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Metabolite profiling of biological specimens of mice fed high-fat diet depending on different rice cultivars using GC-TOF-MS

Metabolite profiling of biological specimens of mice fed high-fat diet depending on different rice cultivars using GC-TOF-MS
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대학원 식품공학과
이화여자대학교 대학원
Metabolite profiling of plasma in high-fat diet (HFD) induced obese mice was employed after being fed different rice cultivars such as anthocyanin-pigmented rice (BCR; Oryza sativa L. cv. Heugkwangbyeo) and high-amylose rice (GR; Oryza sativa L. cv. Goami3byeo). The plasma metabolites in mice rendered obese by intake of HFD were analyzed using gas chromatography-time of flight-mass spectrometry (GC-TOF-MS) and multivariate statistical analysis after being fed different diets for 8 weeks. The various diets were as follows 1) control, control rice (CR), black-colored rice (BCR), and garcinia cambogia extract (GCam); 2) control, control rice (CR), and high-amylose rice (GR). A total of twenty five metabolites such as 4 carbohydrates, 6 lipids, 5 organic acids, and 10 amino acids were identified in blood plasma. The group of mice fed BCR had significantly lower levels of glucose, galactose, propinoic acid, palmitic acid, strearic acid, and cholesterol compared with those fed CR after 8 weeks. In particular, the plasma level of phenylalanine, which is a biosynthetic precursor of anthocyanin, was markedly higher in the BCR group than in the CR group after 4 weeks. In addition, the GR was also effective in reducing the plasma levels of mannose, glucose, and cholesterol from 4 weeks to 8 weeks, compared to CR. In principal component analysis (PCA) plot, the BCR and GR group could be separated from the others after 4 weeks of the intervention. In terms of the major components contributing to the separation, mannose, isoleucine, threonine and phenylalanine were involved in BCR group, while propionic acid, valine, leucine, and proline were related to GR group, respectively. In partial least squares analysis (PLSR) in BCR group, body weight, glucose, and total cholesterol were positively correlated with the levels of propionic acid, butyric acid, cholesterol, alanine, ribitol, glucose, and galactose, whereas the high-density lipoprotein (HDL)/non-HDL ratio was negatively correlated with those of stearic acid, cholesterol, valine, leucine, glucose, and galactose. PLSR, antiobesity parameters including high-density lipoprotein (HDL)/non-HDL and HDL/total cholesterol (TC) ratio were found near GR group at an 8-week period in GR group. Propionic acid, proline, leucine, valine, and oxalic acid were negatively correlated with HDL/non-HDL ratio, whereas valine was negatively linked to HDL/TC ratio.;본 연구에서는 특수미를 섭취한 동물 생체시료(혈액) 내의 대사체 프로파일링 차이를 분석하였다. 동물실험은 고지방식이를 통해 비만이 유도된 쥐에 특수미를 투여하였다. 특수미의 종류는 총 2가지로 안토시아닌 색소가 풍부한 유색미인 흑광과 아밀로오스 함량이 높은 고아미3호를 사용하였다. 8주간 특수미를 섭취한 동물생체시료를 Gas chromatography time-of-flight mass spectrometry (GC-TOF-MS)와 다변량통계기법을 활용하여 대사체를 분석하였다. Target analysis 방법을 이용하여 생체시료 내 4가지의 당류, 6가지의 지방류, 5가지의 유기산류, 10가지의 아미노산류로 총 25가지의 대사산물을 동정하였다. 8주간 흑광을 투여한 군이 일반미를 투여한 군에 비해 glucose, galactose, propionic acid, palmitic acid, strearic acid, cholesterol 함량이 유의적으로 낮게 측정되었다. 특히 안토시아닌의 전구체로 알려진 phenylalanine은 4주간 흑광을 투여한 군이 일반미를 투여한 군에 비해서 유의적으로 높게 나타났다. 고아미3호를 투여한 군은 일반미를 투여한 군에 비해 mannose, glucose, cholesterol 함량이 4주 후부터 유의적으로 낮아지는 경향이 있었다. 주성분 분석에 따르면 4주 후부터 흑광과 고아미3호를 투여한 군이 일반미를 투여한 군으로부터 뚜렷하게 구분되었다. 이들 구분에 관여하는 주요 대사체들로는 흑광의 경우 mannose, isoleucine, threonine, phenylalanine이었으며, 고아미3호의 경우 propionic acid, valine, leucine, proline로 확인되었다. 이상의 결과를 종합해보면 두 가지의 특수미를 섭취한 생체시료에서 당류와 cholesterol의 함량이 일반미를 섭취한 군에 비해 유의적으로 낮게 측정되었고, 4주 후부터 다른 식이섭취 군으로부터 명확하게 구분되었다. 부분최소평방회귀분석에 본 데이터를 적용해본 결과 특수미를 섭취한 생체시료의 대사체인 Valine, Leucine은 HDL/non-HDL ratio와 부의 상관관계를 나타냈다.
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