Title Page
Contents
Part Ⅰ. Amelioratory effects of hydroferulic acid from Momordica charantia leaf on oxidative stress and liver pathology in mice with non-alcoholic steatohepatitis 5
CONTENTS 5
Abstract 9
Chapter 1. Introduction 11
Chapter 2. Materials and Methods 15
2.1. Materials and Methods 15
2.2. Biochemical Analysis 17
2.3. Histological Analysis 17
2.4. mRNA Expression Analysis 19
2.5. α-SMA, F4/80 and CD68 Immunostaining 19
2.6. Protein Expression Analysis 20
2.7. Statistical Analysis 20
Chapter 3. Results 21
3.1. Effect of HFA on liver coefficients in WD-induced NASH mice 21
3.2. Effect of HFA on liver histopathology in WD-induced NASH mice 29
3.3. Serum biochemical effects of HFA on WD-induced NASH mice 36
3.4. Effect of HFA on hepatic lipids in WD-induced NASH mice 39
3.5. Effects of HFA on liver fibrosis and inflammation-related gene expression in NASH mice 42
Chapter 4. Discussion 57
Chapter 5. Conclusions 63
References[원문불량;p.56,60-62,65] 64
Abstract (in Korean) 74
Part Ⅱ. Viscoelastic behavior and structural property of Momordica charantia leaf powder-corn starch blends as food inks for 3D printer 76
CONTENTS 76
Abstract 81
Chapter 1. Introduction 83
Chapter 2. Materials and Methods 85
2.1. Materials 85
2.2. Food ink mixture preparation 86
2.3. Scanning electron microscopy (SEM) 88
2.4. Dynamic viscoelastic properties of 3D printing food inks 88
2.5. FTIR spectrum of 3D printing food inks 89
2.6. Texture properties of 3D printing food inks 89
2.7. Printability assessment of 3D printing food inks 90
2.8. International Dysphagia Diet Standardization Initiative(IDDSI) test of 3D food inks 91
2.9. Statistical analysis 92
Chapter 3. Results 93
3.1. Particle size of MCLP 93
3.2. Rheological characteristics of 3D printing food inks 95
3.3. Gel properties of 3D printing food inks 98
3.4. FT-IR absorption characteristics of 3D printing food inks 101
3.5. Printing performance of 3D printing food inks 103
3.6. Scanning electron microscopy (SEM) of 3D printing food inks 106
3.7. IDDSI test of 3D printing food inks 110
Chapter 4. Discussion 113
Chapter 5. Conclusions 118
References 119
Abstract (in Korean) 125
Part Ⅲ. Momordica charantia leaf powder-corn starch 3D food ink as functional foods in controlling gestational hyperglycemia in obese mice fed with high-fat diet 126
CONTENTS 127
Abstract 129
Chapter 1. Introduction 131
Chapter 2. Materials and Methods 134
2.1. Materials and Methods 134
2.2. Biochemical analysis 136
2.3. Histological analysis 136
2.4. Analysis of mRNA expression 137
2.5. Analysis of protein expression 138
2.6. Statistical analysis 139
Chapter 3. Results 140
3.1. Effect of MCIM on histopathology in HFD-induced GDM mice 140
3.2. Effect of MCIM on OGTT and ITT in HFD-induced GDM mice 146
3.3. Serum biochemical effects of MCIM on HFD-induced GDM mice 149
3.4. Effect of MCIM on hepatic lipids in HFD-induced GDM mice 152
3.5. Effect of MCIM on PI3K pathway in HFD-induced GDM mice 155
Chapter 4. Discussion 160
Chapter 5. Conclusions 166
References[원문불량;p.166] 167
Abstract (in Korean) 175
Part Ⅱ 79
Table 1. Composition of different inks. 87
Table 2. Classification of samples based on the IDDSI test. 111
Part Ⅰ 7
Figure 1. Gross morphological examination and respective histopathological microphotographs of the liver. Normal chow-fed mice (CON); NASH treated with... 22
Figure 2. Histological examination of liver sections stained with H&E (200×magnification). Normal chow-fed mice (CON); NASH treated with saline... 24
