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研究生:陳韻玨
研究生(外文):Chen Yun-Chueh
論文名稱:中藥純化成份咖啡酸產生降血糖作用之研究
論文名稱(外文):Antihyperglycemic effect of caffeic acid
指導教授:鄭瑞棠鄭瑞棠引用關係
指導教授(外文):Cheng Juei-Tang
學位類別:碩士
校院名稱:國立成功大學
系所名稱:藥理學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:1999
畢業學年度:87
語文別:中文
論文頁數:98
中文關鍵詞:咖啡酸降血糖葡萄糖吸入試驗西方點墨法腺甘酸第一型受體alpha第一型受體
外文關鍵詞:caffeic acidantihyperglycemiaglucose uptakewestern blottingadenosine (subtype1) receptoralpha (subtype1) adrenoceptor
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近年來,罹患糖尿病的患者有日益增加的趨勢,糖尿病也常居國人的十大死因之列。為了協助糖尿病的改善,希望自天然物篩選出有效物質來研發為治療用藥。本項實驗使用兩種糖尿病的老鼠來研究 : 一種是利用 streptozotocin 所誘發的糖尿病老鼠,形成類似糖尿病第一型[胰島素依賴型(insulin-dependent DM, IDDM)]病人的病鼠,另一種則是形成類似糖尿病第二型[非胰島素依賴型( non-insulin-dependent DM, NIDDM)]病人的病鼠。另外,再以普通的老鼠作為正常對照組。Caffeic acid 係純化自蒼耳Xanthium sibiricum patrin (菊科)的果實,一種具有降血糖作用的植物。經由靜脈注射caffeic acid到上述三種老鼠後,在IDDM和NIDDM型的病鼠身上皆可產生依濃度遞增性(dose-dependent)的降血糖作用。可是,caffeic acid 卻不會影響普通老鼠的血糖值。因此,caffeic acid有潛力去發展成為抗高血糖的藥物。接著,藉由葡萄糖挑戰試驗(glucose challenge test),證實這種降低血糖的作用乃經由增加葡萄糖的利用率。在IDDM型老鼠方面,靜脈注射給予adenosine A1受體的拮抗劑 , DPCPX或8-(p-Sulfophenyl)theophylline(8-SPT),或α1受體阻斷劑的prazosin,發現它們皆可抑制caffeic acid的作用。為了解明上述的原因,使用了培養的C2C12細胞(為一種老鼠的肌纖維細胞)來觀察。在離體的脂肪細胞和培養的C2C12細胞,caffeic acid會以濃度遞增性(dose-dependent)的方式,促進[14C] 2-deoxy-glucose進入到細胞內。在培養的C2C12細胞,加入了adenosine A1或α1受體的拮抗劑 ,發現兩者也都在1mM的濃度就可抑制caffeic acid 的作用。同時,利用西方點墨法發現,在培養的C2C12細胞上確實存在著adenosine A1受體。此外, adenosine deaminase的存在也可解消caffeic acid的作用,而dipyridamole則反而可增強caffeic acid的效果 ; 顯示內生性adenosine的參與。而且,利用高壓液相層析法(HPLC)檢測後,發現在培養的C2C12細胞,caffeic acid會以濃度遞增的方式來釋放出adenosine。另外,在受體結合的實驗中,caffeic acid 確實會和[furanyl-5-3H]prazosin 競爭α1受體。因此,caffeic acid可能先作用到α1受體,再藉由內生性adenosine的釋放來產生作用。經由本實驗,找出caffeic acid的降血糖機轉;希望能幫助研發新藥,益於糖尿病患者的治療。

Diabetes mellitus (DM) is one of the common diseases in clinics. In an attempt to develop the new compounds to treat DM, caffeic acid, the compound isolated from the stems of Xanthium sibiricum patrin (Composita), was investigated to know the antihyperglycemic mechanisms. Three types of rats were employed to screen the antihyperglycemic activity of caffeic acid. Wistar rats as normal control were compared with the streptozotocin(STZ)-induced diabetic rats which are similar to the patients with insulin-dependent DM (IDDM) and the rats with insulin-resistance (NIDDM). Caffeic acid produced a dose-dependent hypoglycemic action in both IDDM-type and NIDDM-type rats after an intravenous injection ( i.v.). However, caffeic acid did not lower the plasma glucose in Wistar rats. Thus, caffeic acid has an ability to develop as an antihyperglycemic drug. In the IDDM rats, pretreatment ( i.v.) with selective antagonists of adenosine A1 receptor, 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX) and 8-(p-sulfophenyl)theophylline (8-SPT), diminished the hypoglycemic effect of caffeic acid. Similar result was obtained in STZ-induced diabetic rats receiving i.v. injection of prazosin, the antagonist of α1 receptor. Also, caffeic acid reduced the elevation of plasma glucose level in glucose challenged rats. Both in the isolated adipocytes and the cultured C2C12 myoblasts , a mouse skeletal muscle cell line, caffeic acid increased the uptake of [ 14C ] 2-deoxy-glucose ( 2-DG ) into cells in a dose- dependent manner. In the presence of selective adenosine A1 receptor antagonists, either DPCPX or 8-SPT, the increase of 2-DG uptake by caffeic acid was abolished. Adenosine deaminase at concentration sufficient to metabolize adenosine also attenuated the action of caffeic acid in C2C12 cells. Otherwise, The increase of glucose uptake induced by caffeic acid was enhanced by the coincubation with dipyridamole, the adenosine uptake inhibitor. Involvement of endogenous adenosine can thus be considered. Moreover, western blotting analysis showed the presence of adenosine A1 receptor in C2C12 cells. Secretion of adenosine from cultured C2C12 cell line was increased by caffeic acid in a concentration-dependent manner. Radioligand binding assay showed that caffeic acid can displace the binding of [furanyl-5-3H]prazosin on α1 -adrenoceptor. In conclusion, the antihyperglycemic action of caffeic acid is mainly mediated by an activation of α1 receptor to increase the secretion of endogenesis adenosine.

總目錄 頁數
中文摘要 1
英文摘要 4
縮寫表 7
第一章 緒論 9
第二章 實驗方法及材料 15
第一節 實驗動物 16
第二節 實驗材料 18
第三節 實驗所用之溶液、緩衝液的配製 24
第四節 實驗方法 28
第五節 統計方法 42
第三章 結果 43
第四章 討論 60
第五章 結論 66
參考文獻 68
附圖 75
自述 98

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