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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 江伯倫(Bor-Luen Chiang) | |
dc.contributor.author | Sheng-Chieh Lin | en |
dc.contributor.author | 林聖傑 | zh_TW |
dc.date.accessioned | 2021-06-08T03:41:22Z | - |
dc.date.copyright | 2019-08-26 | |
dc.date.issued | 2019 | |
dc.date.submitted | 2019-06-27 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21656 | - |
dc.description.abstract | 氣喘是兒童常見的發炎性呼吸道疾病。胸腺基質淋巴生成素是一種會引起免疫作用進而引起發炎反應的細胞激素。胸腺基質淋巴生成素和胸腺基質淋巴生成素接受器,目前在氣喘患者的致病機轉中扮演著重要的角色,但是否為氣喘的生物指標尚不清楚。抗胸腺基質淋巴生成素抗體在氣喘氣道重塑與對基質金屬蛋白酶、結締組織生長因子和轉化生長因子-β的影響,以及基質金屬蛋白酶和結締組織生長因子在氣喘氣道重塑中的相關性也值得研究與討論。因此在體外的研究中,我們嘗試研究氣喘和非氣喘兒童的胸腺基質淋巴生成素和胸腺基質淋巴生成素接受器的表現,我們招募了氣喘和非氣喘的兒童並記錄了臨床症狀。我們抽取血液以檢查胸腺基質淋巴生成素,並使用逆轉錄聚合鏈反應和即時聚合酶鏈鎖反應測量胸腺基質淋巴生成素接受器基因的表現。在體內的研究中,我們建立氣喘小鼠模型以評估結締組織生長因子和基質金屬蛋白酶在氣道重塑中的表現並且研究抗胸腺基質淋巴生成素抗體對氣喘氣道重塑的結構變化,同時研究抗胸腺基質淋巴生成素抗體對基質金屬蛋白酶、結締組織生長因子和轉化生長因子-β的影響。在我們對氣喘兒童的研究結果中,胸腺基質淋巴生成素和胸腺基質淋巴生成素接受器在氣喘兒童中顯著高於非氣喘的兒童。在我們對氣喘小鼠氣道重塑的研究結果中,發現結締組織生長因子有顯著的增加並且與基質金屬蛋白酶-9呈現強的正相關,也與氣道平滑肌層厚度呈現強的正相關。抗胸腺基質淋巴生成素抗體可明顯減少肺部氣道結構平滑肌層厚度,降低支氣管肺泡灌洗液中的基質金屬蛋白酶-2、基質金屬蛋白酶-9和肺部的結締組織生長因子、轉化生長因子-β。我們的總結中,胸腺基質淋巴生成素和胸腺基質淋巴生成素接受器可能是氣喘重要的生物指標。結締組織生長因子與基質金屬蛋白酶-9相關並參與氣喘氣道重塑的發病機制。抗胸腺基質淋巴生成素抗體或許對氣喘的氣道重塑具有預防作用,同時會影響基質金屬蛋白酶,結締組織生長因子、轉化生長因子-β等發炎性物質並且減少氣道結構改變。 | zh_TW |
dc.description.abstract | Asthma is a common inflammatory respiratory disease in children. Thymic stromal lymphopoietin (TSLP) is a cytokine interacting with thymic stromal lymphopoietin receptor (TSLPR) that plays an important role in pathogenesis in patients with asthma. However, the role of TSLP, TSLPR as the biomarkers in asthma remains unclear. The effects of anti-thymic stromal lymphopoietin antibody (anti-TSLP antibody) on MMP, CTGF and TGF-β in airway remodeling of asthma and the correlation between CTGF and MMP in airway remodeling of asthma were still unknown and worthy of investigation. In vitro study, we tried to investigate TSLP and TSLPR levels in asthmatic and non-asthmatic children. We drew blood samples to check TSLP, and measured expression of the TSLPR genes using reverse-transcription polymeric chain reaction (RT-PCR) and real time PCR. In vivo study, we established airway remodeling of asthma in murine model and investigated CTGF expression and its correlation to MMP and airway structural changes. We also examined preventive impact of ant-TSLP antibody on airway structural changes and determine influence of MMP, CTGF and TGF-β in airway remodeling of asthma. The results in vitro study, TSLP and TSLPR were significantly higher in asthmatic than non-asthmatic children. The results in vivo study in airway remodeling of asthma in murine model, CTGF expression significantly increased. CTGF expressions positively correlated with MMP-9, and thickness of smooth muscle layer. The impact of anti-TSLP antibody on airway remodeling, MMP-2, MMP-9 in BALF and lung CTGF, TGF-β expression significantly decreased. Airway structural changes of animals’ lungs show less thickness of smooth muscle layer. In our total conclusion, TSLP and TSLPR might be a significant disease biomarker for asthma. CTGF upregulation correlates with MMP-9 involved in pathogenesis of airway remodeling of asthma. Anti-TSLP antibody had preventive effect to decrease airway structural changes and inflammatory mediators of MMP, CTGF and TGF-β in airway remodeling of asthma. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T03:41:22Z (GMT). No. of bitstreams: 1 ntu-108-D01421014-1.pdf: 4274879 bytes, checksum: 6024b8e55248e726cd727faf39469ab8 (MD5) Previous issue date: 2019 | en |
