臺灣博碩士論文加值系統

　糖尿病為全世界最嚴重的慢性疾病，也是造成末期腎臟疾病最主要的原因。在台灣，糖尿病的患者逐年攀升，而近年來末期腎臟疾病的盛行率以及發生率更攀升為世界第一，使得糖尿病與其併發症所造成的死亡人數直逼惡性腫瘤，耗費健保資源龐大。臨床上為了延緩尿毒症患者走向末期腎臟疾病的時程，會被建議以低蛋白飲食控制腎臟功能的衰竭病徵，吉多利錠(ketosteril)即為在低蛋白飲食下的一種必需胺基酸補充藥品，在動物以及人體試驗中皆能有效減緩尿毒症以及腎功能衰竭，但在糖尿病性腎病變中的研究報告較少。國內外有關腎臟病變的研究，包含糖尿病或非糖尿病所誘發之腎毒性動物模式，除了昂貴的基因缺陷型，如KK/HIJ 小鼠或是使用脂多醣的化學藥劑誘導型以外，都沒有便利且具實用性的糖尿病性腎病變動物模式，來提供研究者進一步探討糖尿病性腎病變後期的病理進程，如腎纖維化之形成的分子機轉等。
　　牛樟芝，或稱樟芝或牛樟菇，為台灣特有的一種真菌，在現代醫學的應用廣泛已被大量研究發表，其所含有的活性成分在動物以及細胞模式中證實具有抗纖維化、抗氧化、抗發炎以及降血糖的功效，正是近幾年糖尿病性腎病變的研究報告中，被討論的主要治療途徑。近幾年的研究報告已證實牛樟芝具有治療慢性腎臟疾病的潛力，可抑制血中尿毒素的上升並降低蛋白尿的排泄量，並觀察到可減少腎絲球基底膜的增厚情形。目前國內外關於牛樟芝的疾病研究，仍以癌症以及肝纖維化等主軸占多數，對於腎臟病變方面的研究仍寥寥可數，更無人探討牛樟芝對於糖尿病性腎病變的生理活性，因此，探討牛樟芝對於糖尿病性腎病變的治療作用以及其機轉，是一個具前瞻性的研究議題。
　　本論文利用低劑量的STZ誘導方式，合併高脂質飼料的選擇性給予，建立一個便利且具實用性之第二型糖尿病性腎病變動物模式，並將其分別應用在探討低蛋白飲食合併吉多利錠以及牛樟芝子實體萃取物改善第二型糖尿病性腎病變之藥理活性。在低蛋白飲食合併吉多利錠研究中，我們發現吉多利錠可經由抑制乙型轉型生長因子(TGF-β) 以及腫瘤壞死因子(TNF-α)之機轉，有效改善第二型糖尿病性腎臟病變情形。此外，在牛樟芝合併低蛋白飲食研究中，第二型糖尿病性腎病變小鼠經由牛樟芝子實體萃取物投予後，發現單獨餵食牛樟芝即可藉由降低血清中葡萄糖濃度、改善血中脂質代謝調控以及減少尿蛋白排出量的方式，有效改善腎間質堆積情形。利用免疫組織染色法，我們證實牛樟芝可藉由降低腎纖維化因子TGF-β以及α-SMA的表現及分佈，來達到抑制腎臟基質增生及後期纖維化之功效，顯示牛樟芝不需要配合低蛋白飲食，即有顯著提升糖尿病小鼠腎臟功能及抑制進行性腎病變之藥理活性，值得進一步去評估開發治療糖尿病性腎病變之新用途。

