (C), EQR formula-low dose (EQEF-LD), EQR formula-medium dose (EQRF-MD), EQR formula-high dose (EQRF-HD), and positive control (PC) groups for 4 weeks. The results showed that EQR formula significantly increased the running time and distance, swimming time, plasma clearance rate of lactate and glucose contents after exercise, hepatic and gastrocnemius glycogen contents, and gastrocnemius mitochondrial cytochrome c oxidase activity. EQR formula also significantly reduced plasma ammonia and creatine kinase contents after exercise in ICR mice. The gene expressions results showed that EQR formula significantly increased the gene expressions of GLUT2, GK, and PEPCK in liver of ICR mice. The results of fecal short-chain fatty acids showed that EQR formula significantly increased the contents of butyric acid in feces of ICR mice. The results of gut microbiota showed that EQR formula significantly increased the diversity of gut microbiota, and the relative abundance of Lachnospiraceae, Coriobacteriaceae, and Blautia in ICR mice. EQR formula also significantly reduced the relative abundance of Bacteroidaceae, Bacteroides, Clostridium, Parabacteroides, Parabacteroides goldsteinii, and Bacteroides caccae in ICR mice. Based on the above results, EQR formula had anti-fatigue functions, which increased the contents of energy substances, cleared or reduced the production of metabolites, and also regulatd the gene expressions of carbohydrate metabolism in the liver and the gut microbiota to improve the exercise performance of ICR mice. Therefore, the EQR formula can be developed into a functional foods with anti-fatigue potential in the future.

並列關鍵字

EQR formula ； anti-fatigue ； running test ； swimming test ； gut microbiota ； ICR mice

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