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研究生:黃騰億
研究生(外文):Huang , Teng I
論文名稱:冬蟲夏草於糖尿病大鼠模式中之降血糖機制探討
論文名稱(外文):Studies on the Mechanism of Anti-hyperglycemic Activity of Cordyceps sinensis in Diabetic rats
指導教授:盧錫祺
指導教授(外文):Lu , Hsi Chi
學位類別:碩士
校院名稱:東海大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:108
中文關鍵詞:冬蟲夏草糖尿病高血糖葡萄糖轉運蛋白葡萄糖激酶胰島素受器
外文關鍵詞:Cordyceps sinensisdiabeteshyperglycemiaglucose transporterglucokinaseinsulin receptor
相關次數:
  • 被引用被引用:20
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  • 評分評分:
  • 下載下載:355
  • 收藏至我的研究室書目清單書目收藏:8
隨著生活水準不斷提高及醫療衛生的進步,國人平均壽命逐漸延長,老年人口持續增加,慢性疾病遂成威脅國人健康的重要疾病。近年來,糖尿病便開始進入國人十大死因排行榜上。針對糖尿病之保健食品不斷推陳出新,其中有許多天然物所含的多醣類物質,經動物實驗證實是具有降血糖的效用。在天然生成的冬蟲夏草中,由於含有高含量之多醣物質,可能有助於減緩高血糖之症狀。之前研究證實,採用 streptozotocin (STZ) 能成功運用在 Wistar 大鼠誘發糖尿病,餵食冬蟲夏草子實體之實驗組,發現有降低高血糖之顯著效果。本研究嘗試探究大鼠模式中,冬蟲夏草如何在不影響胰島素分泌量之下,達成降血糖之作用,分別針對葡萄糖的恆定機制與代謝路徑進行追蹤。 Wistar 大鼠經由 STZ 誘發糖尿病後,分別口服冬蟲夏草之子實體、蟲體,以及兩者之混合物;經分析肝臟中葡萄糖轉運蛋白 GLUT2 、葡萄糖激酶 GK 與胰島素受器 InR 表現量、GK 酵素活化性等參數。初步結果顯示,除 GK 外,口服冬蟲夏草的各組,其肝臟細胞 GLUT2 與 InR 表現均較未食用 STZ 控制組為高。GK 基因表現在蟲體或子實體分食時略降,而於合併給於時則增加,其意義有待探討。推測冬蟲夏草經由口服,可能藉更多的血糖轉運蛋白與胰島素接受器,以及更有效率的葡萄糖磷酸化以達成降低血糖之效應。
Diabetes mellitus is one of the life-threatening chronic diseases and has become a top-four killer in Taiwan(Department of health, 2004). Cordyceps sinensis is one of the most precious Chinese herbs and may play various physiological roles in restoring the body’s homeostasis. In addition to those well-known functions, recent evidences suggested that extracts of Cordyceps, most probably polysaccharide fractions, may have help to alleviate hyperglycemia in diabetic animals. However the underlying molecular mechanisms have not been addressed. In this study, we attempted to clarify how Cordyceps affect the hepatic glucose homeostasis in diabetic rats. Male Wistar rats were injected with streptozotocin for diabetes induction of diabetes and then were fed with carcass of Cordyceps, fruiting body, mixture of the fruiting body and carcass, and starch; that is, the CC, FB, CF and STZ groups, respectively. Hepatic glucokinase activities were determined from enzyme kinetics, whereas the expression levels of glucokinase, glucose transporter 2 and insulin receptor were accessed by real-time quantitative RT-PCR. Preliminary results revealed that the Cordyceps-treated groups(the CC, FB and CF groups)showed higher glucose transporter 2 and insulin receptor, than the STZ group. These trends are in accordance to the glycermic status of these groups. For currently unknown reason, expression level of glucose kinase was lower in diabetic rats fed with carcass or fruiting body. Presumably, Cordyceps might exert its anti-hyperglycermic activities through more robust glucose transportation to livers, more effective phosphorylation of glucose, which is the key step for glycolysis, glycogen synthesis, and pentose phosphate pathway, and perhaps more efficient insulin signaling for control glucose homeostasis.
