臺灣博碩士論文加值系統

中文摘要
近年來由於國人飲食習慣西方化，所以膽固醇結石症患者日益普遍，以口服溶石藥物治療膽固醇結石症為患者常用之方法，其風險性低。膽鹽 (BS) 為常用之溶石藥物，目前在臨床上所使用作為溶石藥物的膽鹽有taurochenodeoxycholate (TCDC)、tauroursodeoxycholate (TUDC)，但價格昂貴且帶有副作用。Taurohyodeoxycholate (THDC) 是一種還未為臨床使用的新膽鹽，由豬的膽汁所純化，在來源方面具普遍性，所以為潛力極高的藥物，有必要對此藥物之物化特性作研究，以作為將來成為臨床藥物之依據。本研究對膽鹽臨界微胞濃度 (CMC) 的測定是採用吸光度法與導電度法，在吸光度對於膽鹽濃度之圖形中，偏離比爾定律之點即為CMC，在導電度對於膽鹽濃度圖形中，導電度梯度的改變點即為CMC。在THDC-L溶液系統的K (BS/L) 值研究中，採用25,000 MWCO與12,000~14,000 MWCO兩種孔徑之透析膜作比較，而透析膜外則配製20 mM THDC與40 mM THDC兩種溶液作比較，利用透析膜內外膽鹽濃度差與mixed micelle無法通過透析膜的原理作實驗之設計，透析膜內之溶液則配製THDC/L為80/2、80/4、80/8、80/16、80/24 mM五組作為起始濃度設計，THDC-L溶液系統經5天的震盪放置後會達到系統平衡，再取透析膜內外之溶液作分析，求出系統平衡時之K值。本研究以吸光度法所測定之膽鹽CMC分別為taurocholate (TC)約6 mM，TCDC約2.2 mM，TUDC約2.8 mM，THDC約2.5 mM，以導電度法所測定結果為TC約7 mM，THDC約2.2 mM。平衡透析實驗結果顯示，無論透析膜外所使用之THDC濃度為20 mM或40 mM，都會有個共同現象，當卵磷脂濃度 (CL) 低時，K值較大，隨著CL增加K值會持續降低，最後趨近一定值約1，但使用12,000~14,000 MWCO孔徑之透析膜結果較好，因為卵磷脂經過其膜層所產生之漏出現象較少。

Abstract

Patients of cholesterol gallstones have recently increased in oriental countries since eating habits of oriental people became westernized. To treat this disease, oral administration of drugs in dissolving cholesterol gallstones have been used. Basically, two bile salts (BS), taurochenodeoxycholate (TCDC) and tauroursodeoxycholate (TUDC), are clinically effective; however, both of them are expensive and have side effects. Taurohyodeoxycholate (THDC), a new BS obtained from bile of pigs, has been recently studied to be effective in dissolving cholesterol gallstones in vitro. This result indicates that further investigation of THDC is potential, especially for the study of physicochemical properties of THDC. The critical micelle concentration (CMC) of THDC was determined by using UV-Vis spectrophotometer and conductivity meter. It was showed that a point in a plot of properties vs THDC concentrations which deviated from Beer’s Law was CMC of THDC. Moreover, CMC was estimated by a changing point of conductivity gradient in the plot. To investigate K (BS/lecithin (L)) of mixed micelles in THDC-L solutions by using a dialysis technique, we used two kinds of dialysis membrane which were 25,000 molecular weight cut-off (MWCO) and 12,000~14,000 MWCO. We also used 20 mM THDC and 40 mM THDC initially in receiver side. Moreover, five THDC/L concentrations in donor side, 80/2, 80/4, 80/8, 80/16, and 80/24 mM, were chosen. After 5 days’ of shaking, we analyzed the concentration in donor side and receiver side to determine K values in the equilibrium of THDC-L solutions. The result of CMC of BS by using UV-Vis spectrophotometer is as follows: taurocholate (TC) 6 mM, TCDC 2.2 mM, TUDC 2.8 mM, THDC 2.5 mM. Besides, CMC of TC is 7 mM and THDC is 2.2 mM by using a method of conductivity meter. The experimental outcome of equilibrium dialysis is the same, regardless of 20 mM or 40 mM of THDC used in receiver side. When lecithin concentration (CL) is low, K is large. However, when CL becomes high, K value decreases. Finally, it goes to a constant value (~1). Experimental result shows that dialysis membrane of 12,000~14,000 MWCO is better than that of 25,000 MWCO because of low lecithin leakage.

目錄
中文摘要 ……………………………………………………………...…… i
英文摘要 ………………………………………………………………….. ii
目錄 ………………………………………………………………………. iv
圖目錄 ……………………………………………………………………. vi
表目錄 ……………………………………………………………..……. viii
縮寫對照表 …….………………………………………………….……….x

第一章 緒論

1.1前言 ……………………………………………………………………. 1

第二章 文獻回顧

2.1膽汁分泌 ………………………………………………………………. 7
2.2膽汁中微胞之形成 …………………………………………………… 12

第三章 問題陳述

3.1簡介 ……………………………………………………………..……. 28
3.2平衡透析實驗的缺點 ………………………………………………... 31
3.3複合微胞之組成 …………………………………………………..…. 34

第四章 臨界微胞濃度的測定

4.1簡介 ………………………………………………………………..…. 35
4.2材料與方法 …………………………………………………………... 38
4.3結果與討論 ………………………………………………………..…. 41

