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標題: | 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 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 |
作者: | Hsiu-Pin Hung 洪琇品 |
指導教授: | 鄭景暉 |
關鍵字: | 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, 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), |
出版年 : | 2014 |
學位: | 碩士 |
摘要: | 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。 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. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58543 |
全文授權: | 有償授權 |
顯示於系所單位: | 臨床牙醫學研究所 |
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