苻苓在中國傳統醫藥上，廣泛地被使用來治療水腫及腹瀉。近代有更深入的研究，顯示苻苓也有抗發炎及抗腫瘤的療效；然而，關於這些功能的作用機制仍未被揭示。在本研究中，我們藉由觀察苻苓對結腸上皮的離子運輸之調節效用，嘗試尋找可以解釋它的上述功能之機制。本研究利用Ussing chamber的技術，來測定苻苓對結腸腸膜上皮膜電位及短路電流之影響。結果顯示苻苓施於黏膜側時，可明顯增加短路電流。Amiloride可以抑制由苻苓所引起的短路電流改變，顯示苻苓是作用於上皮對amiloride敏感的鈉離子通道；另一方面，苻苓施用於漿膜側時，所測得之短路電流沒有增加太多甚至減少，顯示它對於鈉鉀幫浦的影響甚微。我們的數據顯示氯離子阻斷劑bumetanide及chlorothiazide也可以抑制由苻苓所引起的短路電流改變，顯示苻苓所引起的短路電流有部分可能是經由氯離子流動所提供。我們的結論是:苻苓可以增強結腸上皮對鈉離子的吸收、並刺激氯離子分泌。

PCWE has been used widely in oriental traditional medicine in treating edema and diarrhea. Recent studies have shown that PCWE may also have anti-tumor and anti-inflammation acts. However, few studies have been conducted to reveal the mechanisms of these effects. In the present study, we tried to elucidate the mechanisms by investigating the effects on PCWE on the regulation of ion transport across the epithelial membranes of colon, which is also useful in explaining the anti-diarrhea and anti-edema effects. Alternation in membrane potential and short-circuit current (Isc) were examined using the Ussing chamber technique. Our results showed that PCWE decreased Isc upon application to the apical side. Amiloride inhibited this Isc induced by PCWE indicating that PCWE acted on amiloride-sensitive sodium channel of the epithelium. However, when PCWE was applied to the serosal side, the Isc was not changed, indicating a minimal influence of this substance on ATP-driving Na+/K+ counter transporters. Our data also showed that the Isc decreased by PCWE could be inhibited by bumetanide and chlorothiazide (Cl‾ channel inhibitors). We therefore concluded that PCWE could both enhance sodium transport and stimulate the secretion of Cl‾ in colon epithelium.

目錄
頁次
中文摘要------------------------------------------------ 5.
英文摘要------------------------------------------------ 6.
文獻回顧
一、 苻苓成份與作用------------------------------------------------ 7-9.
二、 中藥利尿藥物與化學利尿劑作用機轉之關聯------------------------ 9-11.
三、 幫浦滲漏假說與細胞水腫之間的關係------------------------------ 11-12.
四、 細胞膜上的離子幫浦-------------------------------------------- 12-15.
五、 生物性電位理論-雙膜假說--------------------------------------- 15-16.
六、 上皮細胞容量的改變與鈉氯離子運輸有關--------------------------- 16-18.
七、 哺乳類的結腸上皮---------------------------------------------- 19.
八、 哺乳類腸道上皮電解質吸收及分泌的細胞模式---------------------- 20.
九、 結腸上皮的水分子運輸------------------------------------------ 21.
十、 偵測鈉離子流的藥物(Amiloride)與上皮鈉離子通道(ENaC)----------- 22-25.
十一、 胞外ATP於上皮細胞的訊號傳導------------------------------ 25-26.
十二、 Purinergic receptor對上皮運輸的調節----------------------- 27-29.
研究目標------------------------------------------------- 30.
實驗方法及材料
一、實驗器材:
電壓嵌制系統及記錄儀器---------------------------------------- 31.
二、組織預備:----------------------------------------------------- 31.
三、使用溶液:----------------------------------------------------- 32.
四、Ussing system
電極製作------------------------------------------------------ 33.
測量短路電流-------------------------------------------------- 34.
五、藥物影響短路電流及其電導值計算方式---------------------------- 35.
六、統計方法------------------------------------------------------ 36.
七、實驗流程------------------------------------------------------ 36-37.


結果
A. PCWE作用於皮膜兩側對其短路電流及電導值之影響-------------------- 38.
B. Amiloride對PCWE造成之短路電流及電導值影響---------------------- 38-39.
C. Chlorothiazide對PCWE造成之短路電流及電導值影響----------------- 39-40.
D. Bumetanide對PCWE造成之短路電流及電導值影響--------------------- 40-41.
E. PCWE、Chlorothiazide與Bumetanide依序對皮膜短路電流的影響------- 41.
F. PCWE、ATP與Amiloride對皮膜短路電流的交互影響------------------- 42-43.
G. PCWE加入漿膜側對其短路電流及電導值之影響------------------------ 43-44.
討論--------------------------------------------------------------- 45-50.
結論--------------------------------------------------------------- 51.
參考文獻---------------------------------------------------------- 52-59.
圖表目錄
頁次

表一.實驗中各種藥物對於皮膜作用後的短路電流(Isc)、電導值(Gt)及電流變
化百分比(△Isc)列表。---------------------------------------- 60.
圖一.苻苓(PCWE)主要成分之化學結構---------------------------------- 61.
圖二.苻苓(PCWE)作用於黏膜側及漿膜側之短路電流圖-------------------- 62.
圖三.苻苓(PCWE)單獨作用於黏膜側，並分別與ATP、鈉及氯離子阻斷劑進行作
用之短路電流統計圖------------------------------------------- 63.
圖四.苻苓(PCWE)單獨作用於黏膜側，並分別與ATP、鈉及氯離子阻斷劑進行作
用之短路電流圖------------------------------------------------ 64.
圖五.Amiloride單獨作用於黏膜側，並分別與苻苓(PCWE)、ATP進行作用之短
路電流統計圖-------------------------------------------------- 65.
圖六. a. ATP先處理黏膜側後與Amiloride、苻苓(PCWE)進行作用
b. Chlorothiazide先處理黏膜側後與苻苓(PCWE)進行作用
c. Bumetanide先處理漿膜側後，再與苻苓(PCWE)進行作用，
之短路電流統計圖---------------------------------------------- 66.
圖七.Amiloride單獨作用於黏膜側，並分別與苻苓(PCWE)、ATP進行作用之短
路電流圖------------------------------------------------------ 67.
圖八.ATP單獨作用，並分別與苻苓(PCWE)、Amiloride 進行作用之短路電流圖-------------------------------------------------------------- 68.
圖九.Chlorothiazide及Bemetanide個別單獨作用於黏膜側及漿膜側，再與苻
苓(PCWE)進行作用之短路電流圖---------------------------------- 69.
圖十.苻苓(PCWE)、ATP單獨作用於漿膜側，及苻苓(PCWE)後再加入ATP以進行
作用之短路電流統計圖------------------------------------------ 70.

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