Figure 3. Histological examination of liver sections stained with H&E (400×magnification). Normal chow-fed mice (CON); NASH treated with saline (NASH);... 25
Figure 4. Effect of HFA on liver coefficients in WD-induced NASH mice liver injury. Body weight (A) and liver weight (B). Normal chow-fed mice (CON);... 27
Figure 5. Effect of HFA on liver coefficients in WD-induced NASH mice. Liver to body weight (LBW) ratio. Normal chow-fed mice (CON); NASH treated with... 28
Figure 6. Oil Red O staining to visualize lipid droplets of hepatic sections in WD-induced NASH mice. Normal chow-fed mice (CON); NASH treated with... 30
Figure 7. Masson's trichrome staining to visualize fibrotic changes of hepatic sections in WD-induced NASH mice. Normal chow-fed mice (CON); NASH... 32
Figure 8. Sirius red staining to visualize fibrotic changes of hepatic sections in WD-induced NASH mice. Normal chow-fed mice (CON); NASH treated with... 33
Figure 9. Histology score for steatosis, lobular inflammation, ballooning and NAFLD Activity Score were quantified. Normal chow-fed mice (CON); NASH... 35
Figure 10. Serum levels of triglyceride (TG) and free fatty acids (FFAs) in NASH mice. A. Serum TG. B. Serum FFAs. Normal chow-fed mice (CON);... 37
Figure 11. Serum levels of cholesterol (TC) and low-density lipoprotein/very low-density lipoprotein (LDL/VLDL) in NASH mice. A. Serum TC. B. Serum... 38
Figure 12. Liver tissue levels of triglyceride (TG) and free fatty acids (FFAs) in NASH mice. A. Liver tissue TG. B. Liver tissue FFAs. Normal chow-fed mice... 40
Figure 13. Liver tissue levels of low-density lipoprotein/very low-density lipoprotein (LDL/VLDL) and High-density lipoprotein (HDL) in NASH mice.... 41
Figure 14. Immunohistochemistry for α-smooth muscle actin (α-SMA) in NASH mice. A. immunohistochemistry shows the expression of α-SMA. Scale bar, 50... 43
Figure 15. The expression of α-SMA and Col1a1 during liver fibrosis in NASH mice. A. Western blot analysis of α-SMA and Col1a1 with GAPDH as the... 44
Figure 16. The expression of Col1a1 and Col1a2 during liver fibrosis in NASH mice. The RNA expression levels of A. Col1a1, B. Col1a2. GAPDH was used as... 45
Figure 17. Immunohistochemistry for F4/80 in NASH mice. A. immunohistochemistry shows the expression of F4/80. Scale bar, 50 μm. B.... 47
Figure 18. Immunohistochemistry for CD68 in NASH mice. A. immunohistochemistry shows the expression of CD68. Scale bar, 50 μm. B.... 48
Figure 19. The expression of IL-1β and TNF-α in NASH mice. The RNA expression levels of A. IL-1β, B. TNF-α. GAPDH was used as an internal control.... 49
Figure 20. The expression of MCP-1 in NASH mice. The RNA expression levels of MCP-1. GAPDH was used as an internal control. Normal chow-fed mice... 50
Figure 21. The expression of MerTK pathday in NASH mice (1). A. Western blot analysis of MerTK pathday with GAPDH as the loading control. B. Gas6... 52
Figure 22. The expression of MerTK pathday in NASH mice (2). A. MerTK relative portein density. B. p-ERK/ERK relative portein density. Normal chow-... 53
Figure 23. The expression of MerTK pathday in NASH mice (3). TGF-β relative portein density. Normal chow-fed mice (CON); NASH treated with saline... 54
Figure 24. Immunohistochemistry for MerTK in NASH mice. Immunohistochemistry shows the expression of MerTK. Scale bar, 100 μm.... 55
Figure 25. The expression of TGF-β1 and TGF-β2 in NASH mice. The RNA expression levels of A. TGF-β1, B. TGF-β2. GAPDH was used as an internal... 56
Part Ⅱ 80
Figure 1. SEM images for MCL powder samples. Particle size: A.75 μm, B.131 μm and C.200 μm. 94
