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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 蔡沛學(Pei-Shiue Tsai) | |
| dc.contributor.author | Yu-Ting Hsu | en |
| dc.contributor.author | 許瑜庭 | zh_TW |
| dc.date.accessioned | 2021-06-07T17:40:41Z | - |
| dc.date.copyright | 2020-08-04 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-07-23 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/15446 | - |
| dc.description.abstract | 組織纖維化是因細胞外基質過度累積所造成的病症，此現象常出現於慢性疾病或慢性組織損傷患處。在人類腹膜透析病患中，腹膜纖維化會造成腹膜逐漸失去功能，不僅影響腹膜透析的效率，也可能演變成高死亡率之包囊性腹膜硬化症。纖維化相關蛋白TGF-β（transforming growth factor-β）為纖維化形成的主要調控因子，文獻指出TGF-β引發之NOX4酵素的過度活化亦參與了纖維化的形成，NOX（nicotinamide adenine dinucleotide phosphate oxidase）的主要功能之一為製造活性氧化物質（reactive oxygen species, ROS），且在腹膜透析病患中發現，腹膜間皮細胞產出之活性氧化物質是造成腹膜纖維化的主因之一。此篇論文將以人類腹膜間皮細胞（human peritoneal mesothelial cells, MeT-5A）及三品種豬隻 （Landrace × Yorkshire dam × Duroc sire, LYD）動物模式探討次氯酸鈉（NaOCl）引發腹膜纖維化之機制。實驗結果顯示，腹膜間皮細胞產生細胞內活性氧化物質的能力與總量與次氯酸鈉濃度及反應時間呈正相關，透過流式細胞儀的分析，腹膜間皮細胞經次氯酸鈉刺激後，會導致細胞壞死，而非細胞凋亡路徑的活化。此外，腹膜間皮細胞及豬隻腹膜纖維化模型研究結果顯示，次氯酸鈉可能藉由活化TGF-β1, NOX4, ERK引發腹膜纖維化。此篇論文中，我們亦探討間質幹細胞治療之應用，如生物材料包裹間質幹細胞，增加間質幹細胞之存活時間，及透過蛋白質體分析找出培養過間質幹細胞之培養液中抑制纖維化之相關分子，提供有潛力減緩纖維化及促進腹膜再生的研究方向。 | zh_TW |
| dc.description.abstract | Fibrosis, characterized by excessive accumulation of extracellular matrix, is a common pathological consequence in many chronic diseases or injuries occurring in various organs including in peritoneum. Patients who suffer from peritoneal fibrosis will gradually loss the normal function, and in some rare cases, develop encapsulat-ing peritoneal sclerosis which can result in high mortalities. TGF-β (transforming growth factor-β) has been identified as a major mediator in fibrotic process, and NOX4, a type of NADPH (nicotinamide adenine dinucleotide phosphate oxidase) oxidase, may have a pivotal role in mediating profibrotic responses. NOX-derived reactive oxygen species (reactive oxygen species, ROS) generation from peritoneal mesothelial cells is thought to be the key mechanism for TGF-β-induced peritoneal fibrosis in peritoneal dialysis patients. In this study, human peritoneal mesothelial cells (MeT-5A) and LYD (Landrace × Yorkshire dam × Duroc sire) porcine were used to investigate the mechanism of sodium hypochlorite (NaOCl)-induced perito-neal fibrosis. Intracellular ROS production in human mesothelial cells (MeT-5A) exhibited dose and time dependent increase under NaOCl stimulation. Besides, based on in vitro flow cytometry evidence, cell necrosis, but not apoptosis was the major cell fate of MeT-5A when treated with NaOCl. The protein expression of up-stream signals of fibrosis (TGF-β1, Nox4, and ERK) were also enhanced both in vitro and in vivo. Based on our in vitro and in vivo model systems, NaOCl induced TGF-β1/NOX4/ERK pathway which likely resulted in the peritoneal fibrosis ob-served in our porcine model. In this study, potential therapeutic options were also explored including biomaterial applications of MSC therapy and MSC cultured me-dium. Nevertheless, information derived from proteomic analysis would provide promising target molecules for the inhibition of peritoneal fibrosis and the regenera-tion of peritoneum. