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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 鄭景暉 | |
dc.contributor.author | Hsiu-Pin Hung | en |
dc.contributor.author | 洪琇品 | zh_TW |
dc.date.accessioned | 2021-06-16T08:19:14Z | - |
dc.date.available | 2014-02-25 | |
dc.date.copyright | 2014-02-25 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-02-10 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58543 | - |
dc.description.abstract | 實驗目的: 在牙髓中,介白素-1β(IL-1β)是一種重要的發炎介質。 IL-1β會刺激環氧酶-2(COX-2)並產生前列腺素 (prostanoids),進而影響牙髓的發炎和癒合過程。然而,當人類的牙髓組織發炎時, IL-1β對於細胞間黏附分子-1(ICAM-1)和單核細胞趨化蛋白-1 (MCP-1)的影響以及其與PI3K/Akt (也叫做Protein Kinase B)和MEK/ERK 1/2訊息傳遞路徑和COX活化之關係仍不完全清楚。
實驗方法: 在不同的時間點使用aspirin (COX抑制劑)或是LY294002 (PI3k/Akt抑制劑)或是U0126 (MEK/ERK 1/2抑制劑)加在人類的牙髓細胞中,檢查IL-1β是否經由這幾種的訊息傳遞路徑來調控牙髓細胞ICAM-1與MCP-1。用MTT的方法檢查是否這些IL-1β和這些抑制劑是否有細胞毒性。用反轉錄聚合酶連鎖反應的方法(RT-PCR)和西方點墨法(western blot)檢查IL-1β是否有刺激牙髓細胞ICAM-1和MCP-1的基因表現和蛋白質產生。用磷酸化的Akt和ERK 1/2抗體以西方點墨法確認IL-1β是否可活化這幾種的訊息傳遞路徑。 實驗結果: IL-1β會刺激牙髓細胞ICAM-1和MCP-1的基因表現和蛋白質產生。Aspirin同時處理會讓IL-1β刺激之牙髓細胞ICAM-1和MCP-1的基因表現增加。而這些訊息傳遞路徑有經過PI3K/Akt和MEK/ERK 1/2。另外, LY294002和U0126都可抑制IL-1β刺激之牙髓細胞ICAM-1和MCP-1的基因表現增加。 實驗結論: 這些結果表示,IL-1β在牙髓發炎和修復的過程中會產生ICAM-1,MCP-1的基因表達與蛋白質產生。這些過程都和PI3K/Akt和MEK/ERK 1/2的訊息傳遞訊息和COX的活化有關。這些結果對於牙髓發炎的治療都是有幫助的。 | zh_TW |
dc.description.abstract | Aim: Interleukin-1β (IL-1β) is an important inflammatory mediator of the dental pulp. IL-1β may stimulate cyclooxygenase-2 (COX-2) and prostanoids production of pulp cells and affect the inflammatory and healing processes of dental pulp. However, the effects of IL-1β on Intercellular Adhesion Molecule-1 (ICAM-1) and Monocyte Chemotactic Factor-1 (MCP-1) of human dental pulp cells and its relation to PI3K/Akt and MEK/ERK 1/2 signaling and COX activation are still not clear.
Materials and Methods: Human dental pulp cells were treated with IL-1β with/without pretreatment and co-incubation by aspirin (a COX inhibitor) or LY294002 (a PI3K/Akt inhibitor) or U0126 ( a MEK/ERK 1/2 inhibitor) for different time periods. Viable cell number was evaluated by 3- (4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The expression of ICAM-1 and MCP-1 mRNA was tested by reverse transcriptase – polymerase chain reaction (RT-PCR). ICAM-1 and MCP-1 protein expression was examined by western blotting. Phosphorylation of Akt and ERK 1/2 was measured by western blot. Results: IL-1β showed little cytotoxicity to pulp cells. It stimulated ICAM-1 and MCP-1 mRNA and protein expression. Aspirin enhanced the IL-1β-induced ICAM-1 and MCP-1 mRNA expression and protein production. IL-1β rapidly activated Akt and ERK 1/2. Both LY294002 and U0126 attenuated the IL-1β-induced ICAM-1and MCP-1 mRNA expression. Conclusions: These results reveal that IL-1β may involve in the pulpal inflammatory and healing processes by stimulation of ICAM-1and MCP-1 mRNA expression and protein production. These events are associated with differential activation of PI3K/Akt and MEK/ERK 1/2 signaling and COX. These results are useful for future treatment of pulpal inflammatory diseases. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T08:19:14Z (GMT). No. of bitstreams: 1 ntu-103-R00422019-1.pdf: 3232912 bytes, checksum: ed7f21c03b34666195a565139752fee6 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 口試委員審定書………………………………………………………i