Diabetic nephropathy (DN) is one of the major diseases that results in chronic kidney disease (CKD). It is also a common and serious complication of diabetes mellitus, which leads to renal failure in up to 30% of individuals with diabetes and thus becomes the most important end-stage renal diseases (ESRD). In the study, we look for constructing several kinds of non-genetic deficient diabetic nephropathy (DN) mouse models. It is suggested that disease models prior to serving with renal fibrosis, which may be utilized for advanced studies on DN animal models. A nongenetic model of type 2 diabetes has been described that involves feeding rats a high fat diet and administering a low dose of STZ. Therefore, it is important to establish a strain-appropriate STZ dosage protocol for achieving hyperglycaemia before embarking on a major study of diabetic nephropathy using this model. Restricting dietary protein is the one of major components of therapy in CKD patients and aims to slow the progression of renal failure and to provide optimum nutritional status. Many authors have demonstrated that low-protein diet (LPD) and very low-protein diet (VLPD) mixture of EAA ketoanalogs, which are used to prevent nutritional deficiencies caused by protein-restricted diets. It contains insulin resistance, hyperlipidemia and delay the start of renal replacement.
Antrodia camphorata (AC) is a unique fungal species that used as a folk medicine, has attracted extreme attention for inflammation syndromes and liver-related diseases research in Taiwan. It has been known that AC can play the role on anti-oxidation, has recently been marketed in the forms of nutraceuticals. It has been suggested that AC protects the kidney from auto-immune disease. However, the ability of AC to DN has not been subjected to scientific scrutiny.
This study investigated the non-genetic deficient DN animal models, which closely simulates the metabolic abnormalities of the human disease, and it is also been available by cost-effective. The hyperglycemia and hyperinsulinaemia were induced on these HFD-controlled C57BL/6 mice (fasting blood glucose over 180 mg/dl, and ACR over 60 g/mg, alternatively). We also evaluated the metabolic profiles, renal histology and the mechanism of AC treated mice in DN models. We found that the AC has improve the glucose tolerance and increasing blood high density lipoprotein-low density lipoprotein ratio. Immunohistochemical evaluation also showed weak TGF-β and α-SMA staining in the renal glomeruli of native and AC group, while the signal revealed markedly stronger in the control group. In conclusion, our data shows a new non-genetic deficient DN animal models, and prove that the alcoholic extract from AC treatment has shown a potential anti-fibrosis effect in HFD-induced diabetic nephropathy mice.

總目錄
中文摘要……………………………………………………………………………….I
英文摘要…………………………………………………………………………..…...II
圖及表目錄…………………………………………..………………………………...V
縮寫對照表…………………………………………………………………………….VII
一、 緒論……………………………………………………………………………..1
二、 文獻回顧…………………………………………….………………………….4
1. 糖尿病性腎病變之研究
1.1糖尿病性末期腎臟病變......................................................................................4
1.2高血糖與糖尿病性腎臟病變……………………………………….………….5
1.3高度糖化終產物與糖尿病性腎臟病變………………………………..............6
1.4氧化壓力與糖尿病性腎臟病變………………………………………..……....8
1.5蛋白尿與糖尿病性腎臟病變……………………………………….…..…..….9
1.6糖尿病性腎病變之分子標靶及機制………………………….….………..….10
1.7糖尿病性腎病變小鼠模式建立……………………………………..….……..13
2. 糖尿病腎病變治療藥物及其機轉
2.1胰島素增敏劑………………………………………………………..………...16
2.2吉多利錠( ketosteril )………………………………………….…...………..…...18
2.3牛樟芝在現代醫學的應用潛力……………………………….………..……..19
三、 研究動機與目的…………………………………………….…..……. ……....23
四、 實驗材料與方法…………………………………………….……....................24
4.1實驗材料……………………………………………………….…….....….…..24
4.2儀器設備…………………………………………..…………………………...25
4.3不同STZ濃度與HFD給予誘導糖尿病性腎病變動物模式研究………......26
4.4吉多利錠合併低蛋白飲食改善糖尿病性腎病變機制研究…………….…....29
4.5牛樟芝合併低蛋白飲食改善糖尿病性腎病變機制研究………………….....30
五、 結果與討論………………………………………………………………….....35
5.1不同STZ濃度與HFD給予誘導糖尿病性腎病變動物模式研究……….....35
5.2吉多利錠合併低蛋白飲食改善糖尿病性腎病變機制研究…………...….….37
5.3牛樟芝合併低蛋白飲食改善糖尿病性腎病變機制研究.................................39
六、 結論…………………………………..………………………………….……...43
七、 結果圖表………………………………………………………………….…….44
八、 參考文獻…………………………………………………………………….….73

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