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系統識別號 U0026-0812200910450787
論文名稱(中文) 研究beraprost在HEL細胞中所活化的MAPK與PI3K訊息傳遞路徑
論文名稱(英文) Activation of both MAPK and PI3K signaling pathways by beraprost in human erythroleukemia cells
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 91
學期 2
出版年 92
研究生(中文) 呂湘敏
研究生(英文) Hsiang-Min Lu
學號 s2690408
學位類別 碩士
語文別 英文
論文頁數 79頁
口試委員 口試委員-呂增宏
指導教授-簡偉明
口試委員-簡伯武
中文關鍵字 肺性高血壓  花生四烯酸 
英文關鍵字 Prostacyclin  arachidonic acid  PKA  cAMP  IP receptor  beraprost  HEL cells  PI3K  MAPK  PKC 
學科別分類
中文摘要 Prostacyclin (PGI2) 是花生四烯酸的主要代謝產物之一,其在調控血管的舒張與抑制血小板凝集作用中扮演重要角色。雖然目前已知IP receptor可以活化Gs-PKA,進一步增加cAMP,但其相關訊息還有許多不清楚的地方,而為要進一步釐清,我們以human erythroleukemia cells (HEL) 為實驗模型,並選擇在臨床上用於治療肺性高血壓且具 IP receptor 專一性 prostacyclin 致效劑 beraprost 作為研究工具 。
IP receptor 為G-protein couple receptor (GPCR) 的一員,已知GPCR可以經由不同G-protein subunits和二級訊息來活化 MAPK pathway,因此我們便想瞭解IP receptor下游相關訊息傳遞為何?結果發現beraprost可以造成ERK1/2磷酸化,且是具有劑量及時間依賴性。在100nM時所造成ERK1/2 磷酸化則同時具 PKC 、PKA及PI3K依賴,因此便想繼續探討beraprost 所造成ERK1/2磷酸化其上游及下游可能的訊息傳遞。結果顯示beraprost 可以活化傳統MAPK pathway (RAF/MEK/ERK/ RSK) 。而由於PI3K 抑制劑(Wortmannin)可以降低beraprost所活化ERK1/2,因此我們便繼續探討beraprost與PI3K pathway 間的關係,結果發現beraprost 也可以活化PI3K/AKT/GSK3β pathway。由於我們也發現beraprost 所造成GSK3βphosphorylation 為MEK-dependent,因此認為在beraprost 所活化的MAPK 及PI3K pathways 中具有交互作用。
由上面結果顯示,IP agonist(beraprost)除了目前已知的訊息傳遞路徑外,還可以活化 MAPK 及 PI3K這兩條對細胞功能很重要的pathways。
英文摘要 Prostacyclin is one major metabolite of arachidonic acid and plays important roles in vascular dilation and platelet aggregation. Although prostacyclin receptor (IP) is known to activate Gs- protein kinase A (PKA) and then increases cAMP production, many questions about its signal transduction pathways remain to be answered. In order to study the signal transduction pathway involved in IP receptor activation, human erythroleukemia cells (HEL) and IP receptor specific agonist, beraprost, which is clinically for pulmonary hypertension, are used in this study.
In this study, 100 nM beraprost activates ERK1/2 phosphorylation in dose- and time-dependent manners which are Protein kinase C- (PKC), phosphatidylinositol 3-kinase- (PI3K), and PKA-dependent. When the upstream and downstream signals of ERK1/2 are further investigated, it is found that beraprost activates the MAPK (mitogen-activated protein kinase) pathway (RAF/MEK/ERK/ RSK). Since beraprost mediated ERK1/2 activation is also inhibited by PI3K inhibitor (Wortmannin), the relationship between beraprost and PI3K pathway is studied. It is found that beraprost also activates PI3K/AKT/GSK3pathway. For the reason that beraprost induced GSK3 phosphorylation is MEK-dependent, it is possible that beraprost induced MAPK and PI3K pathways might interact.
In conclusion, IP agonist (beraprost) activates both MAPK and PI3K pathways in addition to other known signal transduction in HEL cells.
論文目次 Contents

Contents Ⅲ
List of Figures Ⅳ
中文摘要 1
Abstract 3
Introduction 4
Materials and methods 13
Results 31
Discussion 41
Figures 46
References 68
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