2010-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/660545摘要：研究目的：目前對於代謝疾病導致動脈硬化的成因、發生與否、以及預後，了解仍有限。因此，利用生物標記找出高危險群並預測預後，對於臨床照顧非常重要。初步研究顯示，血中vascular adhesion protein-1 (簡稱VAP-1，不是vascular cellular adhesion molecule-1，也不是VCAM-1)是血中氧化壓力與advanced glycation end products 的來源之一，會參與發炎與動脈硬化過程，我們更首先發現文獻中不曾報告過的細胞－單核球與巨嗜細胞也會表現VAP-1。本計畫將探討發炎因子，特別是VAP-1，在代謝與動脈硬化疾病扮演的角色及可能的應用。本計畫包括兩方面。第一方面是生物標記的研究：我們將探討血清中VAP-1 在臨床上是否可以做為代謝與動脈硬化疾病的生物標記，包括頸動脈狹窄，週邊血管阻塞疾病、以及糖尿病神經病變(specific aim a)，也將研究其他發炎因子在口服葡萄糖耐受試驗中的變化，是否是動脈硬化更好的生物標記(specific aim b)。另一方面是致病機轉的研究：我們將於細胞模式探討巨嗜細胞表現的VAP-1 在發炎與動脈硬化的過程有何功能，以及可能的調節因子(specific aim c)，也將研究單核球與巨嗜細胞是否是血清中VAP-1 的來源之一(specific aim d)。研究設計與方法：將以前瞻性病例對照研究法收錄頸動脈狹窄的病人，記錄臨床資料並測量血中VAP-1 濃度，以探討血中VAP-1 是否與頸動脈狹窄有關，並研究氣球擴張術或是置放支架前後，血中VAP-1 濃度的變化是否可以預測預後。其次，針對在台大醫院長期追蹤的糖尿病患，利用縱斷式世代研究法，研究血中VAP-1 濃度是否可以預測這些病患未來發生週邊血管阻塞疾病的機會。接著，採用病歷對照研究法，探討血中VAP-1 濃度與糖尿病神經病變的關係，並分析血中VAP-1 濃度是否可以做為施行神經切片與否的依據。此外，也將研究血清中發炎因子的濃度，包括intercellular adhesion molecule-1、monocyte-chemotactic protein-1、E-selectin、CD-40 ligand、以及C-reactive protein，是否會在口服葡萄糖水後產生變化？發炎因子的變化與冠狀動脈心臟病及嚴重度是否有關？致病機轉方面，將利用THP-1 人類單核球細胞株，加入調節因子TNF-α，葡萄糖與胰島素，比較巨嗜細胞表現VAP-1 的變化。此外，也將運用RNA interference 技術，比較VAP-1 knock-down 後，巨嗜細胞表現附著因子與受器的量、分泌酵素與cytokines 的濃度、以及氧化低密度脂蛋白的能力是否有所不同。此外，將測定培養液中VAP-1 的濃度是否受前述調節因子所調控；並運用正子攝影，探討動脈硬化斑中的巨嗜細胞數與血清中的VAP-1 的關係；再收集人類週邊血液單核球，分析其VAP-1 mRNA 表現量與血清中VAP-1 濃度是否有關；以確定單核球與巨嗜細胞是否是血清中VAP-1 的來源之一。<br> Abstract: Objective: The understanding of how metabolic abnormality leads to atherosclerosisremains limited. Therefore, to develop biomarkers to identify high-risk groups and to predictprognosis is important clinically. Our preliminary results showed that vascular adhesionprotein-1 (VAP-1, not vascular cellular adhesion molecule-1, not VCAM-1) is a sourceof systemic oxidative stress and advanced glycation end products.We also found thatmonocyte/macrophage expresses VAP-1, which can participate in inflammation andatherosclerosis.We will explore the role and the applications of inflammatory mediators,especially VAP-1, in metabolic and atherosclerotic diseases in this project.Firstly, we will explore if serum VAP-1 could be a useful biomarker for metabolic andatherosclerotic diseases, including carotid arterial stenosis, peripheral arterial occlusivedisease, and diabetic neuropathy (specific aim a). We will also study the dynamic change ofinflammatory markers during oral glucose tolerance test, and whether this change is betterthan fasting levels in predicting atherosclerosis (specific aim b). Secondly, we will study thepathogenesis of atherosclerosis.We will explore the potential function and regulators ofVAP-1 on macrophage. (specific aim c).Whether monocyte and macrophage are sources ofsystemic VAP-1 will also be studied (specific aim d).Research design and methods: Firstly, we will include patients with carotid arterialstenosis and analyze if serum VAP-1 is associated with carotid arterial stenosis usingprospective case-control design.We will study if change of serum VAP-1 duringpercutaneous transluminal angioplasty can predict outcome after the procedure. Secondly,subjects with diabetes followed at our hospital for a long time will be enrolled in alongitudinal cohort study.We will explore if serum VAP-1 can predict the occurrence ofperipheral arterial occlusive disease in the future. Thirdly, using case-control design, we willsee if serum VAP-1 is associated with diabetic neuropathy, and whether serum VAP-1 can beused to decide if nerve biopsy is indicated in these subjects.We will also test if inflammatorymarkers, including intercellular adhesion molecule-1, monocyte-chemotactic protein-1,E-selectin, CD-40 ligand, and C-reactive protein change during oral glucose tolerance test.Analyses of whether these changes are associated with coronary arterial disease and itsseverity will be done.We will add TNF-α, glucose, and insulin to see the change of VAP-1 expression inhuman monocytic cell line THP-1.We will also knock-down VAP-1 on macrophage usingRNA inference technology, to compare the expression of adhesion molecules and receptors,表C011 共2 頁第2 頁the secretion of enzymes and cytokines, and the ability to oxidize low-density lipoprotein.Meanwhile, we will check if the regulators can change the secretion of VAP-1 to culturemedia.We will correlate serum VAP-1 to the aggregation of macrophage in atheroscleroticplaque using PET.We will also analyze the association between VAP-1 mRNA expression ofhuman peripheral monocyte and serum VAP-1. Through these, we want to prove thatmonocyte and macrophage are sources of systemic VAP-1.