Ameliorative Effects of Glycine max on Metabolic Syndrome In Vitro and In Vivo
- 주제(키워드) Glycine max , metabolic syndrome
- 발행기관 고려대학교 대학원
- 지도교수 조홍연
- 발행년도 2014
- 학위수여년월 2014. 2
- 학위구분 석사
- 학과 일반대학원 식품생명공학과
- 원문페이지 123 p
- 실제URI http://www.dcollection.net/handler/korea/000000050061
- 본문언어 영어
- 제출원본 000045794149
초록/요약
Hypertension, diabetes and obesity are metabolic syndromes characterized by combination of medical disorder resulted from when occurring together. Metabolic syndrome is a cluster of metabolic risk factors. Metabolic disease are affected other metabolic disease each other and the chances of risk for metabolic disease such as cardiovascular problems are greater than any one disease presenting alone. Metabolic syndrome is connected to overweight, obesity, insulin resistance heart disease, genetic factor and other factors. This study was performed to investigate the ameliorative effect of Glycine max (soybean) on metabolic syndrome in ACE (angiotensin converting enzyme), differentiation of skeletal muscle cell line, human intestinal epithelial cell line and preadipocytes in vitro, and anti-obesity effect in C57BL/6N mice fed normal or high fat diet in vivo. In order to develop functional materials of the metabolic syndrome, the angiotensin converting enzyme inhibitory activity, inhibitors of glucose uptake and inhibitors of adipocytes differentiation were screened from germinated and proteolyzed soybean. Treatment of non-germinated, germinated and/or protease treated soy milk (0.1 mg/mL) among the extractions decreased adipocyte lipid accumulation in in vitr, in vivo, and glucose uptake activity using Caco-2 cell line and 2-NBDG. According to the results of in vitro assays, BWGSM6 (Bio-land protease treating whole germinated soy milk) was used to study inhibitory effect of soy milk on mRNA expression of major transcription factors related to cellular lipid accumulation using 3T3-L1 mature adipocytes. It was found that the mRNA expression of C/EBP α, PPAR γ, SREBP-1c, UCP-2, HSL and CPT-1. In addition to this study was also conducted to investigate the anti-obesity effect of soy milk on the high fat diet induced obese C57BL/6N mice in vivo. Experimental groups were normal diet group (ND), high fat diet group (HFD) and high fat diet + soy milk 5 % group (SMD) for 6 weeks. Consumption of soy milk effectively lowered body weight gain, food efficiency ratio, lipid accumulation in liver and relative epididymal fat mass compared with those of the high fat diet group. Also, LDL-cholesterol, triglyceride, total cholesterol, AST, ALT levels in the serum were significantly reduced by soy milk supplementation compared with those of the high fat diet group. Also, the soy milk increased serum HDL cholesterol level, which was decreased with the high fat diet. Liver and epididymal adipocyte tissue are found that the Mrna expression of C/EBP α, PPAR γ, SREBP-1c, UCP-2, HSL and CPT-1. In results, the soy milk might inhibit ACE, glucose uptake in Caco-2 cell line, preadipocyte differentiation, adipogenesis and lipogenesis in adipocyte cell line and high fat diet-induced obese mice.
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Contents
Abstract i
Contents iii
List of Figures viii
List of Tables xii
List of Schemes xiii
Ⅰ. Introduction 1
Ⅱ . Materials 9
1. Soybean samples 9
2. Chemicals 9
3. Cell line and medium 10
4. Animals 10
Ⅲ. Methods 11
1. Sample preparation 11
2. In vitro experiment 13
2.1. Porcine pancreatic lipase 13
2.2. Angiotensin I converting enzyme 14
2.3. L-6, Caco-2 cell culture 14
2.4. Glucose uptake assay 15
2.5. 3T3-L1 preadipocyte cell culture 17
2.6. Differentiation of 3T3-L1 preadipocyte 17
2.7. Measurement of 3T3-L1 preadipocyte cell viabiliy 18
2.8. Oil red O staining 20
2.9. RNA isolation 22
2.10. Reverse transcription-polymerase chain reaction analysis 23
3. In vivo experiment 25
3.1. Experimental design 25
3.1.1. Animals and diet 26
3.1.2. Blood sample and serum preparation 28
3.1.3. Sampling procedure of organs 29
3.2. Biochemical analysis 30
3.2.1. Analysis of Serum and Lipid content in Liver 30
3.2.2. Liver and epididymal adipose tissue histology and mean adipocyte surface area assays 31
3.3. Liver and epididymal adipose tissue gene expression 32
3.3.1. RNA isolation 32
3.3.2. Reverse transcription-polymerase chain reaction analysis 33
4. Statistical analyses 34
Ⅳ. Results and Discussion 35
1. In vitro studies 35
1.1. Screening of glucose uptake activity from soybean hot water extracts in L-6 cell line 35
1.2. Screening of glucose uptake activity from soybean hot water extracts in Caco-2 cell line 37
1.3. Screening of lipid accumulation from soybean hot water extracts in 3T3-L1 mature adipocytes. 39
1.4. Screening of glucose uptake activity from proteolyzed soybean extracts in L-6 cell line 41
1.5. Screening of glucose uptake activity from proteolyazed soybean extracts in Caco-2 cell line 43
1.6. Screening of lipid accumulation from proteolyzed soybean extracts in 3T3-L1 mature adipocytes 45
1.7. Screening of activity from soybean on porcine pancreatic lipase assay 47
1.8. Screening of inhibitory activity from soybean on ACE assay 49
1.9. Screening of glucose uptake activity from soybean extracts 51
1.10. Cell viability in 3T3-L1 preadipocyte 54
1.11. Morphological changes in 3T3-L1 differentiation 56
1.12. Changes of gene expression in 3T3-L1 cell by soybean extracts 59
2. In vivo studies 66
2.1. Body weight gain, food intake, and food efficiency ratio 66
2.2. Serum biochemical analysis 70
2.2.1. Serum lipid levels 70
2.2.2. Serum AST and ALT activities 71
2.3. Liver and epididymal adipose tissue weight, histology and adipocyte surface area 73
2.4. Hepatic lipid levels 78
2.5. Change of gene expression in C57BL/6N mice fed with soybean 81
2.5.1. Effect of soybean on mRNA expression of adipogenic and lipogenic transcription factors in epididymal adipose tissue 81
2.5.2. Effect of soybean on mRNA expression of adipogenic and lipogenic transcription factors in liver 88
Ⅴ. Conclusion 95
Ⅵ. References 97
Abstract in Korean
Acknowledgment in Korean
