利多卡因誘導人類胚胎上顎間質細胞凋亡之研究

Lidocaine Induced Apoptosis in Human Embryonic Palatal Mesenchymal Cell

藍正邦(Cheng-Pang Lan)

指導教授 ： 郭彥彬

國立臺灣大學/牙醫專業學院/臨床牙醫學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.10711

摘要

牙科治療裡，利多卡因最常被用在牙科局部麻醉中。伴隨而來的併發症有顏面神經損傷、牙關緊閉、軟組織傷害等。文獻中探討利多卡因對細胞毒性與死亡的機轉並不十分清楚。
實驗目的：本研究探討以人類胚胎上顎間質細胞作為模式，了解利多卡因是否能造成口腔細胞凋亡或細胞自噬反應。同時更進一步研究其細胞凋亡背後的機轉與路徑。
材料與方法：利用細胞活性分析、西方點墨法、流式細胞分析TUNEL染色，不同濃度與時間，測試利多卡因對人類胚胎上顎間質細胞的影響
結果： 利多卡因會誘導人類胚胎上顎間質細胞進行細胞凋亡與自體吞噬反應，凋亡細胞與時間濃度增加成正比。不過，加入Caspase-8 抑制劑, Z-IETD-FMK, Caspase-9抑制劑, Z-LEHD-FMK, Pan-caspase抑制劑, Q-VD-OPH，都無法阻擋細胞進行細胞凋亡。此外加入ROS抑制劑與MAPK抑制劑也同樣無法使細胞活性恢復。另外，根據西方點墨法的結果，利多卡因同時也能誘導人類胚胎上顎間質細胞進行細胞自體吞噬反應。
結論：利多卡因會誘導人類胚胎上顎間質細胞進行細胞凋亡與細胞自體吞噬反應，但需要更多的研究來了解利多卡因對細胞死亡的機轉。

關鍵字

利多卡因 ； 人類胚胎上顎間質細胞 ； 細胞凋亡 ； 細胞自體吞噬反應

並列摘要

Lidocaine is one of the most used local anesthetics in dentistry. Although this is rare, a number of potential complications are associated with the administration of local anesthetics including facial nerve paralysis, trismus, soft-tissue injury, pain on injection.
Objectives：We investigated the mechanism of Lidocaine induced apoptosis in human embryonic palatal mesenchymal cells(HEPM) and try to identify possible signaling pathways that mediate their effects. In addition, we want to investigate any activation of autophagy in human embryonic palatal mesenchymal cells treated with Lidocaine.
Methods: Cell viability assay, flow cytometry, TdT-mediated dUTP nickend labeling assay and Western blot analysis were used to investigate the effects of Lidocaine on human embryonic palatal mesenchymal cells.
Results：We found Lidocaine induced apoptosis in dose and time dependent manner. However, after pretreated Caspase-8 inhibitor, Z-IETD-FMK, Caspase-9 inhibitor, Z-LEHD-FMK, Pan-caspase inhibitor, Q-VD-OPH then incubated in Lidocaine with HEPM, neither inhibitors can block Lidocaine induced apoptosis. In addition, ROS inhibitors and MAPK family inhibitors also failed to reverse cell viability in HEPM treated with Lidocaine. Meanwhile, western blotting showed that LC3-II was increased from the conversion of LC3-I in time dependent manners. It means Lidocaine also elicited autophagy in HEPM.
Conclusion：It is concluded that Lidocaine induced both apoptosis and autophagy in human embryonic palatal mesenchymal cells. Further research may need to elucidate the Lidocaine induced cell death.
Key words: Lidocaine, HEPM, Apoptosis, Autophagy

並列關鍵字

Lidocaine ； HEPM ； Apoptosis ； Autophagy

參考文獻

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