誌謝……………………………………………………………………ii 中文摘要………………………………………………………………iii Abstract………………………………………………………………iv 第一章 前言………………….………………………………………1 第二章 文獻回顧…………….………………………………………2 2.1 牙髓的發炎反應(Inflammatory response of the dental pulp)……2 2.2 介白素-1β (Interleukin-1β) (IL-1β) 在發炎反應的角色……2 2.3 介白素-1 (Interleukin-1) (IL-1)………………………………3 2.4 介白素-1之生理功能………………………………………………5 2.5 細胞間黏附因子-1 (intercellular adhesion molecule-1) (ICAM-1)……………6 2.6 單核細胞趨化蛋白-1 (Monocyte Chemotactic Factor-1) (MCP-1)…………6 2.7 Protein kinase B (PKB) …………………………………………7 2.8 Mitogen-activated protein kinase (MAPK) pathway…………7 2.9 牙髓內IL-1β的訊息傳遞路徑………………………………………8 第三章 研究假說和實驗目的……………………………………………9 第四章 材料和方法………………………………………………………10 4.1 藥劑Reagent…………………………………………………………10 4.2培養人類牙髓細胞……………………………………………………10 4.3藥物對人類牙髓細胞影響(cell morphology)………………………11 4.4 藥物的細胞毒性(MTT assay)………………………………………11 4.5反轉錄聚合酶連鎖反應(Reverse Transcription Polymerase Chain Reaction) (RT-PCR)...11 4.6西方點墨法(Western Blot)…………………………………………14 4.7統計分析………………………………………………………………17 第五章 結果………………………………………………………………18 5.1人類牙髓細胞的型態觀察……………………………………………18 5.2人類牙髓細胞存活率檢測 (MTT assay)……………………………18 5.3反轉錄聚合酶連鎖反應 (RT-PCR)…………………………………18 5.4西方點墨法 (western blot)………………………………………19 第六章 討論……………………………………………………………21 6.1 牙髓中的ICAM-1…………………………………………………21 6.2 牙髓中的MCP-1……………………………………………………21 6.3 牙髓中的IL-1β與ICAM-1 , MCP-1和COX-2的活化……………22 6.4 牙髓中的IL-1β與ICAM-1 , MCP-1和PI3K/ Akt的活化………23 6.5 牙髓中的IL-1β與 ICAM-1 , MCP-1和MEK/ ERK 1/2的活化…23 第七章 結論……………………………………………………………25 References………………………………………………………27 圖 次 Cell morphology…………………………………………………………41 圖1 加入不同濃度的IL-1β,人類的牙髓細胞形態…………………………………………41 圖2 加入5ng/ml的IL-1β和100 & 200μM的aspirin,人類牙髓細胞形態…………42 圖3 加入5ng/ml的IL-1β和10 & 20μM的LY294002,人類牙髓細胞形態…………43 圖4 加入5ng/ml的IL-1β和10 &20μM的U0126,人類牙髓細胞形態………………44 MTT assay…………………………………………………………45 圖5 加入不同濃度的IL-1β,人類牙髓細胞存活率檢測…………………………………45 圖6加入5ng/ml的IL-1β和100 & 200μM的Aspirin,人類牙髓細胞存活率檢測…………46 圖7 加入5ng/ml的IL-1β和10 & 20μM的LY294002,人類牙髓細胞存活率檢測…………47 圖8 加入5ng/ml的IL-1β和10 & 20μM的U0126,人類牙髓細胞存活率檢測……………48 RT-PCR……………………………………………………………………….49 圖9 反轉錄聚合酶連鎖反應………………………………………………49 圖10 反轉錄聚合酶連鎖反應………………………………………………50 圖11 反轉錄聚合酶連鎖反應………………………………………………51 圖12 反轉錄聚合酶連鎖反應………………………………………………52 Western blot…………………………………………………………………53 圖13 西方點墨法………………………………………………………………53 圖13 西方點墨法………………………………………………………………54 | |
dc.language.iso | zh-TW | |
dc.title | 介白素-1β對人類牙髓細胞的細胞間黏附因子-1和單核細胞趨化蛋白-1的影響:PI3K/Akt和MEK/ERK 1/2訊息路徑與環氧酶的角色 | zh_TW |
dc.title | Effect of interleukin-1β on the ICAM-1 and MCP-1 expression of human dental pulp cells : Role of PI3K/Akt and MEK/ERK 1/2 Signaling
and Cyclooxygenase | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張美姬,林俊彬,黃富美,張育超 | |
dc.subject.keyword | PI3K/Akt,MEK/ERK 1/2,環氧?,牙髓細胞,發炎,介白素-1β,細胞間黏附分子-1,單核細胞趨化蛋白-1, | zh_TW |
dc.subject.keyword | PI3K/Akt,MEK/ERK 1/2,Cyclooxygenase (COX),Dental pulp cells,Inflammation,Interleukin-1β(IL-1β),Intercellular adhesion molecule-1 (ICAM-1),Monocyte Chemotactic Factor-1 (MCP-1), | en |
dc.relation.page | 54 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-02-10 | |
dc.contributor.author-college | 牙醫專業學院 | 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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