博碩士論文 102284004 詳細資訊




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姓名 蕭安淇(An-Ci Siao)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 內皮素誘導前脂肪細胞生長的訊息路徑
(Signal pathways of endothelins in inducing preadipocyte growth)
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摘要(中) 肥胖與糖尿病、高血壓、癌症和心血管疾病的風險有關,其特徵在於增加脂肪細胞的有絲分裂與脂肪的形成。然而,脂肪細胞的有絲分裂和脂肪形成是可以通過內分泌、遺傳、生長因子和營養來調節。本論文的總體目標主旨是在了解內皮素 (ET) 訊息途徑在控制前脂肪細胞有絲分裂作用與綠茶表沒食子兒茶素沒食子酸酯 (EGCG;主要兒茶素)、胰島素 (INS) 和類胰島素生長因子 (IGFs) 之間的關聯。這項研究的結果 (第一章) 表明第一型內皮素是透過ETAR、PKC、STAT3、AMPK、c-JUN、ERK、鞘氨醇激酶和鞘磷脂酶途徑刺激3T3-L1前脂肪細胞的生長。我們發現第三型內皮素 (第二章) 透過ETAR、AMPK、JNK/c-JUN途徑,而不是ERK與PKC途徑去刺激3T3-L1前脂肪細胞生長。這結果顯示,第一型內皮素與第三型內皮素會透過不同的訊息途徑去刺激前脂肪細胞的增生。第一型內皮素 (第三章) 可能透過ETAR、ETBR、PKC、STAT3和AMPK途徑去影響人類白色前脂肪細胞的生長。結果顯示EGCG (第四章) 透過ERK、c-Jun和SATA3途徑去抑制第一型內皮素所誘導的前脂肪細胞的生長。有趣的是我們也發現沒食子酸 (gallic acid) 與兒茶素 (catechin) 抑制第一型內皮素誘導的前脂肪細胞的生長,而表兒茶素 (epicatechin) 或表沒食子兒茶素 (epigallocatechin) 或表兒茶素沒食子酸酯 (epicatechin gallate) 則沒有這樣的作用,研究結果證明綠茶中的兒茶素具有特異性的調節作用。內皮素、胰島素或類胰島素生長因子對前脂肪細胞生長的交互作用中 (第五章),結果表明第一型內皮素和胰島素會透過ERK途徑的協同作用來增強前脂肪細胞生長,而不是透過p38或JNK途徑。我們觀察到第二型內皮素或第三型內皮素與胰島素刺激前脂肪細胞生長具有相似的協同作用。此外,第一型內皮素、第二型內皮素、第三型內皮素都會增強類胰島素生長因子所影響的3T3-L1前脂肪細胞的生長。本文的研究結果清楚地描述各種內皮素荷爾蒙調控白色前脂肪細胞生長會透過不同的訊號途徑,並可能提供內皮素、EGCG、胰島素和類胰島素生長因子介導前脂肪細胞生長相互作用的機制,進而導致脂肪細胞活性的變化和調節脂肪細胞相關的肥胖症和其他生物醫學疾病。
摘要(英) Obesity is associated with the risks of diabetes, hypertension, cancer, and cardiovascular diseases and is characterized with increased mitogenesis and adipogenesis of fat cells. In turn, adipocyte mitogenesis and adipogenesis can be regulated by endocrine, genetic, growth factors, and nutritional cues. The overall objective of the dissertation was designed to understand the endothelin (ET) signaling pathways in controlling preadipocyte mitogenesis in relation to green tea epigallocatechin gallate (EGCG; the major catechin), insulin (INS), and insulin-like growth factors (IGFs). The results of this study (Chapter One) indicated that ET-1 stimulated 3T3-L1 preadipocyte growth via the ETAR, PKC, STAT3, AMPK, c-JUN, ERK, sphingosine kinase, and sphingomyelinase pathways. ET-3 (Chapter Two) was found to stimulate 3T3-L1 preadipocyte growth via the ETAR, AMPK, JNK/c-JUN, but not ERK or PKC, pathways. This suggests that ET-3 exhibits somewhat different signals from ET-1 to stimulate preadipocyte proliferation. ET-1 (Chapter Three) was found to stimulate human primary preadipocyte growth possibly via the ETAR, ETBR, PKC, STAT3, and AMPK pathways. Interestingly, neither ET-2 nor ET-3 altered the growth of human white preadipocyte. EGCG (Chapter Four) was found to suppress ET-1-induced growth of 3T3-L1 preadipocytes though the ERK, c-JUN, and STAT3 pathways. Interestingly, gallic acid, catechin, but not epicatechin, or epigallocatechin, or epicatechin gallate, were also found to inhibit ET-1-induced growth of preadipocytes, suggesting the catechin-specific effect of green tea. The interactive effect of ET with INS or IGF on preadipocyte growth (Chapter Five) indicated that ET-1 and insulin had a synergistic effect on preadipocyte growth via the ERK but not p38 or JNK pathway. A similar synergistic effect of either ET-2 or ET-3 with INS to stimulate preadipocyte growth was observed. In addition, ET-1, ET-2, or ET-3 generally enhanced IGF-stimulated growth of 3T3-L1 preadipocytes. The results of this dissertation delineate distinct signal pathways of various ET hormones in controlling white preadipocyte growth and may provide a mechanism by which ETs interact with EGCG, INS, and IGF to mediate preadipocyte growth and thereby leading to changes in fat cell activity and coordinating fat cell-related obesity and other biomedical diseases.
