심장 독성에서의 Sinapic Acid, Cucurbitacin I, 양배추 추출물의 억제 효과

목차 (Table of Contents)

• Part I Protective Effects of Sinapic Acid on Cardiac hyper- trophy as a cardiac toxicity in Neonatal Rat Cardio- myocytes 5
• 1.1 Introduction 6
• 1.1.1 Cardiac toxicity 6
• 1.1.2 Cardiac hypertrophy 8
• 1.1.3 Sirt3 and cardiac hypertrophy 11
• 1.1.4 SOD2 as a target protein of Sirt3 12
• 1.1.5 Sinapic acid 13
• 1.2 Materials and Methods 16
• 1.2.1 Cell culture and hypertrophic stimulation with PE 16
• 1.2.2 Cell viability assay 17
• 1.2.3 Immunostaining and cell size measurement 17
• 1.2.4 Quantitative real-time polymerase chain reaction (qRT-PCR) 18
• 1.2.5 Western blot analysis 19
• 1.2.6 Measurement of total ATP content 20
• 1.2.7 Transfection of small interfering RNA (siRNA) into cardiomyocytes and hypertrophic stimulation 20
• 1.2.8 Statistical analysis 21
• 1.3 Results 22
• 1.3.1 Cytotoxic actions of sinapic acid in cultured neonatal rat cardiom-yocytes 22
• 1.3.2 Sinapic acid attenuates hypertrophic responses in PE-stimulated cardiomyocytes 22
• 1.3.3 Sinapic acid inhibits MAPK signaling in PE-stimulated cardiomyo-cytes 23
• 1.3.4 Sinapic acid preserves mitochondrial integrity in PE-stimulated cardiomyocytes 24
• 1.3.5 Sinapic acid activates the SIRT3/SOD2 signaling pathway in PE-stimulated cardiomyocytes 25
• 1.3.6 The anti-hypertrophic impact of sinapic acid is attenuated in the SIRT3-silenced, hypertrophic cardiomyocytes 27
• 1.4 Discussion 29
• 1.5 Conclusion 32
• 1.6 References 33
• Part II Protective effects of cucurbitacin I on oxidative stress as a cardiac toxicity in H9C2 cardiomyoblasts 62
• 2.1 Introduction 63
• 2.1.1 Oxidative stress and cardiac toxicity 63
• 2.1.2 Cucurbitacin I and oxidative stress 65
• 2.2 Materials and Methods 67
• 2.2.1 Cell culture and oxidative stress induced by H2O2 67
• 2.2.2 Cell viability assay 67
• 2.2.3 Measurement of ROS production 68
• 2.2.4 Hoechst 33342 staining 68
• 2.2.5 Terminal deoxynucleotidyl transferase dUTP End Labeling (TUNEL) staining 69
• 2.2.6 Mitochondrial transmembrane potential (MMP) assessment 69
• 2.2.7 Western blot analysis 70
• 2.2.8 Quantitative real-time polymerase chain reaction (qRT-PCR) 71
• 2.2.9 Statistical analysis 72
• 2.3 Results 74
• 2.3.1 Cytotoxicity of Cu I in H9c2 cardiomyoblasts 74
• 2.3.2 Cu I rescues H9c2 cardiomyoblasts from H2O2-indcued oxidative stress 74
• 2.3.3 Cu I prevents the accumulation of ROS production in H2O2-treated H9c2 cardiomyoblasts 75
• 2.3.4 Cu I suppresses H2O2-induced apoptosis in H9c2 cardiomyo- blasts 76
• 2.3.5 Cu I prevents mitochondrial dysfunction upon H2O2-induced oxidative stress in H9c2 cardiomyoblasts 78
• 2.3.6 Cu I blocks the activation of MAPK signaling pathway in H2O2-treated H9c2 cardiomyoblasts 79
• 2.4 Discussion 80
• 2.5 Conclusion 83
• 2.6 References 84
• Part III Preventive effects of cabbage (Brassica oleracea var. capitata) extract on oxidative stress as a cardiac toxicity in H9c2 cardiomyoblasts 105
• 3.1 Introduction 106
• 3.2 Materials and Methods 109
• 3.2.1 Preparation of cabbage extract 109
• 3.2.2 Cell culture and induction of oxidative stress 109
• 3.2.3 Cell viability assay 110
• 3.2.4 Measurement of ROS production 110
• 3.2.5 Hoechst 33342 staining 111
• 3.2.6 Terminal deoxynucleotidyl transferase dUTP end labeling(TUNEL) staining 111
• 3.2.7 Mitochondrial transmembrane potential (MMP) assessment 111
• 3.2.8 Western blot analysis 112
• 3.2.9 Quantitative real-time polymerase chain reaction (qRT-PCR) 113
• 3.2.10 Statistical analysis 114
• 3.3 Results 116
• 3.3.1 Cytotoxicity of cabbage extract in H9c2 cardiomyoblasts 116
• 3.3.2 Cabbage extract protects H9c2 cardiomyoblasts against H2O2-induced oxidative stress 116
• 3.3.3 Cabbage extract inhibits H2O2-induced ROS production in H9c2 cardiomyoblasts 117
• 3.3.4 Cabbage extract rescues the antioxidant proteins in H2O2-treated H9c2 cardiomyoblasts 117
• 3.3.5 Cabbage extract blocks the MAPK signaling pathway in H2O2-treated H9c2 cardiomyoblasts 118
• 3.3.6 Cabbage extract suppresses H2O2-induced apoptosis in H9c2 cardiomyoblasts 119
• 3.3.7 Cabbage extract prevents mitochondrial dysfunction upon H2O2-induced oxidative stress in H9c2 cardiomyoblasts 121
• 3.4 Discussion 122
• 3.5 Conclusion 126
• 3.6 References 127
• Conclusion 147
• Abstract in Korean (국문 초록) 148
• Acknowledgement (감사의글) 151