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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 姚宗珍(Jane Chung-Chen Yao) | |
| dc.contributor.author | Liang-Jou Kuo | en |
| dc.contributor.author | 郭良柔 | zh_TW |
| dc.date.accessioned | 2021-06-07T17:32:49Z | - |
| dc.date.copyright | 2020-09-02 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-07-08 | |
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'Anti-M3 muscarinic cholinergic autoantibodies from patients with primary Sjögren's syndrome trigger production of matrix metalloproteinase-3 (MMP-3) and prostaglandin E2 (PGE2) from the submandibular glands.' archives of oral biology 56.5 (2011): 413-420. Roberts, W. E., D. C. Chase, and W. S. S. Jee. 'Counts of labelled mitoses in the orthodontically-stimulated periodontal ligament in the rat.' Archives of oral biology 19.8 (1974): 665-670. Roberts, W. Eugene, et al. 'Remodeling of mineralized tissues, part II: control and pathophysiology.' Seminars in orthodontics. Vol. 12. No. 4. WB Saunders, 2006. Sasaki, S., et al. 'Detection of stromelysin in synovial fluid and serum from patients with rheumatoid arthritis and osteoarthritis.' Clinical rheumatology 13.2 (1994): 228-233. Sakamoto, Seizaburo, and Masako Sakamoto. 'Biochemical and immunohistochemical studies on collagenase in resorbing bone in tissue culture: A novel hypothesis for the mechanism of bone resorption.' 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'TIMP-3 and MMP-3 contribute to delayed inflammation and hippocampal neuronal death following global ischemia.' Experimental neurology 216.1 (2009): 122-131. Woo, Moon‐Sook, et al. 'Inhibition of MMP‐3 or‐9 suppresses lipopolysaccharide‐induced expression of proinflammatory cytokines and iNOS in microglia.' Journal of neurochemistry106.2 (2008): 770-780. Ren, Yijin, et al. 'Optimum force magnitude for orthodontic tooth movement: a mathematic model.' American journal of orthodontics and dentofacial orthopedics 125.1 (2004): 71-77. Sugiyama, Yuriko, et al. 'The level of cathepsin B in gingival crevicular fluid during human orthodontic tooth movement.' The European Journal of Orthodontics 25.1 (2003): 71-76. Zhu, Chaoyong, et al. 'Allele-specific MMP-3 transcription under in vivo conditions.' Biochemical and biophysical research communications 348.3 (2006): 1150-1156. Zhang, Shiqian, et al. 'Osteoclast regulation of osteoblasts via RANK RANKL reverse signal transduction in vitro.' Molecular medicine reports 16.4 (2017): 3994-4000. 翁竹音(2005). 週期性壓力刺激對類成骨母細胞基因表現的影響. 廖志清(2009).探討機械力刺激骨細胞之基質金屬蛋白酵素-3調控機制 蔡淑珺(2010). 週期性壓力刺激對於細胞外基質金屬蛋白酶-3 基因 啟動子在老鼠類骨母細胞內的調控 蕭琁憶(2012). 基質金屬蛋白酶 3 基因啟動子- 綠螢光蛋白之基因轉殖小鼠經矯正施力後之螢光表現 蔡孟桓(2013).以大鼠動物測試低能量雷射治療矯正牙齒移動及固持之效用 林昇進(2014). 基因金屬蛋白酶-3基因啟動子-綠螢光蛋白』 之基因轉殖小鼠經矯正施力後之螢光表現 廖珮雯(2015). 基質金屬蛋白酶-3 基因啟動子- 基因轉殖小鼠及基因轉 殖大鼠對矯正機械性刺激之表現 陳泱錚(2017). 低能量雷射治療對大鼠矯正牙齒移動模式之影響 游元亨(2019). 藉由MMP-3-GFP基因轉殖小鼠及MMP-3-Luc-GFP大鼠探討MMP-3在矯正性牙齒移動的表現 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/15315 | - |
