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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 張曉華 | zh_TW |
| dc.contributor.advisor | Hsiao-Hua Chang | en |
| dc.contributor.author | 洪歆雅 | zh_TW |
| dc.contributor.author | Hsin-Ya Hung | en |
| dc.date.accessioned | 2025-09-16T16:05:56Z | - |
| dc.date.available | 2025-09-17 | - |
| dc.date.copyright | 2025-09-16 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-07-24 | - |
| dc.identifier.citation | Amaravadi, R. K., & Winkler, J. D. (2012). Lys05: A new lysosomal autophagy inhibitor. Autophagy, 8(9), 1383–1384. https://doi.org/10.4161/auto.20958
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/99554 | - |
| dc.description.abstract | 4-甲基丙烯酰氧基偏苯三酸酐 (4-methacryloxyethyl trimellitic anhydride, 4-META) 是一種常見的功能性單體 (functional monomer)，被廣泛使用在牙科樹脂黏著劑 (resin adhesive) 中。然而，這些樹脂材料可能因鍵結不穩定、聚合不完全，或於口腔環境中長時間水解等因素，於填充牙齒中或是接觸牙齒表面時，經由牙本質小管滲入牙髓中，進而對牙髓細胞造成潛在的傷害。我們過去的研究結果顯示，4-META 會使人類牙髓細胞中活性氧生成量增加，氧化壓力上升，細胞週期調控失序，細胞存活率降低以及促發炎因子生成。然 而，4-META 對牙髓細胞的毒性及其促發炎反應機制尚未被釐清，細胞因應的調節防護作用也有待瞭解。細胞自噬 (autophagy) 被認為是細胞面對外在毒性或壓力刺激時的重要自我防 禦與存活的機制之一;細胞焦亡(pyroptosis) 是一種具高度調控性的細胞死亡方式，與促發炎反應密切相關。近年來，這兩種機制在多種疾病模型中均展現出調控細胞命運與組織損傷 的重要角色。因此，本研究旨在於探討細胞自噬及細胞焦亡對於 4-META 誘導之人類牙髓 細胞毒性及促發炎反應扮演之角色，我們期望進一步釐清4-META 之細胞毒性的作用機 轉，並為未來開發生物相容性更高的牙科修復材料提供依據及潛在應用方向 | zh_TW |
| dc.description.abstract | 4-Methacryloxyethyl trimellitic anhydride (4-META) is a widely used functional monomer in dental resin adhesives. However, these resin-based materials may release residual monomers due to unstable bonding, incomplete polymerization, or long-term hydrolysis under the moist environment of the oral cavity. These monomers can diffuse through dentinal tubules into the pulp tissue and potentially cause cytotoxic effects on dental pulp cells. Our previous studies have shown that 4- META increases intracellular reactive oxygen species (ROS), elevates oxidative stress, disrupts cell cycle regulation, decreases cell viability, and promotes the expression of pro-inflammatory cytokines in human dental pulp cells. However, the precise mechanisms underlying the cytotoxic and pro- inflammatory effects of 4-META remain unclear, and the potential cellular protective responses also warrant further investigation. Autophagy is regarded as an essential adaptive mechanism for cellular defense and survival under toxic or stress-inducing conditions. Pyroptosis, a highly regulated form of programmed cell death, is closely associated with inflammatory responses. In recent years, both autophagy and pyroptosis have been recognized