縮 寫 表 VI
中文摘要 - 1 -
Abstract - 2 -
文獻探討 - 3 -
第一節 前言 - 3 -
第二節 冬蟲夏草簡介 - 5 -
一、冬蟲夏草的研究史 - 5 -
二、冬蟲夏草的命名及分類地位 - 5 -
三、冬蟲夏草生活史及特徵 - 8 -
第三節 冬蟲夏草的一般成份 - 10 -
一、一般化學成分 - 10 -
二、冬蟲夏草蛋白質成份 - 10 -
三、冬蟲夏草脂肪酸成份 - 11 -
第三節 冬蟲夏草機能性成份 - 11 -
一、冬蟲夏草的多醣體成份 - 11 -
二、核苷酸 - 12 -
(一)腺苷(adenosine) - 12 -
(二)蟲草素(cordycepin) - 13 -
(三)類似物N6-(2-Hydroxyethyl)adenosine - 14 -
三、固醇類 - 14 -
四、蟲草酸(cordycepic acid or D mannitol) - 16 -
第四節 糖尿病簡介 - 17 -
一、糖尿病定義的修正 - 17 -
二、糖尿病的分類 - 19 -
(一)Type Ⅰ - 19 -
(二)Type Ⅱ - 19 -
(三)特殊類型 - 20 -
(四)妊娠糖尿病 - 20 -
(五)Impaired glucose tolerance - 21 -
三、STZ(streptozotocin)誘發糖尿病鼠之動物模式 - 22 -
(一)基因誘導模式 - 22 -
(二)藥物誘導模式 - 22 -
第五節 葡萄糖代謝及胰島素訊息傳遞 - 25 -
一、葡萄糖激酶(glucokinase) - 25 -
二、葡萄糖轉運蛋白(glucose transporter) - 25 -
三、胰島素的訊息傳遞與葡萄糖運送 - 28 -
(一)胰島素及其受器 - 28 -
(二)胰島素訊息傳遞 - 31 -
(三)葡萄糖生合成的訊息傳遞調控 - 35 -
1、肝醣合成的調控 - 35 -
2、糖質新生作用的調控 - 35 -
3、胰島素阻抗 - 35 -
材料與方法 - 37 -
一、糖尿病大鼠肝臟的取得 - 37 -
二、實驗藥品與儀器 - 38 -
(一)大鼠肝臟分切與製備 - 38 -
(二)大鼠肝臟均質、蛋白質萃取與激酶活性測定 - 38 -
(三)大鼠肝臟組織候選基因表現量之定量 - 40 -
(四)重要實驗儀器 - 49 -
三、實驗方法 - 51 -
(一) 葡萄糖激酶活性測定 - 51 -
1、糖尿病大鼠肝臟組織的分切、均質與粗蛋白質萃取 - 51 -
2、蛋白質標準濃度曲線建立 - 51 -
3、肝臟組織粗蛋白質濃度的定量 - 52 -
4、葡萄糖激酶活性測定 - 52 -
5、葡萄糖激酶活性以比活性方式計算 - 53 -
6、激酶標準品(GK、HK)曲線建立 - 53 -
(二) 肝臟RNA製備與反轉錄 - 54 -
1、RNA製備 - 54 -
2、cDNA製備 - 54 -
3、同步訂量擴增反應 - 55 -
(1) 引子設計(primer design) - 55 -
(2) 核酸同步定量擴增反應條件 - 56 -
四、數據分析 - 58 -
結果與討論 - 64 -
ㄧ、葡萄糖激酶活性測定 - 64 -
二、目標基因 (Glut2、GK、InR) mRNA表現量測定 - 74 -
1、候選基因mRNA表現量測定的前置作業 - 74 -
2、大鼠肝臟GK基因相對表現量測定 - 74 -
3、Glut2 基因相對表現量測定 - 75 -
4、InR基因相對表現量測定 - 76 -
5、各組大鼠肝臟候選基因同步定量產物確認 - 76 -
結 論 - 78 -
參考文獻 - 89 -
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