2017-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656519摘要：代謝綜合徵（MetS）已經成為⼀個世界上重要的臨床問題，在台灣也是。MetS患者有較⾼的風險發展2 型糖尿病（T2DM）和⼼⾎管疾病（CVD）。胰島素抵抗（IR）是MetS 的核⼼問題，開發直接量測IR 的臨床診斷⼯具將可促進MetS 發病機制的研究並協助MetS 患者控制病情。脂肪組織（AT）是重要的儲存庫且可釋放許多調節代謝的激素，因此被認為是形成IR 的早期指標。脂肪細胞功能障礙的幾個重要病理節點是脂肪細胞肥⼤、缺氧、炎症和IR。螢光⽣命週期顯微術（FLIM）可測量活體內細胞的代謝狀態，應⽤於脂肪細胞的研究中將有潛⼒發展為體內檢測IR 的⼯具。這項研究的⽬的是在不同的代謝壓⼒環境下，包括脂肪細胞肥⼤、缺氧和炎症，驗證脂肪組織的FLIM 變化。我們的量測⽅法包含：使⽤FLIM 來評估脂肪細胞缺氧引起的糖酵解作⽤，並使⽤⾦奈⽶胰島素和2-NBDG 來分別確認胰島素和糖的攝取，並從圖像中分析脂肪細胞的肥⼤狀況。此外，使⽤巨噬細胞上標記EGFP 的基因轉殖⼩鼠中可以同時觀察脂肪細胞的炎症發⽣狀況。本計畫將進⾏細胞和組織研究，在不同的代謝壓⼒環境下的脂肪細胞驗證我們的量測⽅法。將進⾏動物研究，在螢光轉殖巨噬細胞的 C2J ⼩鼠上誘發糖尿病，驗證我們的量測⽅法。同時進⾏臨床研究，在CAD/DM 患者的脂肪組織切⽚上，驗證FLIM 的量測結果與CAD/DM 的關聯性。根據相關⽂獻調查以及我們先前的研究成果，我們預期FLIM 量測結果將可分辨出不同代謝壓⼒下的脂肪細胞，並在臨床研究上與CAD/DM 有顯著關聯性。故本計畫將成功建⽴⼀個診斷MetS 的新量測⽅法，有助於釐清脂肪細胞失調的上游機制並監控MetS 的發展過程。<br> Abstract: Metabolic syndrome (MetS) has been a major clinical issue worldwide, also inTaiwan. MetS patients have high risk to develop type 2 diabetes mellitus (T2DM) andcardiovascular disease (CVD). Insulin resistance (IR) is the core issue of MetS,developing a clinical diagnostic tool for direct measurement of IR will promote thestudy of MetS pathogenesis and help MetS patients control disease progression.Adipose tissue (AT) is the main repository which can release many hormonesregulating metabolism, thus regarded as an indicator of early development of IR. Themain pathological nodes of adipocyte dysfunction are adipocyte hypertrophy,hypoxia, inflammation, and IR. Fluorescence lifetime microscopy (FLIM) is apowerful tool to measure cell metabolism status in vivo. Applying FLIM in the studyof adipocytes will have the potential to develop a powerful tool for detecting IR invivo. The purpose of this study is to verify the FLIM measurement of AT in differentmetabolic stress status, including adipocyte hypertrophy, hypoxia, and inflammation.We will use FLIM to evaluate the glycolysis induced by hypoxia in adipocytes, anduse insulin-Au and 2-NBDG to confirm the uptake of insulin and sugar, and analyzethe hypertrophy of the adipocytes from the images. In addition, inflammation of theadipocytes can be observed simultaneously in mice that are transfected with EGFP onmacrophage. We will carry out cell and tissue studies to verify our measurements ofadipocyte in different metabolic stress status. Then, we will carry out animal studiesthat verify our measurements of AT of LyxM-EGFP C2J mice with DM. In parallel,we will carry out clinical studies that verify FLIM measurement of AT of patientswith CAD/DM. Based on the literature review and our previous work, we expect thatFLIM measurements will identify adipocytes in different metabolic stress status andwill be significantly associated with DM / CAD in clinical studies. Therefore, thisproject will successfully establish a novel measurement method for MetS diagnosis,benefit to clarify the upstream mechanism of adipocyte dysfunction and monitor thedevelopment of MetS.