as key regulators of cell fate and tissue injury in various disease models. Therefore, this study aims to investigate the roles of autophagy and pyroptosis in 4-META-induced cytotoxicity and inflammation in human dental pulp cells. We hope to elucidate the underlying mechanisms of 4-META toxicity and provide a scientific basis for developing dental restorative materials with improved biocompatibility. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-09-16T16:05:56Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-09-16T16:05:56Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員會審定書 i
中文摘要 ii Abstract iv 目次 vi 圖次 xi 表次 xiii 第一章 文獻回顧 1 1.1 序言 1 1.2 4-甲基丙烯酰氧基偏苯三酸酐 (4-methacryloxyethyl trimellitic anhydride, 4-META) 1 1.2.1 4-甲基丙烯酰氧基偏苯三酸酐介紹 1.2.2 4-META 的細胞毒性 1.3 細胞死亡 (Cell Death) 3 1.4 細胞自噬 (Autophagy) 4 1.5 細胞自噬重要相關分子 5 1.5.1 LC3-II 1.5.2 Beclin-1 1.5.3 p62 1.5.4 ATG5、ATG12 1.5.5 Cathepsin B 1.6 細胞焦亡 (Pyroptosis) 7 1.7 細胞焦亡重要相關分子 8 1.7.1 NOD-like receptor family pyrin domain-containing 3 (NLRP3) 1.7.2 NOD-like receptor family pyrin domain-containing 7 (NLRP7) 1.7.3 Absent in Melanoma 2 (AIM2) 1.7.4 caspase-1 1.7.5 Gasdermin D (GSDMD) 1.7.6 Interleukin-1β (IL-1β)、Interleukin-18 (IL18) 1.8 氧化壓力重要相關分子 11 1.8.1 Nuclear factor erythroid 2-related factor (Nrf2) 1.8.2 血鐵質氧化酶-1 (Heme oxygenase-1, HO-1) 1.9 發炎反應重要相關分子 12 1.9.1 環氧化酶 (Cyclooxygenase-2, COX-2) 1.9.2 Interleukin-6 (IL-6) 1.9.3 Interleukin-8 (IL-8) 第二章 實驗目的與假說 14 第三章 材料與方法 15 3.1 材料準備 15 3.1.1 樣本試劑 3.1.2 儀器設備 3.2 人類牙髓細胞 (human dental pulp cells, hDPCs) 培養 17 3.3 即時定量聚合酶連鎖反應 (Real-time quantitative PCR) 18 3.3.1 核糖核酸的萃取 (RNA extraction) 3.3.2 核糖核酸的定量 (RNA quantification) 3.3.3 反轉錄 (RNA transcription) 3.3.4 即時定量聚合酶連鎖反應 (Real-time quantitative PCR) 3.4 西方點墨法 (Western blot) 21 3.4.1 蛋白質萃取 (protein extraction) 3.4.2 蛋白質定量 (protein quantification) 3.4.3 十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 (sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE) 3.4.4 轉漬 (Transfer gel) 3.4.5 阻斷及抗體雜合 (Blocking and antibody hybridization) 3.4.6 化學冷光影像擷取 (Chemiluminescence photography) 3.5 免疫螢光分析法 (Immunofluorescence assay) 24 3.5.1 細胞培養 3.5.2 免疫螢光染色 (Immunofluorescence staining) 3.6 細胞存活率分析:MTT assay 25 3.7 紅色溶酶體追蹤染劑 (Lysotracker Red Staining) 26 3.7.1 細胞培養 3.7.2 工作溶液配置 3.7.3 紅色溶酶體追蹤染劑 (Lysotracker Red staining) 3.8 統計分析 27 第四章 實驗結果 28 4.1 4-META 濃度對於細胞自噬之影響 28 4.1.1 對於 LC3-II 基因及蛋白質表現之影響 4.1.2 對於 Beclin-1 基因及蛋白質表現之影響 4.1.3 對於 p62 基因及蛋白質表現之影響 4.1.4 對於 ATG5 基因及蛋白質表現之影響 4.1.5 對於 ATG12 基因及蛋白質表現之影響 4.1.6 對於 Cathepsin B 基因及蛋白質表現之影響 4.2 加入 3-methyladenine 後 4-META 