2012-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/658660摘要：研究目的：根據我們先前的研究發現，以傳統的危險因子來預測糖尿病的發生，仍會錯失50%的高危險群，因此我們希望利用血清中生物標記的測定，來改善高危險群的定義。因此，本研究的第一部份將尋找可預測糖尿病與代謝症候群發生的生物標記，包括vascular adhesion protein-1(簡稱VAP-1，不是vascular cellular adhesionmolecule，也不是VCAM-1)，以及運用代謝體方法(metabolomics)尋找的小分子生物標記(specific aim 1A & B)。此外，也將建立一5 年追蹤世代，以探討糖化血色素(hemoglobinA1c)與新發生的糖尿病、腎臟病及頸動脈狹窄的關係，試圖提供國人糖化血色素定義糖尿病與糖尿病高危險群的數據基礎(specific aim 1C)。本計畫第二部分是研究VAP-1 在肥胖與糖尿病致病機轉的角色。首先是觀察性研究：我們將分析人類大網膜(omental)與皮下(subcutaneous)脂肪組織VAP-1 表現量與肥胖及胰島素阻抗的關係，並且探討人類血中VAP-1 濃度、內臟脂肪量與皮下脂肪量的關係(specific aim 2A & 2C)。由於肥胖導致糖尿病的部分原因，是透過降低局部adiponectin 與升高monocyte chemotatic protein-1 (MCP-1)的機轉，所以我們也將分析人類脂肪組織VAP-1、MCP-1 與adiponectin 表現量的關係(specific aim 2A)。接著是半介入性研究：隨著時間經過，有些人會變胖，有些人會變瘦，有些人的體型不變，胰島功能與胰島素阻抗也一樣。我們將研究人類肥胖程度、胰島功能與胰島素阻抗隨著時間的改變，是否伴隨著血中VAP-1 濃度的變化，以得知兩者的關係(specific aim 2D)。最後是介入性研究，我們將比較糖尿病人接受rosiglitazone 改善胰島素阻抗與血糖後，血中VAP-1 濃度是否有變化(specific aim 2E)，並且在細胞模式中，研究VAP-1 是否可以調控脂肪細胞與巨噬細胞adiponectin 與MCP-1 的表現(specific aim 2B)。研究設計與方法：Specific aim 1A 將以世代研究法(cohort study)，以time-resolvedimmunofluometric assay 測量血中VAP-1 濃度，並分析血中VAP-1 濃度是否可以預測2年後發生糖尿病與代謝症候群。Specific aim 1B 採用巢式病例對照研究法(nestedcase-control study)，以LC-QTOF MS 及GC-TOF MS 測定代謝體，尋找可預測糖尿病與代謝症候群發生的代謝產物。Specific aim 1C 將針對先前追蹤的世代，透過問卷、身體檢查、抽血與尿液檢查，了解經過5 年之後這些民眾的健康狀態。Specific aim 2A 將針對接受一般外科常規手術的成人，以橫斷式研究法進行分析，收集受試者大網膜與皮下脂肪組織，萃取出蛋白質與RNA，以分析VAP-1、MCP-1 及adiponectin 的表現，並與臨床表徵做統計分析。Specific aim 2B 將針對3T3-L1 脂肪細胞與RAW 巨噬細胞，加入VAP-1 酵素活性的抑制劑或是recombinant mouse VAP-1 後，測定細胞MCP-1 與adiponectin 的mRNA、cellular protein 與protein secretion 是否改變。Specific aim 2C 將利用橫斷面研究法，測定人類血中VAP-1 濃度，並以電腦斷層檢查分析內臟脂肪與皮下脂肪的量，再分析三者的關係。Specific aim 2D 將分析隨著時間經過，人類血中VAP-1 濃度、肥胖程度、胰島功能與胰島素阻抗的關係。Specific aim 2E將以雙盲及placebo-control 研究，比較糖尿病人接受rosiglitazone 或是placebo 6 個月後，血中VAP-1 濃度的變化。<br> Abstract: Objective: To predict diabetes in the future by traditional risk factors, 50% of high-risksubjects will not be identified, based on our preliminary results. Therefore, this project aimsto improve the definition of high-risk group by serum biomarkers. The first part of thisproject is to search for potential serum markers which can predict the development ofdiabetes and metabolic syndrome in the future, including serum vascular adhesionprotein-1 (VAP-1, not vascular cellular adhesion molecule, not VCAM-1) and serummetabolites found by metabolomics (specific aim 1A& 1B). Besides, we will also establisha cohort which has been followed for 5 years, to analyze the relationship of hemoglobin A1cto diabetes, chronic kidney disease, and atherosclerosis in carotid arteries in the future(specific aim 1C). This can provide evidence to support the definition of diabetes andpre-diabetes in Taiwan.The second part of the project is to investigate the role of VAP-1 in the pathogenesis ofobesity and diabetes.We will study the relationship between the expression of VAP-1 inomental and subcutaneous adipose tissue, obesity, and insulin resistance, and analyze therelationship between serum VAP-1 and area of visceral and subcutaneous fat in humansubjects (specific aim 2A & 2C). Since monocyte chemotactic protein-1 (MCP-1) andadiponectin is involved in the pathogenesis of diabetes, we will also explore the relationshipbetween the expression of VAP-1, MCP-1 and adiponectin in human adipose tissues(specific aim 2A). Besides, we will also analyze the change of serum VAP-1 duringfollow-up, and correlate with the change of obesity, insulin secretion function, and insulinresistance in human (specific aim 2D).We will study if serum VAP-1 changes in response torosiglitazone treatment in subjects