| dc.description.abstract | MMP-3為一種內切蛋白酶。在不同層面上調控此分子的表現和活性也影響骨質塑形/重塑。在一般生理環境下可以看到此分子表現外，在發炎環境下，例如類風濕性關節炎中，也會因疾病的進展，關節腔或血液內之MMP-3濃度增加。而在特定的機械壓力或是化學刺激之不同實驗模型中，24小時內都可以見到MMP-3明顯的變化。 本實驗使用Mmp-3-luciferase-GFP基因轉殖大鼠，利Luciferase可由冷光偵測觀察MMP-3在受力刺激後時間與空間的相應關係。細胞實驗中也使用關節滑液腔取出培養之纖維母細胞初代細胞，以不同量化報導基因蛋白質的方法，找出此報導基因蛋白質的半生期，以期可以呼應動物實驗中的時間、受力大小、空間分佈等參數之相關性。動物實驗結果發現在十五天的矯正力量施予期間，Luciferase冷光訊號於施力後第一天訊號最為明顯，並於第三天回到與原始基礎訊號相同;即使期間每三天給予低能量雷射照射或是重新給予矯正施力，皆無法促使報導基因再度活化、冷光訊號強度上升。在翻開實驗大鼠臉頰觀察冷光訊號表現時，也可以見到在施予矯正力量後的一天，Luciferase冷光訊號確實出現於受力的第一大臼齒上，並且訊號強度於第一天時強度最強，在放掉力量後三小時開始下降，直至八小時回到與原始基礎訊號相同。而細胞實驗中，給 LPS濃度達 1 ng/ml，刺激八小時後在冷光強度訊號(luciferase assay)可以見到上升趨勢。在西方墨點法中，當LPS濃度為0.5 ng/ml與1 ng/ml，刺激八小時，可以偵測到較為明顯之MMP-3蛋白質表現。 在本實驗動物模型中，可以看到在牙齒受到持續性矯正力或是間歇性矯正力刺激時， MMP-3皆在初期力量刺激時即表現，一天可達高峰，移除力量後很快消失。 | zh_TW |
| dc.description.abstract | MMP-3 is an endoproteinase which the activity is regulated at different levels to influence bone modeling and remodeling. In inflammation, such as rheumatoid arthritis, the increased level of MMP-3 correlated with the severity of diseases. With specific mechanical force or chemical stimulation in different models, MMP-3 significantly and quickly increased within 24 hours. By observing luciferase driven by Mmp-3-promoter in a transgenic rat model, we studied the temporospacial expression of MMP-3 during orthodontic force stimulation. In vivo, luciferase increased signficantly after 1 day of orthodontic force stimulation then dropped back to baseline on the 3rd day of a fifteen day experiment. Neither low level laser applied every three days, nor orthodontic force reactivation did enhance the signal. Location of luminance was at the molar under orthodontic force for protraction, when cheeks were reflected for a direct view of molars. Once the force removed, the signal decreased and dropped to baseline level after 8 hours. In vitro, isolated synovial fibroblast from transgenic rat was used to measure the rate of protein degradation up on LPS stimulation. A trend of bioluminescence signal was detected after stimulation of LPS 1 ng/ml at 8 hours. In Western blot assay, obvious MMP-3 protein band was shown when stimulation by LPS 1 ng/ml and 0.5 ng/ml at 8 hours. In our study, we demonstrate that MMP-3 was expressed at the initial phase of mechanical stimulation. Either the continuous force or interrupted force, the bioluminescence signal of luciferase was enhanced after 1 day force application. This rise was quickly tuned down after 3 hours of force termination. Therefore, the regulation of this MMP-3 expression is tightly controlled in a timely fashion. Details of this regulatory mechanism warrant further studies. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-07T17:32:49Z (GMT). No. of bitstreams: 1 U0001-0507202021475300.pdf: 7176870 bytes, checksum: 2a8fddae3026d1e3c7234337dc432900 (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 口試委員審定書 I 中文摘要 II ABSTRACT III 目 錄 V 圖 目 錄 VII 表 目 錄 VIII 第一章 引言 1 1.1 矯正力量造成之牙齒移動之周遭環境與基本原理 1 1.1.1牙齒支持組織(牙周韌帶與齒槽骨)的組成 1 1.2基質金屬蛋白酶家族 2 1.2.1 MMP-3(stromelysin-1) 3 1.2.2剔除MMP-3之相關研究 5 1.3基質金屬蛋白酶 MMP-3與關節炎 5 1.4不同實驗模型引發MMP-3的表現-機械力刺激與化學刺激 6 1.4.1 MMP-3的調控機制 7 第二章 實驗目的 9 第三章 研究方法 10 3.1動物實驗 10 3.1.1基因轉殖大鼠之麻醉與口內裝置設置 10 3.1.2 MMP-3之偵測與量化 10 3.1.3 矯正牙齒移動(orthodontic tooth movement, OTM)之模型 11 3.1.3.1雷射對矯正牙齒移動之影響 11 3.1.3.2不同IVIS視野下對於影像判讀的差異 12 3.1.3.3矯正力量給予時間對MMP-3分子表現之影響 12 3.1.3.4彈簧施力與否對MMP-3分子表現之影響 12 3.2細胞實驗 13 3.2.1實驗大鼠關節腔滑液纖維母細胞之獲取 13 3.2.2關節腔滑液纖維母細胞初代細胞之培養 13 3.2.3量化報導基因蛋白質 13 3.2.3.1蛋白質量化-西方墨點法 14 3.2.3.2蛋白質量化-Luciferase assay 15 第四章 研究結果 16 4.1動物實驗 16 4.1.1雷射對矯正牙齒移動之影響 16 4.1.2不同IVIS視野下對於影像判讀的差異 17 4.1.3矯正力量給予時間對MMP-3分子表現之影響 17 4.1.4彈簧施力與否對MMP-3分子表現之影響 18 4.2細胞實驗 21 4.2.1關節腔滑液纖維母細胞初代細胞之培養 21 4.2.2蛋白質量化-西方墨點法 21 4.2.3蛋白質量化-Luciferase assay 22 第五章 討論 23 5.1 MMP-3與矯正牙齒移動(OTM) 23 5.2雷射對矯正牙齒移動之影響 23 5.3矯正力量給予時間對MMP-3分子表現之影響 24 5.4彈簧施力與否對MMP-3分子表現之影響 26 5.5細胞實驗 31 5.5.1蛋白質量化-西方墨點法與Luciferase assay 32 第六章 結論 34 6.1 動物實驗結論 34 6.2 細胞實驗結論 34 第七章 未來研究方向 35 參考文獻 36 附 錄 44 圖 一：大鼠口內裝置-SPLIT MOUTH DESIGN 44 圖 二：MMP-3轉殖基因序列示意圖 45 圖 三 : LUCIFERIN 引發 LUCIFERASE / MMP-3表現及其偵測 45 圖 四 : IVIS活體影像系統偵測之初始影像 45 圖 五 : IVIS活體影像系統視野示意圖 46 圖 六 : IVIS活體影像系統偵測所釋出之初步冷光影像圖 46 圖 七：實驗大鼠NO.78 OTM+LLLT連續十五天的IVIS影像 *WILD TYPE:此為NO.59實驗大鼠。此大鼠未經基因轉殖， 故可以見到其並未有IVIS冷光訊號。 47 圖 八 : 實驗大鼠NO.78 OTM+LLLT連續十五天冷光訊號量化圖 49 圖 九：IVIS活體影像系統不同視野下的照射情形 50 圖 十：IVIS SPECTRUM軟體重疊影像 51 圖 十一：矯正力量給予時間對MMP-3分子表現實驗之IVIS活體影像 52 圖 十二：矯正力量給予時間對MMP-3分子表現實驗之光子數量圖。 56 圖 十三：矯正力量給予時間對MMP-3分子表現實驗，翻開臉頰與否之光子數量比較。 57 圖 十四：彈簧施力與否對MMP-3分子表現之十五天影像變化 58 圖 十五：彈簧施力與否對MMP-3分子表現之十五天光子數量圖 60 圖 十六 : 彈簧施力與否對MMP-3分子表現，左側彈簧ACTIVATION與否之光子數量比較圖(N=2) 61 圖 十七 : 彈簧施力與否對MMP-3分子表現，實驗大鼠對照側與實驗側的光子數量比較圖。(N=2) 63 圖 十八 : 彈簧施力與否對MMP-3分子表現，左側彈簧ACTIVATION與否之光子數量比較圖(N=3) 65 圖 十九 : 延續加入圖 十八實驗大鼠右側的光子數量來做討論。 67 圖 二十: 初代細胞培養之結果 77 圖 二十一: 西方墨點法 78 表 一 : 雷射對矯正牙齒移動之影響實驗流程表 79 表 二 : 矯正力量給予時間對MMP-3分子表現之影響實驗流程表 80 表 三 : 彈簧施力與否對MMP-3分子表現之影響實驗流程表 80 表 四：西方墨點法使用之溶液 81 表 五：西方墨點法使用之一級抗體與二級抗體 83 表 六: LUCIFERASE ASSAY 84 | |
| dc.language.iso | zh-TW | |
| dc.title | 以MMP-3啟動-Luc-GFP報導基因於轉殖大鼠及其初代細胞研究受刺激後表達的時間特性 | zh_TW |
| dc.title | Temporal Expression of MMP-3 Promotor-Reporter under Stimulation -in Vivo and in Vitro | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.advisor-orcid | 姚宗珍(0000-0002-2293-7000) | |
| dc.contributor.oralexamcommittee | 王詩凱(Shih-Kai Wang),宋向軒(Hsiang-Hsuan Sung) | |
| dc.subject.keyword | 基質金屬蛋白酶,基因轉殖大鼠,牙齒矯正,關節滑液纖維母細胞,冷光素酶, | zh_TW |
| dc.subject.keyword | matrix metalloproteinase,transgenic rat,orthodontic force,synovial fibroblast,luciferase, | en |
| dc.relation.page | 84 | |
| dc.identifier.doi | 10.6342/NTU202001328 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2020-07-08 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
| 顯示於系所單位： | 臨床牙醫學研究所 | |
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| U0001-0507202021475300.pdf 目前未授權公開取用 | 7.01 MB | Adobe PDF |
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