前列腺素E2對人類牙髓細胞的調節作用:EP receptor被活化後的角色扮演及訊息傳導

Regulation of PGE2 on the Human Dental Pulp Cells : Role of EP Receptors Activation and Their Signaling

王彥臻(Yen-Chen Wang)

指導教授 ： 鄭景暉

國立臺灣大學/牙醫專業學院/臨床牙醫學研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.01389

摘要

實驗目的: 在牙髓受感染或發炎的過程中，前列腺素E2是重要因子來調節牙髓細胞的生理病理功能。本篇論文目的在探討前列腺素E2對人類牙髓細胞的調節作用、相關EP receptor被活化後的角色扮演及訊息傳導。
實驗方法: 本研究以前列腺素E2以及不同前列腺素E受體的促效劑(agonist)或是預先加入AC/PKA抑制劑三十分鐘後再給予前列腺素E2，來刺激培養的人類牙髓細胞，並觀察其細胞反應暨其訊息傳導路徑。
實驗結果:
1.前列腺素E2在我們所分析的濃度對牙髓細胞來說是抑制作用，且濃度越高抑制作用越明顯，此現象在cell migration及collagen turnover assay和鹼性磷酸酶染色都能觀察到。
2.加入EP2 agonist 19R-OH PGE2能影響人類牙髓細胞膠原蛋白之代謝，cell migration能力，以及鹼性磷酸酶的表現。
3.分別給予PKA抑制劑H89(濃度1μM)以及腺苷酸環化酶ACs抑制劑SQ22536似乎能部分逆轉原先前列腺素E2抑制cell migration的情形及抑制鹼性磷酸酶的表現情形。
4.AP-1 轉錄因子表現上升與改變，證實AP-1在前列腺素E2調控發炎反應扮演某種角色。
結論: 不論在ALP activity或者是cell migration的結果都可以指出前列腺素E2調節人類牙髓細胞這些作用主要以EP2受器為主，可能是藉由AC/PKA的路徑來調控。AP-1 轉錄因子表現上升改變，證實AP-1在前列腺素E2調控發炎反應扮演相當之角色。

關鍵字

牙髓細胞 ； PGE2 ； 前列腺素受體 ； 膠原蛋白 ； 鹼性磷酸酶AP-1 轉錄因子 ； 訊息傳導

並列摘要

Aim: Bacterial infection and mechanical injuries can cause pulp inflammation. Prostanoids production of pulp cells were involved in the inflammatory and healing processes of dental pulp. These contradictory actions of PGE2 further support the presence of diverse PGE2 receptor isoforms in different tissues or cells. However, the significance of EP receptors in human dental pulp has not yet been fully described. The aim of the study is to investigate the role of EP receptors and their signaling in regulation of the dental pulp.
Materials and Methods: Primary-cultured human dental pulp cells were treated with PGE2 or EP receptor agonist. In some experiments, cells were pretreated with H89 (a PKA inhibitor), or SQ22536 (an ACs inhibitor) 30 minutes before adding PGE2. Cell migration assay has been used to examine signaling events. Collagen content was determined by Sircol Collagen assay. Cell differentiation and mineralization was evaluated by alkaline phosphatase (ALP) staining. Changes in mRNA expression were determined by reverse-transcriptase polymerase chain reaction (RT-PCR).
Result:Cell under the treatment of PGE2 (0.1μM~10μM) showed a decrease of collagen formation and cell migration and also showed a decrease of ALP activity. CAY10598 (EP2 agonist) at low dose (0.5μM , 1μM) increased ALP activity. 19R-OH PGE2 (EP2 agonist) showed a decrease in cell migration assay and collagen formation assay . Pretreatment by H89 (a PKA inhibitor), or SQ22536 (an ACs inhibitor) were effective to reverse the inhibitory effect of PGE2 in ALP stain and in the cell migration assay. PGE2 can induce the expression of AP-1 transcription factors.
Conclusion: Signal transduction of PGE2 in human dental pulp cell is complex, AC/PKA may take part in this complex system via EP2. PGE2 may be involved in dental pulp inflammation and repair processes via induction of AP-1 transcription factors.

並列關鍵字

dental pulp cells ； PGE2 ； collagen ； alkaline phosphatase ； AP-1 transcription factors ； prostaglandin receptor

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