with diabetes (specific aim 2E). In cell models, we willinvestigate if VAP-1 can regulate the expression and secretion of MCP-1 and adiponectin inadipocytes and macrophages (specific aim 2B).Research design and methods: In specific aim 1A, we will measure serum VAP-1 bytime-resolved immunofluometric assay to analyze if serum VAP-1 can predict thedevelopment of diabetes and metabolic syndrome in 2 years in a cohort study. In specificaim 1B, we will measure metabolites by LC-QTOF MS and GC-TOF MS, and search forpotential metabolites to predict diabetes and metabolic syndrome by nested case-controldesign. In specific aim 1C, we will follow a cohort at 5 years, and to know their health statusby questionnaires, physical examination, blood tests, and urinary tests.In specific aim 2A, subjects receiving scheduled surgery will by recruited in across-sectional study. Their omental and subcutaneous adipose tissue will be collected.Protein and RNA will be extracted. Expression of VAP-1, MCP-1, and adiponectin will beanalyzed and correlate with clinical characteristics. In specific aim 2B, inhibitors of VAP-1or recombinant mouse VAP-1 will be added to evaluate if VAP-1 can regulate the expressionand secretion of MCP-1 and adiponectin in 3T3-L1 adipocytes and RAW macrophages byautocrine or paracrine effect. In specific aim 2C, we will measure serum VAP-1 and computethe area of visceral and subcutaneous fat, and analyze their correlations in a cross-sectionalstudy.We will also analyze the relationship among change of serum VAP-1, obesity, insulinsecretion function, and insulin resistance by time in a cohort study (specific aim 2D). Inspecific aim 2E, we will investigate if serum VAP-1 changes by rosiglitazone in arandomized, placebo-controlled clinical trial in subjects with type 2 diabetes.研究目的：根據我們先前的研究發現，以傳統的危險因子來預測糖尿病的發生，仍會 錯失50%的高危險群，因此我們希望利用血清中生物標記的測定，來改善高危險群的 定義。因此，本研究的第一部份將尋找可預測糖尿病與代謝症候群發生的生物標記， 包括vascular adhesion protein-1(簡稱VAP-1，不是vascular cellular adhesion molecule，也不是VCAM-1)，以及運用代謝體方法(metabolomics)尋找的小分子生物標 記(specific aim 1A & B)。此外，也將建立一5 年追蹤世代，以探討糖化血色素(hemoglobin A1c)與新發生的糖尿病、腎臟病及頸動脈狹窄的關係，試圖提供國人糖化血色素定義 糖尿病與糖尿病高危險群的數據基礎(specific aim 1C)。 本計畫第二部分是研究VAP-1 在肥胖與糖尿病致病機轉的角色。首先是觀察性研 究：我們將分析人類大網膜(omental)與皮下(subcutaneous)脂肪組織VAP-1 表現量與肥 胖及胰島素阻抗的關係，並且探討人類血中VAP-1 濃度、內臟脂肪量與皮下脂肪量的 關係(specific aim 2A & 2C)。由於肥胖導致糖尿病的部分原因，是透過降低局部 adiponectin 與升高monocyte chemotatic protein-1 (MCP-1)的機轉，所以我們也將分析人 類脂肪組織VAP-1、MCP-1 與adiponectin 表現量的關係(specific aim 2A)。接著是半介 入性研究：隨著時間經過，有些人會變胖，有些人會變瘦，有些人的體型不變，胰島 功能與胰島素阻抗也一樣。我們將研究人類肥胖程度、胰島功能與胰島素阻抗隨著時 間的改變，是否伴隨著血中VAP-1 濃度的變化，以得知兩者的關係(specific aim 2D)。 最後是介入性研究，我們將比較糖尿病人接受rosiglitazone 改善胰島素阻抗與血糖後， 血中VAP-1 濃度是否有變化(specific aim 2E)，並且在細胞模式中，研究VAP-1 是否可 以調控脂肪細胞與巨噬細胞adiponectin 與MCP-1 的表現(specific aim 2B)。 研究設計與方法：Specific aim 1A 將以世代研究法(cohort study)，以time-resolved immunofluometric assay 測量血中VAP-1 濃度，並分析血中VAP-1 濃度是否可以預測2 年後發生糖尿病與代謝症候群。Specific aim 1B 採用巢式病例對照研究法(nested case-control study)，以LC-QTOF MS 及GC-TOF MS 測定代謝體，尋找可預測糖尿病 與代謝症候群發生的代謝產物。Specific aim 1C 將針對先前追蹤的世代，透過問卷、身 體檢查、抽血與尿液檢查，了解經過5 年之後這些民眾的健康狀態。 Specific aim 2A 將針對接受一般外科常規手術的成人，以橫斷式研究法進行分析， 收集受試者大網膜與皮下脂肪組織，萃取出蛋白質與RNA，以分析VAP-1、MCP-1 及 adiponectin 的表現，並與臨床表徵做統計分析。Specific aim 2B 將針對3T3-L1 脂肪細 胞與RAW 巨噬細胞，加入VAP-1 酵素活性的抑制劑或是recombinant mouse VAP-1 後， 測定細胞MCP-1 與adiponectin 的mRNA、cellular protein 與protein secretion 是否改變。 Specific aim 2C 將利用橫斷面研究法，測定人類血中VAP-1 濃度，並以電腦斷層檢查 分析內臟脂肪與皮下脂肪的量，再分析三者的關係。Specific aim 2D 將分析隨著時間經 過，人類血中VAP-1 濃度、肥胖程度、胰島功能與胰島素阻抗的關係。Specific aim 2E 將以雙盲及placebo-control 研究，比較糖尿病人接受rosiglitazone 或是placebo 6 個月 後，血中VAP-1 濃度的變化。