2018-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/699537摘要：本計劃曾連續兩年提出但都未獲通過，審查意見認為應加強臨床試驗部份及preliminary data，也建議動物實驗。今年我們持續補強計劃内容，重新提出申請，希望能得到審查委員的肯定。(1) Preliminary data：我們增加了許多創新的研宄，絕對不是me-too study !這是國際間第一個探討負電性L5-LDL在急性心肌梗塞發炎反應之研宄，更是第一個探討L5-LDL與Interleukin 1(3、G-CSF及GM-CSF臨床相關性之研宄。初步data已寫成論文發表，我們有信心一定可繼續得到好結果。(2)臨床試驗：負電性L5-LDL與急性心肌梗寒STEMI發炎基因相關性之臨床試驗更是全球首仞，有很高的臨床應用價值。(3)動物實驗：可以提供更多in vivo資料來支持L5-LDL與血管發炎反應的關連。急性心肌梗塞是全球死亡率最高的急症之一，其機轉根源於動脈硬化損傷的形成，而低密度脂蛋白 (low-density lipoprotein, LDL)的氧化據信是一重要致病因素。近年來我們發現，急性心肌梗塞病人 血液中一個負電性的氧化L5-LDL大量增加。同時，病人的發炎細胞激素Interleukin (IL)-1(3也顯著上升。然而，L5-LDL與發炎反應的相關性，以及發炎激素宄竟是否經由L5-LDL所誘導仍不清楚。我們提出這個研宄計劃目的為釐清L5在急性心肌梗塞發炎所扮演的角色。因此，我們要分析急性心 肌梗塞病人血液中L5與發炎細胞激素IL-1(3之濃度，並釐清L5是否可促使IL-1(3的分泌。由於在動脈 損傷區域會有嗜中性白血球和它所產生的介質存在，例如：顆粒性白血球群落刺激因子（granulocyte colony-stimulating factor, G-CSF),及顆粒球巨■細胞群落刺激因子（granulocyte macrophage colony-stimulating factor, GM-CSF),所以 G-GCF 及 GM-CSF 也是研宄重點。我們預定以兩年的時間來進行這項計劃，首先在單一醫學中心執行至少兩年臨床試驗確立L5-LDL與 IL-1P, G-GCF及GM-CSF的關連。另外，本計劃的第一年將探討急性心肌梗塞患者的L5-LDL是否能夠 造成人類巨噬細胞中IL-1(3、G-CSF和GM-CSF的表現增加，初步實驗結果已經支持這項假設。本計 劃的第二年，將研宄此帶負電的L5-LDL如何引起這些發炎反應。我們會分離出人類巨噬細胞做實驗， 利用抑制劑和shRNA knockdown的方法，檢視L5-LDL誘導G-CSF和GM-CSF的訊息傳遞機制，特 別是NF- k R ERK2及JNK的角色。另外，我們也將採用心肌梗塞的動物實驗模式，進一步了解L5-LDL 與這些inflammatory cytokines的相關性。創新性：這個計劃包括許多創新的研究，是全球第一個探討負電性L5-LDL在急性心肌梗塞發炎反應 基因調控之研宄，更是國際間第一個探討L5-LDL調控Interleukin 1(3、G-CSF及GM-CSF引起血管發炎反應之研宄。其結果將有助於我們瞭解負電性LDL如何造成心血管疾病的發炎反應，藉由這些認 識我們也希望可以發展出早期診斷急性心肌梗塞的生物試劑。<br> Abstract: Acute myocardial infarction originates from atherosclerotic injury of the vessel walls where oxidized LDL and monocyte-derived macrophages are accumulated. Electronegative low-density lipoprotein (LDL) is a key risk factor for atherosclerotic disease, but the precise mechanism of electronegative LDL，s contribution to atherosclerosis remains elusive. We have identified an increase of a negatively charged LDL species named L5-LDL, along with an increase of Interleukin (IL)-ip, in patients with acute myocardial infarction. However, the role of L5-LDL in vascular inflammation and IL-ip cytokine regulation is not known.The purpose of the proposed studies is to investigate the role of electronegative L5-LDL in the vascular inflammation in acute myocardial infarction, with emphasis on the inflammatory response in macrophages. We will measure plasma levels of L5 and an inflammatory cytokine, interleukin 1(3，in patients with acute ST-elevation myocardial infarction (STEMI) during acute phase and follow-up. Neutrophil-related granulocyte colony-stimulating factor (G-CSF) and granulocyte macrophage colony-stimulating factor (GM-CSF) will also be investigated. Mechanisms underlying the regulation of IL-1(3，G-CSF and GM-CSF gene expression by electronegative L5-LDL are a primary goal of this project.This project will be performed in a 2-year time frame, during which a STEMI clinical trial will be continued without interruption to establish the clinical correlation of L5-LDL and IL-ip, G-CSF and GM-CSF in STEMI. In the first project year, we will also investigate whether L5-LDL can regulate the expression of G-CSF, GM-CSF and IL-ip. In the second year, we will elucidate the molecular mechanism of L5-induced inflammatory response in human macrophages. Signal pathway modifiers and shRNA knockdown will be used to evaluate the role of transcription factors such as NF-kB, ERK2 and JNK. Animal studies will also provide in vivo data on the relationship of L5-LDL and inflammatory cytokines.Results of this study will be the first to demonstrate inflammatory cytokines regulation by L5-LDL, and also the first to unravel the molecular mechanism underlying L5-LDL-induced vascular inflammation in acute myocardial infarction. These data can further our understanding of electronegative LDL-induced vascular inflammation，enabling the development of early diagnostic agents for acute myocardial infarction.急性心肌梗塞負電性LDLIL-1(3顆粒性白血球群落刺激因子(G-CSF)顆粒球巨噬細 胞群落刺激因子(GM-CSF)巨噬細胞發炎反應Acute myocardial infarction • Electronegative LDL • Interleukin-1(3 • G-CSF • GM-CSF • Macrophages • Inflammation