濃度對於人類牙髓細胞的存活率之影響:MTT assay 29 4.3 加入 3-methyladenine 後 4-META 濃度對於人類牙髓細胞形態之影響 30 4.4 加入 Lys05 後 4-META 濃度對於人類牙髓細胞的存活率之影響:MTT assay 30 4.5 加入 Lys05 後 4-META 濃度對於人類牙髓細胞形態之影響 31 4.6 4-META 濃度對於溶酶體活性之影響 31 4.7 加入 Lys05 後 4-META 濃度對於溶酶體活性之影響 31 4.8 4-META 濃度對於細胞焦亡之影響 31 4.8.1 對於 NLRP3 基因及蛋白質表現之影響 4.8.2 對於 NLRP7 基因及蛋白質表現之影響 4.8.3 對於 AIM2 基因及蛋白質表現之影響 4.8.4 對於 caspase-1 基因及蛋白質表現之影響 4.8.5 對於 GSDMD 基因及蛋白質表現之影響 4.8.6 對於 IL-1β 基因及蛋白質表現之影響 4.8.7 對於 IL-18 基因及蛋白質表現之影響 4.9加入 MCC950 後 4-META 濃度對於人類牙髓細胞的存活率之影響: MTT assay 33 4.10 加入 MCC950 後 4-META 濃度對於人類牙髓細胞形態之影響 33 4.11 加入 parthenolide 後 4-META 濃度對於人類牙髓細胞的存活率之 影響:MTT assay 34 4.12 加入 parthenolide 後 4-META 濃度對於人類牙髓細胞形態之影響 34 4.13 加入 Lys05 後 4-META 濃度對於發炎作用及氧化壓力之影響 35 4.14 加入 parthenolide 後 4-META 濃度對於發炎作用之影響 36 第五章 討論 38 5.1 與細胞自噬之關係 38 5.2 與細胞焦亡之關係 40 5.3 與發炎作用之關係 41 5.4 與氧化壓力之關係 42 5.5 與溶酶體之關係 43 第六章 結論 45 參考文獻 46 表格 54 附圖 62 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 4-甲基丙烯酰氧基偏苯三酸酐 | zh_TW |
| dc.subject | 人類牙髓細胞 | zh_TW |
| dc.subject | 細胞毒性 | zh_TW |
| dc.subject | 細胞自噬 | zh_TW |
| dc.subject | 細胞焦亡 | zh_TW |
| dc.subject | 發炎反應 | zh_TW |
| dc.subject | 氧化壓力 | zh_TW |
| dc.subject | pyroptosis | en |
| dc.subject | human dental pulp cells | en |
| dc.subject | cytotoxicity | en |
| dc.subject | oxidative stress | en |
| dc.subject | inflammation | en |
| dc.subject | autophagy | en |
| dc.subject | 4-META | en |
| dc.title | 4-META對人類牙髓細胞的影響:自噬與焦亡作用的相關探討 | zh_TW |
| dc.title | Exploring the Effects of 4-META on Human Dental Pulp Cells: The Roles of Autophagy and Pyroptosis | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.coadvisor | 鄭景暉 | zh_TW |
| dc.contributor.coadvisor | Jiiang-Huei Jeng | en |
| dc.contributor.oralexamcommittee | 王彥雄;李苑玲;張美姬 | zh_TW |
| dc.contributor.oralexamcommittee | Yan-Hsiung Wang;Yuan-Ling Lee;Mei-Chi Chang | en |
| dc.subject.keyword | 4-甲基丙烯酰氧基偏苯三酸酐,人類牙髓細胞,細胞毒性,細胞自噬,細胞焦亡,發炎反應,氧化壓力, | zh_TW |
| dc.subject.keyword | 4-META,human dental pulp cells,cytotoxicity,autophagy,pyroptosis,inflammation,oxidative stress, | en |
| dc.relation.page | 118 | - |
| dc.identifier.doi | 10.6342/NTU202502127 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2025-07-24 | - |
| dc.contributor.author-college | 醫學院 | - |
| dc.contributor.author-dept | 臨床牙醫學研究所 | - |
| dc.date.embargo-lift | 2025-09-17 | - |
| Appears in Collections: | 臨床牙醫學研究所 | |
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| File | Size | Format | |
|---|---|---|---|
| ntu-113-2.pdf Access limited in NTU ip range | 6.55 MB | Adobe PDF |
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