Objective: To predict diabetes in the future by traditional risk factors, 50% of high-risk subjects will not be identified, based on our preliminary results. Therefore, this project aims to improve the definition of high-risk group by serum biomarkers. The first part of this project is to search for potential serum markers which can predict the development of diabetes and metabolic syndrome in the future, including serum vascular adhesion protein-1 (VAP-1, not vascular cellular adhesion molecule, not VCAM-1) and serum metabolites found by metabolomics (specific aim 1A& 1B). Besides, we will also establish a cohort which has been followed for 5 years, to analyze the relationship of hemoglobin A1c to diabetes, chronic kidney disease, and atherosclerosis in carotid arteries in the future (specific aim 1C). This can provide evidence to support the definition of diabetes and pre-diabetes in Taiwan. The second part of the project is to investigate the role of VAP-1 in the pathogenesis of obesity and diabetes.We will study the relationship between the expression of VAP-1 in omental and subcutaneous adipose tissue, obesity, and insulin resistance, and analyze the relationship between serum VAP-1 and area of visceral and subcutaneous fat in human subjects (specific aim 2A & 2C). Since monocyte chemotactic protein-1 (MCP-1) and adiponectin is involved in the pathogenesis of diabetes, we will also explore the relationship between the expression of VAP-1, MCP-1 and adiponectin in human adipose tissues (specific aim 2A). Besides, we will also analyze the change of serum VAP-1 during follow-up, and correlate with the change of obesity, insulin secretion function, and insulin resistance in human (specific aim 2D).We will study if serum VAP-1 changes in response to rosiglitazone treatment in subjects with diabetes (specific aim 2E). In cell models, we will investigate if VAP-1 can regulate the expression and secretion of MCP-1 and adiponectin in adipocytes and macrophages (specific aim 2B). Research design and methods: In specific aim 1A, we will measure serum VAP-1 by time-resolved immunofluometric assay to analyze if serum VAP-1 can predict the development of diabetes and metabolic syndrome in 2 years in a cohort study. In specific aim 1B, we will measure metabolites by LC-QTOF MS and GC-TOF MS, and search for potential metabolites to predict diabetes and metabolic syndrome by nested case-control design. In specific aim 1C, we will follow a cohort at 5 years, and to know their health status by questionnaires, physical examination, blood tests, and urinary tests. In specific aim 2A, subjects receiving scheduled surgery will by recruited in a cross-sectional study. Their omental and subcutaneous adipose tissue will be collected. Protein and RNA will be extracted. Expression of VAP-1, MCP-1, and adiponectin will be analyzed and correlate with clinical characteristics. In specific aim 2B, inhibitors of VAP-1 or recombinant mouse VAP-1 will be added to evaluate if VAP-1 can regulate the expression and secretion of MCP-1 and adiponectin in 3T3-L1 adipocytes and RAW macrophages by autocrine or paracrine effect. In specific aim 2C, we will measure serum VAP-1 and compute the area of visceral and subcutaneous fat, and analyze their correlations in a cross-sectional study.We will also analyze the relationship among change of serum VAP-1, obesity, insulin secretion function, and insulin resistance by time in a cohort study (specific aim 2D). In specific aim 2E, we will investigate if serum VAP-1 changes by rosiglitazone in a randomized, placebo-controlled clinical trial in subjects with type 2 diabetes.