第五章 以平衡透析實驗求THDC-L平衡系統之K值

5.1簡介 …………………………………………………………………... 56
5.2材料與方法 ………………………………………………………...… 58
5.3結果與討論 …………………………………………………………... 65

第六章 結論

6.1總論 ……………………………………………………………….….. 85
6.2未來展望 ……..................................................................................…. 87

參考文獻 ……………………………………………………………...…. 88









圖目錄

圖1-1肝膽在消化系統的位置 ….……………………………………….. 5
圖1-2膽固醇結石的生成原因 …………………………………………… 6
圖2-1人類的膽酸 ………..……………………………………………… 19
圖2-2人體膽汁中主要磷脂質的分子結構 …………………………….. 20
圖2-3膽鹽 (TC、TUDC、TCDC、THDC) 的化學結構 ……………... 21
圖2-4人體膽汁中，一級、二級和三級膽鹽合成代謝之化學結構及
位置 ……………………………………………………………… 22
圖2-5膽鹽的腸肝循環 ………………………………………………….. 23
圖2-6 CDC與UDC溶解膽固醇結石機制 ………………………….…. 24
圖2-7複合微胞之類桿狀組成中之主要兩種模型 …………………….. 25
圖2-8平衡透析實驗之裝置圖 ………………………………………….. 26
圖4-1 初功率的輻射被c mole/l吸收物質的溶液吸收及路徑長度
b m所造成的減弱現象 …...………………………………...…. 36
圖4-2 TC臨界微胞濃度之測定 ………………………………………... 43
圖4-3 TCDC臨界微胞濃度之測定 ………………...…………………... 44
圖4-4 TUDC臨界微胞濃度之測定 ………………………...…………... 45
圖4-5 THDC臨界微胞濃度之測定 ……...……………………………... 46
圖4-6以導電度法測定TC之臨界微胞濃度 ………………………...…. 47
圖4-7以導電度法測定THDC之臨界微胞濃度 ……………………….. 48
圖5-1平衡透析實驗中透析膜的作用圖 ……………………………..… 67
圖5-2 THDC溶液之檢量線圖 ……………………………………..…… 68
圖5-3 lecithin溶液之檢量線圖 ……………………………………...….. 69

圖5-4平衡透析實驗之K值與lecithin濃度關係圖 (透析膜孔徑
25,000 MWCO，receiver side為20 mM THDC) ..………….…… 70
圖5-5平衡透析實驗之K值與lecithin濃度關係圖 (透析膜孔徑
25,000 MWCO，receiver side為40 mM THDC) …….………….. 71
圖5-6平衡透析實驗之K值與lecithin濃度關係圖 (透析膜孔徑
12,000~14,000 MWCO，receiver side為40 mM THDC) ….……72
圖5-7 TC溶液之檢量線圖 ……………………………………..……….. 73
圖5-8 TCDC溶液之檢量線圖 ………………………………………….. 74
圖5-9 TUDC溶液之檢量線圖 ……………………………………..…… 75
















表目錄

表2-1人類肝導管之膽汁的組成 ……………………………………….. 27
表4-1 一些常見發色團的吸收特性 ………………………………….... 49
表4-2 TC在pH7.4，NaCl 0.15 M之0.01 M PBS中所測得的
吸光度值 ……………………………………………………...…. 50
表4-3 TCDC在pH7.4，NaCl 0.15 M之0.01 M PBS中所測得的
吸光度值 ………………………………………………………… 51
表4-4 TUDC在pH7.4，NaCl 0.15 M之0.01 M PBS中所測得的
吸光度值 ……………………………………………………...…. 52
表4-5 THDC在pH7.4，NaCl 0.15 M之0.01 M PBS中所測得的
吸光度值 ………………………………………………………… 53
表4-6 TC在pH7.4，NaCl 0.15 M之0.01 M PBS中所測得的
導電度值 ……………………………………………………….... 54
表4-7 THDC在pH7.4，NaCl 0.15 M之0.01 M PBS中所測得的
導電度值 ………………………………………………………… 55
表5-1分析膽鹽時所使用之藥品與藥量 ……………………………….. 76
表5-2不同濃度THDC溶液 (PBS 0.01 M, pH7.4, NaCl 0.15 M) 經
酵素藥品混合後測得之吸光度值 ……………………………… 77
表5-3不同濃度lecithin溶液 (PBS 0.01 M, pH7.4, NaCl 0.15 M) 經
酵素藥品混合後測得之吸光度值 ……………………………… 78
表5-4平衡透析實驗之結果 (透析膜孔徑25,000 MWCO，
receiver side 20 mM THDC) ……....………………...…………… 79
表5-5平衡透析實驗之結果 (透析膜孔徑25,000 MWCO，
receiver side 40 mM THDC) …………...………………………… 80
表5-6平衡透析實驗之結果 (透析膜孔徑12,000~14,000 MWCO，
receiver side 40 mM THDC) ……………...………………...……. 81
表5-7 TC溶液 (PBS 0.01 M, pH7.4, NaCl 0.15 M) 加入酵素混合後
之吸光度測定 ……………………………………………..…….. 82
表5-8 TCDC溶液 (PBS 0.01 M, pH7.4, NaCl 0.15 M) 加入酵素混合後
之吸光度測定 …………………………………………………… 83
表5-9 TUDC溶液 (PBS 0.01 M, pH7.4, NaCl 0.15 M) 加入酵素混合後
之吸光度測定 ………………………………………………...…. 84

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