dc.description.tableofcontents | Content………………………………………………………………........................1-3
中文摘要 ………………………………………………………...............................4-5 Abstract ………………………………………………………………......................6-7 Chapter 1 Introduction………………………………………………………………...8 1.1 Background……………………………………………………………….…..9 1.1.1 Pathogenesis of asthma……………………………………………9 1.1.2 Pathogenesis of airway remodeling in asthma…………….............9 1.1.3 The immune reaction of TSLP in asthma……..………………..…9 1.1.4 The importance of TSLP in asthma……………………………...11 1.2 Background of Pediatric asthma…………………………………………….12 1.2.1 Clinical evaluation and risk factor about TSLP in pediatric asthma……………………………………………………………12 1.2.2 Diagnosis, management and the importance of TSLP, TSLPR in pediatric asthma………………………………………………….13 Chapter 2 Aims….........................................................................................................17 2.1 Part 1………………………………………………………………………..18 2.1.1 TSLP and TSLPR might overexpress and influence immune modulation in the course of asthma in children………………….19 2.1.2 The purpose of part 1 study………………………………………21 2.2 Part 2……………………………………………………………………….21 2.2.1 Increased CTGF and MMP expression in a murine model of asthma might be associated with structural airway remodeling.....21 2.2.2 The purpose of part 2 study………………………………….…...23 2.3 Part 3………………………………………….…………………………….23 2.3.1 Ant-TSLP antibody may have effect on the CTGF, TGF-β and MMP that involved in airway remodeling of asthma...…………..23 2.3.2 The purpose of part 3 study……………………………………....26 Chapter 3 Methods and Materials…………………………………………………..27 3.1 Part1: Research of TSLP and TSLPR on asthmatic children in vitro study………………………………………………………...28 3.1.1 Participants and data collection…………………………………..28 3.1.2 Asthma Symptom Score………………………………………….29 3.1.3 Subgroup classification…………………………………………..29 3.1.4 Total IgE and Mite-specific IgE………………………………….30 3.1.5 Cytokine ELISA of TSLP……………………………………..…30 3.1.6 Molecular methods……………………………………………….31 3.1.6.1 PBMC isolation……………………………………………..31 3.1.6.2 RNA extraction……………………………………………...31 3.1.6.3 RNA product expression obtained from samples by Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR)………………………………………………..32 3.1.6.4 Analysis of TSLPR gene expression by Real-Time Polymerase Chain Reaction (Q-PCR)………33 3.1.7 Statistics………………………………………………………34 3.2 Part2: Established airway remodeling of asthma in murine model………....35 3.2.1 Animal models…………………………………………………...35 3.2.2 Airway responsiveness measurement……………………………36 3.2.3 Serum OVA-specific IgE………………………………………...36 3.2.4 Evaluation of bronchoalveolar lavage fluid (BALF)…………….37 3.2.5 Morphometric analysis…………………………………………..37 3.2.6 Western blot analysis…………………………………………….38 3.2.7 Statistical analysis………………………………………………..38 3.3 Part 3: Study on anti-TSLP antibody in airway remodeling of asthmatic mice in vivo study……………………………………………….......39 3.3.1 Murine model in airway remodeling of asthma…………….……39 3.3.2 Measurement of airway responsiveness by plethysmography after anti-TSLP antibody………………………………………....41 3.3.3 Evaluation of serum OVA-specific IgE after anti-TSLP antibody………………………………………………………….42 3.3.4 Evaluation of bronchoalveolar lavage fluid (BALF) after anti-TSLP antibody…..................................................................42 3.3.5 Evaulation of morphometric and IHC analysis after anti-TSLP antibody……………………………………………………...