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-07T17:40:41Z (GMT). No. of bitstreams: 1 U0001-2207202015112600.pdf: 3475890 bytes, checksum: 05f7e4b79d98723f13c6d2d97029b754 (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 論文口試委員會審定書 i 誌謝 ii Abstract iii 中文摘要 v Contents vii List of figures xi List of tables xiii Chapter 1 Introduction 1.1 The structure and physiological functions of peritoneum 1 1.2 Long-term peritoneal dialysis (PD) induced peritoneal fibrosis (PF) in PD patients 1.2.1 The pathological structure of peritoneum upon fibrosis 2 1.2.2 The molecular mechanism of peritoneal fibrosis induced by peritoneal dialy-sis 3 1.2.3 Reactive oxygen species (ROS) and associated molecules as candidate markers of peritoneal dialysis-induced peritoneal fibrosis 5 1.3 Potential mechanisms of TGF-β-induced upregulation of NOX4 expression 5 1.4 The features of different peritoneal fibrosis animal models 7 1.5 Potential therapies for peritoneal fibrosis by mesenchymal stem cells (MSC) 1.5.1 The functions of Mesenchymal stem cells (MSC) 9 1.5.2 The inhibition efficacy of fibrosis in MSC therapies 10 1.5.3 The approaches of MSC therapies 11 1.6 Aims of this project 14 Chapter 2 Materials and Methods 2.1 Cell culture and cell lysate preparation 15 2.2 Apoptosis and necrosis detection by flow cytometry 16 2.3 Cell viability assay (MTT assay) 17 2.4 Cellular ROS detection assay 17 2.5 Establishment of sodium hypochlorite-induced peritoneal fibrosis and time point sample collections 18 2.6 Western blot 19 2.7 Immunofluorescence assay (IFA) 21 2.8 Cell sheet culture system 21 2.9 Mesenchymal Stem Cell (MSC) culture in gelatin-based hydrogel 23 2.10 Cell viability of MSC in hydrogel 23 2.11 Proteomic analyses of human umbilical cord-derived stem cell culture su-pernatant 24 2.12 Statistical analysis 25 Chapter 3 Results 3.1 NaOCl promoted generation of intracellular ROS in MeT-5A 26 3.2 NaOCl promoted mesothelial cell death via necrosis 29 3.3 NaOCl-induced TGF-β1, Nox4, and p-ERK expressions in mesothelial cells 31 3.4 NaOCl induced overexpression of TGF-β1 on the peritoneum 33 3.5 NOX4 protein expression was increased after 0.1% NaOCl injury 35 3.6 NaOCl promoted overexpression of p-ERK in abdominal wall, a downstream signaling molecule of TGF-β-NOX4 axis 37 3.7 Myofibroblast marker α-SMA protein expression showed time-dependent in-crease in 0.1% NaOCl-induced fibrosis model 39 3.8 Type I collagen deposition was increased in 0.1% NaOCl-induced peritoneal fibrosis pig model 41 3.9 Mesenchymal stem cells as therapeutic approach 43 3.10 Gelatin-based hydrogel as bio-compatible material for MSCs therapy model 45 3.11 Proteomic analysis of MSC culture supernatant 47 Chapter 4 Discussion 50 Chapter 5 Conclusion and future work 61 Reference 63 | |
| dc.language.iso | en | |
| dc.subject | 間質幹細胞治療 | zh_TW |
| dc.subject | 腹膜纖維化 | zh_TW |
| dc.subject | TGF-β1 | zh_TW |
| dc.subject | NOX4 | zh_TW |
| dc.subject | ERK | zh_TW |
| dc.subject | ERK | en |
| dc.subject | NOX4 | en |
| dc.subject | TGF-β1 | en |
| dc.subject | MSC therapy | en |
| dc.subject | peritoneal fibrosis | en |
| dc.title | 以豬隻動物模式探討次氯酸鈉引發腹膜纖維化之機制 | zh_TW |
| dc.title | The Mechanism of Sodium Hypochlorite-induced Porcine Model of Peritoneal Fibrosis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林水龍(Shuei-Liong Lin),蔡素宜(Su-Yi Tsai),李雅珍(Ya-Jane Lee),陳怡婷(Yi-Ting Chen) | |
| dc.subject.keyword | 腹膜纖維化,TGF-β1,NOX4,ERK,間質幹細胞治療, | zh_TW |
| dc.subject.keyword | peritoneal fibrosis,TGF-β1,NOX4,ERK,MSC therapy, | en |
| dc.relation.page | 69 | |
| dc.identifier.doi | 10.6342/NTU202001737 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2020-07-23 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
| 顯示於系所單位： | 獸醫學系 | |
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