在全世界中糖尿病腎病變為主要導致末期腎臟病的原因之一，且致死率於近年來不斷地攀升。過去的研究指出高血糖誘導的腎臟受損，活性氧物種(自由基)扮演一個重要的角色。嘉寶果，俗稱樹葡萄，目前多被研究是否能成為機能性食品，因富含花青素具有抗氧化的及抗發炎的特性。本研究旨在探討嘉寶果萃取物(MCE)是否能減緩第二型糖尿病小鼠的腎病變發展。首先，將 MCE 藉由分光光度計及高液相層析儀分析花青素及多酚的組成。在動物實驗方面，將 C57BL/6J 小黑鼠以 100 毫克/公斤之鏈佐黴素(streptozotocin, STZ)與 240 毫克/公斤之菸鹼胺(Nicotinamide, NA)注射腹腔並餵予高脂飼料誘導糖尿病及給予各種濃度的 MCE(0.1,0.5,1 %)。於每兩個禮拜監測一次血糖濃度、體重、葡萄糖耐受性測試，胰島素耐受性測試、血壓、腎臟超音波；犧牲後做腎臟切片分析 H&E 染色、PAS 染色、MASSON 式染色及免疫組織化學染色(IHC)。結果顯示，MCE 能穩定第二型糖尿病小鼠的血糖代謝並間接證實能改善胰島素敏感性，除此之外，糖尿病
腎病變導致的腎臟重量減輕、腎絲球萎縮、醣類堆積及膠原蛋白增生等現象，在 MCE 的給予之下也有所改善。在細胞實驗部分，利用高糖誘導 MES 13 腎絲球間質細胞增生，再觀察 MCE 對其抑制作用與相關機制。結果指出 MCE 可透過調節 Ras/PI3K/Akt 路徑而抑制腎絲球腎間質細胞增生，同時可減少ollagen IV 及 Fibronectin 等纖維化相關因子。本實驗證實 MCE 能延緩與改善糖尿病腎病變的病程，未來期待可進一步發展為一保健食品。

Diabetic nephropathy (DN) is a major cause of end-stage renal disease (ESRD) and the mortality is continuously progressing worldwide. Previous studies indicate that reactive oxygen species play an important role in high glucose-induced renal injury. Myrciaria cauliflora has been reported as a functional food rich in anthocyanins possessing anti-oxidative and anti-inflammative properties. This study examined whether Myrciaria cauliflora extracts (MCE) can attenuate diabetic nephropathy progression in type 2 DM mice. First, the composition of anthocyanins and polyphenols of MCE were determined by HLPC and spectrophotometer. One hundred mg/kg of streptozotocin and two hundred forty mg/kg nicotinamide were administered to C57BL/6J mice fed with high fat diet and variant concentrations of MCE. The plasma glucose concentration, body weight, oral glucose tolerance, insulin tolerance, blood pressure, renal ultrasoundultrasonic wavewere monitored every other two weeks. After sacrifice, the kidneysof mice were analyzed by using H&E, PAS, MASSON and IHC staining.The results showed that MCE stabilized the plasma glucose and indirect improved the insulin sensitivity in type 2 DM mice. In addition, diabetes-caused glomerular atrophy, accumulation of saccharide, andproliferation of collagen IVwere recovered or reduced under treatment with MCE in diabetic mice. Our results indicate that MCE has beneficial effects in diabetic nephropathy and the mechanism has been confirmed in vitro which inhibit proliferation via Ras/PI3K/Aktand kidney fibrosis related proteins.Comprehensive, this work will provide the potentialfor MCE asa natural food inhibiting the diabetic nephropathy.

壹、 縮寫檢索表 1
貳、 中文摘要 3
參、 英文摘要 5
肆、 文獻探討 7
4-1. 糖尿病簡介 7
4-1-1. 糖尿病之定義與診斷標準 7
4-1-2. 糖尿病之分類 7
4-1-3. 糖尿病之症狀 10
4-1-4. 糖尿病之併發症 12
4-2. 腎臟與糖尿病腎病變 16
4-2-1. 腎臟之簡介 16
4-2-2. 糖尿病腎病變之病程分為五個時期 17
4-2-3. 糖尿病造成腎病變之機制 19
4-3. 糖尿病動物模式 24
4-3-1. 自發型 24
4-3-2. 誘發型 25
4-4. 嘉寶果簡介 28
4-4-1. 背景介紹 28
4-4-2. 產地來源 28
4-4-3. 所含成分 28
4-4-4. 功效應用 29
伍、 研究動機 30
陸、 研究架構 31
柒、 實驗材料與方法 32
7-1. 實驗材料 32
7-2. 實驗方法 37
7-2-1. 嘉寶果萃取物的製備(MCE) 37
7-2-2. MCE定性分析 38
7-2-3. 細胞培養(cell culture) 38
7-2-4. 細胞毒性試驗(cytotoxicity assay) 39
7-2-5. MES13細胞增生試驗 (cell proliferation assay) 40
7-2-6. 西方點墨法 (western blotting) 40
7-2-7. 統計分析 42
7-2-8. 實驗動物 42
7-2-9. 第二型糖尿病(Type 2 DM)動物模式建立與實驗設計 43
7-2-10. 飼料組成與配置 43
7-2-11. 動物血糖之監測 45
7-2-12. 動物檢體之收集 45
7-2-13. 組織病理切片及染色 46
7-2-14. 血清中血脂分析 49
7-2-15. 血清中腎功能指標分析 49
7-2-16. 統計分析 50
捌、 實驗結果 51
8-1. MCE之定性與定量 51
8-2. 誘發第二型糖尿病(Type 2 DM)之實驗動物模式 52
8-3. 高脂質飼料餵養增加Type 2 DM C57BL/6J小黑鼠之體重 52
8-4. MCE具有穩定Type 2 DM C57BL/6J小黑鼠之血糖代謝的效果 52
8-5. MCE對於Type 2 DM C57BL/6J小黑鼠之血液生化值的影響 56
8-6. MCE具有降低Type 2 DM C57BL/6J小黑鼠之血壓的功效 57
8-7. MCE具有保護Type 2 DM C57BL/6J小黑鼠之腎臟的作用 57
8-8. MCE對於Type 2DM C57BL/6J小黑鼠之腎臟大小的影響 59
8-9. MCE對於MES13之毒性試驗 60
8-10. 在高糖狀態下MCE抑制MES13的增生 60
8-11. 在高糖狀態下MCE抑制MES13增生之相關的蛋白表現 61
玖、 討論 63
壹拾、 參考文獻 67
壹拾壹、 實驗結果圖表 74

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