…43 3.3.6 Evaluation of western blot analysis after anti-TSLP antibody…..44 3.3.7 Statistical analysis………………………………………………..45 Chapter 4 Results…………………………………………………………………….46 4.1 Part 1: Results of TSLP and TSLPR on asthmatic children in vitro study….47 4.1.1 The demographic of participants………………………………...47 4.1.2 Analysis of total serum IgE and serum TSLP levels…………….48 4.1.3 RNA product expression obtained from samples………………..48 4.1.4 Analysis of TSLPR mRNA levels………………………..……...48 4.1.5 The clinical severity in asthmatic children…………………...….50 4.1.6 Strong correlation between TSLPR and IL-7Rα…………..…......50 4.1.7 Weak correlation between TSLPR and total IgE and between TSLPR and specific-IgE against Dermatophagoides pteronyssinus………………………………………….………..50 4.1.8 No correlation between TSLPR and TSLP………………………51 4.2 Part 2: Results about establishing airway remodeling of asthma in murine model……………………………………………………………….51 4.2.1 Airway responsiveness (Penh)………………………………….51 4.2.2 OVA-specific IgE………………………………………………51 4.2.3 Inflammatory cell infiltrate in BALF…………………………...51 4.2.4 Lung histology and morphometric analysis…………………….52 4.2.5 Western blot Analysis of CTGF………………………………..52 4.2.6 Concentrations of MMP-2 and MMP-9 in BALF……………...53 4.2.7 Correlation between CTGF, MMP and airway structural changes………………………………………………………….53 4.3 Part 3: Results of anti-TSLP antibody in airway remodeling of asthmatic mice in vivo study………………...………………………………....53 4.3.1 Airway responsiveness (Penh) after anti-TSLP antibody............53 4.3.2 Serum OVA-specific IgE after anti-TSLP antibody……………54 4.3.3 Differential cell count in BALF after anti-TSLP antibody...........54 4.3.4 Concentrations of MMP-2 and MMP-9 in BALF after anti-TSLP antibody…………………..........................................55 4.3.5 Lung histology, morphometric and ICH analysis after anti-TSLP antibody………………………………………………………...55 4.3.6 Western blot analysis of CTGF, TGF-β, TSLP after anti-TSLP antibody………………..……………………………………….56 Chapter 5 Discussion…………………………………………………………………58 Chapter 6 Prospective……………………………………………………………...…75 Chapetr 7 Reference……………………………………………………………..81-107 Chapter 8 Table………………………………………..……………………… 108-112 Chapter 9 Figure………………………………………………………………..113-129 Chapter 10入學後論文………………………………………………………..130-132 | |
dc.language.iso | en | |
dc.title | 研究胸腺基質淋巴生成素和胸腺基質淋巴生成素接受器在氣喘相關的調控與致病機轉 | zh_TW |
dc.title | Study on the regulatory role and pathogenesis of thymic stromal lymphopoietin and thymic stromal lymphopoietin receptor in asthma | en |
dc.type | Thesis | |
dc.date.schoolyear | 107-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 黃凱文,陳中明,莊雅惠,葉怡玲 | |
dc.subject.keyword | 氣喘,胸腺基質淋巴生成素,胸腺基質淋巴生成素接受器,氣道重塑,結締組織生長因子,基質金屬蛋白?,轉化生長因子-β, | zh_TW |
dc.subject.keyword | asthma,thymic stromal lymphopoietin,thymic stromal lymphopoietin receptor,airway remodeling,connective tissue growth factor,matrix metalloproteinase,transforming growth factor-β, | en |
dc.relation.page | 132 | |
dc.identifier.doi | 10.6342/NTU201900758 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2019-06-27 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 臨床醫學研究所 | zh_TW |
顯示於系所單位: | 臨床醫學研究所 |
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