新穎性長鏈烴基醯胺衍生物 WMJ-S-001 的抗發炎作用機轉探討

Anti-inflammatory mechanism of WMJ-S-001, a novel aliphatic hydroxamate derivative

陳威銓(Wei-Chung Chen)

指導教授 ： 許銘仁

臺北醫學大學/醫學院/醫學科學研究所/碩士(2017年)

摘要

近來，烴基醯胺 (hydroxamate)衍生物所含有的廣泛的藥理學性質在治療上，引起了相當大的關注。最近的研究指出它們在治療心血管疾病，關節炎和傳染病方面的發展潛能。然而，烴基醯胺衍生物的抗發炎作用的機制仍未清楚，因此為了開發新藥物以抑制異常活化的巨噬細胞，我們研究了一種新型長鏈烴基醯胺衍生物WMJ-S-001 (C31H44NO7)，並探討其抗發炎機制。本論文以LPS刺激RAW264.7小鼠巨噬細胞的發炎細胞模式，來探討WMJ-S-001對LPS所誘導COX-2表現與訊息傳遞的作用機轉。結果指出，WMJ-S-001會抑制LPS誘導的COX-2表現，其下游的p38MAPK、NF-κB次單位；p65和CCAAT/增強子結合蛋白 (C/EBPβ)磷酸化程度增加。而處理以p38MAPK抑製劑(p38MAPK抑製劑III)也能觀察其能明顯抑制LPS誘導的p65、C/EBPβ磷酸化和COX-2表現。同時WMJ-S-001也能抑制LPS誘導p65和C/EBPβ與COX-2啟動子的結合能力。而細胞轉染MAPK磷酸水解酶-1 (MKP-1 DN)突變質體後觀察到，WMJ-S-001抑制LPS誘導p38MAPK、CEBP、p65的磷酸化作用以及COX-2表現會被MKP-1-DN所回復。另一方面，WMJ-S-001也會誘導RAW264.7巨噬細胞中MKP-1活性的增加。綜合結果來說，WMJ-S-001可透過活化MKP-1，使p38MAPK去磷酸化，導致p65和C/EBPβ與COX-2啟動子的結合能力下降，進而抑制COX-2蛋白表現。因此，WMJ-S-001可能是減輕LPS相關炎症性疾病的潛在候選藥物。

關鍵字

烴基醯胺 ； 抗發炎作用

並列摘要

Hydroxamate derivatives have attracted considerable attention because of their broad pharmacological properties. Recent studies reported their potential use in the treatment of cardiovascular diseases, arthritis and infectious diseases. However, the mechanisms of the anti-inflammatory effects of hydroxamate derivatives remain to be elucidated. In an effort to develop a novel pharmacological agent that could suppress abnormally activated macrophages, we investigated a novel aliphatic hydroxamate derivative, WMJ-S-001 (C31H44NO7), and explored its anti-inflammatory mechanisms. RAW264.7 macrophages were exposed to LPS in the absence or presence of WMJ-S-001. COX-2 expression and signaling molecules activated by LPS were assessed. LPS-induced COX-2 expression was suppressed by WMJ-S-001. WMJ-S-001 inhibited p38MAPK, NF-κB subunit p65 and C/EBPβ phosphorylation in cells exposed to LPS. Treatment of cells with a p38MAPK inhibitor (p38MAPK inhibitor III) markedly inhibited LPS-induced p65 and C/EBPβ phosphorylation and COX-2 expression. LPS-increased p65 and C/EBPβ binding to the COX-2 promoter region was suppressed in the presence of WMJ-S-001. In addition, WMJ-S-001 suppression of p38MAPK, p65 and C/EBPβ phosphorylation, and subsequent COX-2 expression were restored in cells transfected with a dominant-negative (DN) mutant of MAPK phosphatase-1 (MKP-1). WMJ-S-001 also caused an increase in MKP-1 activity in RAW264.7 macrophages. WMJ-S-001 may activate MKP-1, which then dephosphorylates p38MAPK, resulting in a decrease in p65 and C/EBPβ binding to the COX-2 promoter region and COX-2 down-regulation in LPS-stimulated RAW264.7 macrophages. The present study suggests that WMJ-S-001 may be a potential drug candidate for alleviating LPS-associated inflammatory diseases.

並列關鍵字

hydroxamate ； COX-2

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