關鍵字(中) ★ 第一型內皮素
★ 第三型內皮素
★ 胰島素
★ 類胰島素生長因子
★ 前脂肪細胞增生
★ 表沒食子兒茶素沒食子酸酯
關鍵字(英) ★ Endothelin-1
★ Endothelin-3
★ Insulin
★ insulin-like growth factors
★ Preadipocyte growth
★ Epigallocatechin gallate
論文目次 Chinese Abstract i
English Abstract ii
Declaration iv
Acknowledgments v
Table of Contents vi
List of Figures ix
List of Appendix Figures xii
Abbreviation xiii
Overall introduction 1
Chapter I – Endothelin-1 stimulates preadipocyte growth 4
I-1. Introduction 6
I-2. Materials and Methods 9
I-2.1 Chemical reagents 9
I-2.2 Cell culture 9
I-2.3 Growth stimulation experiments 10
I-2.4 Inhibitor experiments 11
I-2.5 Primary cell culture 12
I-2.6 Western blot analysis 13
I-2.7 Statistical analysis 13
I-3. Results 15
I-3.1 Effect of ET-1 on preadipocyte growth 15
I-3.2 ET-1 increases preadipocyte growth through ETAR but not ETBR 15
I-3.3 ERK and JNK are involved in the ET-1-stimulated proliferation of preadipocytes 16
I-3.4 PI3K/AKT inhibitor suppresses ET-1-induced stimulation of preadipocyte growth 18
I-3.5 JAK2/STAT3 inhibitor suppresses the ET-1 stimulation of preadipocyte growth 18
I-3.6 AMPK inhibitor suppresses the ET-1 stimulation of preadipocyte growth 19
I-3.7 PKC inhibitor suppresses the ET-1 stimulation of preadipocyte growth 20
I-3.8 SphK and SMase2 inhibitors suppress the ET-1 stimulation of preadipocyte growth 21
I-3.9 Cell type dependence of ET-1 23
I-4. Discussion 23
I-5. References 28
Chapter II – Endothelin-3 stimulates preadipocyte growth 34
II-1. Introduction 36
II-2. Materials and Methods 39
II-2.1 Chemical reagents 39
II-2.2 Cell culture 39
II-2.3 Growth stimulation experiments 39
II-2.4 Inhibitor experiments 40
II-2.5 Western blot analysis 40
II-2.6 Statistical analysis 41
II-3. Results 41
II-3.1 Effect of ET-3 on preadipocyte growth via ETAR but not ETBR 41
II-3.2 Effect of ET-3 on preadipocyte growth through ERK and JNK 42
II-3.3 Effect of ET-3 on preadipocyte growth are involved in ERK and JNK 42
II-3.4 Effect of ET-3 on preadipocyte growth via the SphK and SMase2 pathways 43
II-3.5 Cell type dependence of ET-3 44
II-4. Discussion 44
II-5. References 47
Chapter III – Endothelin-1 stimulates human preadipocyte growth 54
III-1. Introduction 56
III-2. Materials and Methods 58
III-2.1 Chemical reagents 58
III-2.2 Cell culture 58
III-2.3 Growth stimulation experiments 59
III-2.4 Western blot analysis 59
III-2.5 Statistical analysis 60
III-3. Results 61
III-3.1 ET-1 stimulated the growth of HWP-visceral cell proliferation 61
III-3.2 ET-1 increased HWP-subcutaneous cell proliferation 61
III-3.3 ET-1-stimulated HWP-visceral growth via the ETAR pathway 61
III-3.4 Different effects of various endothelin hormones in HWP-visceral cell 62
III-4. Discussion 63
III-5. References 65
Chapter IV – Green tea epigallocatechin gallate suppressed ET-1 stimulation of 3T3-L1 preadipocyte growth through the ERK, cJUN, and STAT3 pathways. 70
IV-1. Introduction 72
IV-2. Materials and Methods 74
IV-2.1 Chemical and reagents 74
IV-2.2 Cell culture 74
IV-2.3 Growth stimulation experiments 75
IV-2.4 MTT assay 76
IV-2.5 Western blot analysis 76
IV-2.6 Statistical analysis 77
IV-3. Results 78
IV-3.1 EGCG suppressed ET-1- stimulated proliferation of preadipocytes. 78
IV-3.2 EGCG suppressed ET-1-induced increase in the phosphorylation of MAPK proteins. 78
IV-3.3 EGCG suppressed ET-1-induced increase in the phosphorylation of STAT3 but not AMPK or PKC proteins. 79
IV-3.4 Other green tea catechins altered ET-1-stimulated preadipocyte growth. 79
IV-4. Discussion 81
IV-5. References 82
Chapter V - The interaction effect of endothelins with insulin and insulin-like growth factors to stimulate 3T3-L1 preadipocyte growth 92
V-1. Introduction 94
V-2. Materials and Methods 95
V-2.1 Chemical and reagents 95
V-2.2 Cell culture 95
V-2.3 Cell proliferation experiment 96
V-2.4 MTT assay 96
V-2.5 Statistical analysis 97
V-3. Results 98
V-3.1 ET-1 synergized with INS to stimulate preadipocyte growth 98
V-3.2 The ERK pathway mediated the synergistic effect of ET-1 with INS to stimulate mitogenesis of preadipocytes 98
V-3.3 ET-1 synergized with IGF to stimulate preadipocyte growth 99
V-3.4 ET-2 synergized with INS and IGF to stimulate preadipocyte growth 99
V-3.5 ET-3 synergized with INS and IGF to stimulate preadipocyte growth 100
V-4. Discussion 101
V-5. References 103
Conclusions 111
References 113
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指導教授 高永旭(Yung-Hsi Kao) 審核日期 2020-11-27
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