| 摘要: | 生物體內代謝物是生理反應的最終產物，藉由代謝體學能夠提供和表現型有最直接關聯的代謝物變化資訊。本研究以標的以及非標的代謝體分析方法研究植物化合物，以及植物成分與CoQ10影響的代謝物改變，並結合腸道菌相分析探討CoQ10對腸道菌的影響。在植物化合物的部分，以活性追蹤策略進行植物成分的分離純化，從沉香葉中鑑定出對MMP-2及-9具有正向調節活性的二次代謝物PA，並發現UV照射會改變其部分結構的位向，且可能因此改變其活性。同時運用傳統分離純化分析以及LC-MS非標的代謝體分析方法，本研究發現芝麻渣中被BSFL主要利用的成分為DG，且有數種胺基酸、醣類以及脂質在不同生長期BSFL蟲體與糞便中有顯著的差異變化，並推測部分代謝物是由幼蟲體內共生菌代謝生成。利用2D-HPLC系統，本研究進行了標的代謝物—Phe鏡相異構物在人類、大鼠、小鼠血液及尿液中的定量方法開發，且分析發現在DAO缺乏的哺乳動物體內D-Phe的含量顯著上升。CoQ10是一種常見的健康食品，在粒線體內參與能量的代謝，並且被認為能促進健康以及改善許多疾病，如代謝疾病、神經疾病，以及心血管疾病。為了了解CoQ10作為健康食品對動物體的影響，本研究以100 mg/ kg CoQ10餵食健康大鼠，分析大鼠體內代謝物以及糞便中的腸道菌變化。將已開發的Phe鏡像異構物分析方法運用於CoQ10代謝物分析，結果顯示CoQ10會使大鼠尿液中%D-Phe顯著上升。使用LC-MS非標的代謝體分析以及NMR標的代謝體分析方法，結合IPA代謝路徑分析，結果顯示CoQ10會導致大鼠血漿中多種小分子代謝物與脂蛋白的含量變化，這些差異性代謝物的變化可能來自於碳水化合物以及脂質代謝的改變。透過16S rRNA長片段基因分析，本研究發現CoQ10會影響糞便中數種腸道菌的組成。透過標的與非標的代謝體學研究方法，本研究分析得到了數種差異性代謝物，並討論其變化相關的生物功能。
In living organisms, metabolites are the final products of physiological reactions. Through metabolomic studies, the metabolites changes that are most directly related to the phenotypes can be explored. In this research, targeted and untargeted metabolomic analysis methods were used to study phytochemicals and metabolomes that are affected by the supplementation of phytochemicals or CoQ10. Additionally, the gut microbiota analysis was performed to explore the effect of CoQ10 on gut bacteria. In the study of phytochemicals, bioassay-guided strategy was applied for compound separation, and PA was purified and identified from Aquilaria sinensis leaves. Up regulatory activity on MMP-2 and -9 was found on PA, however, its structure could be changed by UV irradiation and may therefore alter its bioactivity. By using traditional compound separation and LC-MS based untargeted metabolomic analysis, DG was found to be the major component in sesame residues utilized by BSFL, and several amino acids, sugars, and lipids were found to be the differential components among different growth stages of BSFL and feces. Additionally, some of the metabolites are found potentially been metabolized by symbiotic bacteria in BSFL. In the section of targeted chiral metabolite analysis, a 2D-HPLC method for the determination of Phe enantiomers in the plasma and urine of humans, rats, and mice has been developed. The results showed that D-Phe contents were highly correlated to DAO activity. CoQ10 is widely used as a healthy food supplement, and is considered to improve health conditions and many diseases such as metabolic syndromes, neurological diseases, and cardiovascular diseases. In order to understand the effect of CoQ10 on healthy animals, healthy SD rats were fed with 100 mg/kg CoQ10, and the changes of metabolites and gut bacteria were analyzed. The developed Phe enantiomer analysis method was applied to this study, and the result showed that %D-Phe significantly increased in the urine. By using LC-MS based untargeted metabolomic analysis and NMR based targeted metabolomic analysis, the relative contents of various small molecule metabolites and lipoproteins were found altered in the plasma. Through the IPA pathway analysis, the metabolite changes were supposed to correlate to carbohydrate and lipid metabolism. Long sequence 16S rRNA gene sequencing was performed on the feces samples, and the results showed that CoQ10 could affect the composition of several gut bacteria in feces. Through targeted and untargeted metabolomic research methods, several differential metabolites were discovered in this study, and the biological functions related to their changes were discussed. |