Figure 2. Rheological properties of ink mixtures at various concentrations (1). A. Viscosity versus angular ffrequency relationships for ink mixtures. B. Elastic... 96
Figure 3. Rheological properties of ink mixtures at various concentrations (2). A. Viscous modulus (G″) versus angular ffrequency relationships for ink... 97
Figure 4. Texture properties of different ink mixtures (1). A. Hardness. B. Adhesiveness. Data are the geometric mean ± s.d. n=6. Statistical analysis was... 99
Figure 5. Texture properties of different ink mixtures (2). A. Chewiness. B. Gumminess. Data are the geometric mean ± s.d. n=6. Statistical analysis was... 100
Figure 6. FTIR absorption spectra of different ink mixtures. 102
Figure 7. 3D printing performance of different ink mixtures (1). A. Printed samples using 75μm MCLP particle size inks. B. Printed samples using 75μm... 104
Figure 8. 3D printing performance of different ink mixtures (1). A. Printed samples using 200μm MCLP particle size inks. B. Representative images of 3D... 105
Figure 9. Microstructure of different ink formulations (1). The magnifications of each group images are set as 500×. A. 5% 75 μm MCLP with 15% corn starch.... 107
Figure 10. Microstructure of different ink formulations (2). The magnifications of each group images are set as 500×. A. 10% 75 μm MCLP with 10% corn... 108
Figure 11. Microstructure of different ink formulations (3). The magnifications of each group images are set as 500×. A. 15% 75 μm MCLP with 5% corn starch.... 109
Part Ⅲ 128
Figure 1. Histological examination of epididymis adipose tissue sections stained with H&E in HFD-induced GDM mice. Normal chow-fed mice (CON); high fat... 141
Figure 2. Histological examination of liver tissue sections stained with H&E in HFD-induced GDM mice. Normal chow-fed mice (CON); high fat diet treated... 143
Figure 3. Histological examination of pancreas tissue sections stained with H&E in HFD-induced GDM mice. Normal chow-fed mice (CON); high fat diet treated... 145
Figure 4. Effect of MCIM on OGTT in HFD-induced GDM mice. A. Oral glucose tolerance test (OGTT). B. Area under the curve of the OGTT. Normal... 147
Figure 5. Effect of MCIM on ITT in HFD-induced GDM mice. A. Insulin tolerance test (ITT). B. Area under the curve of the ITT. Normal chow-fed mice... 148
Figure 6. Serum levels of triglyceride (TG) and free fatty acids (FFAs) in NASH mice. A. Serum TG. B. Serum FFAs. Normal chow-fed mice (CON); high fat... 150
Figure 7. Serum levels of cholesterol (TC) and low-density lipoprotein/very low-density lipoprotein (LDL/VLDL) in NASH mice. A. Serum TC. B. Serum... 151
Figure 8. Liver tissue levels of triglyceride (TG) and free fatty acids (FFAs) in NASH mice. A. Liver tissue TG. B. Liver tissue FFAs. Normal chow-fed mice... 153
Figure 9. Liver tissue levels of low-density lipoprotein/very low-density lipoprotein (LDL/VLDL) and High-density lipoprotein (HDL) in NASH mice.... 154
Figure 10. Immunohistochemistry for Insulin in NASH mice. Immunohistochemistry shows the expression of Insulin. Scale bar, 50 μm.... 156
Figure 11. The expression of glucagon during liver fibrosis in NASH mice. A. Western blot analysis of glucagon with GAPDH as the loading control. B.... 157
Figure 12. The expression of PI3K pathday in NASH mice (1). A. Western blot analysis of PI3K pathday with GAPDH as the loading control. B. PI3K relative... 158
Figure 13. The expression of PI3K pathday in NASH mice (2). A. p-AKT relative portein density. B. GLUT4 relative portein density. Normal chow-fed... 159