甲基培尼皮質類固醇(MPD)在臨床上是常見治療慢性發炎的藥物，而茶為日常普遍被飲用的飲料，此兩者皆被研究出對於骨質生長有一定的影響。本實驗探討大鼠體內甲基培尼皮質類固醇與沒食子酸脂化兒茶素(EGCG)在藥物動力學的交互作用，及合併甲基培尼皮質類固醇與茶葉萃取物對大鼠骨頭的藥效學交互作用。
使用高效液相層析－電化學系統分析大鼠血漿中EGCG的含量，在實驗組合併給予腹腔注射MPD (5 mg/kg)及口服EGCG(1 g/kg)的大鼠體內，MPD增加EGCG時間－濃度曲線下面積(AUC)約34% (p≦0.05)，但合併給予MPD及EGCG對於血漿中最大濃度(Cmax)及身體清除率(clearance)對控制組單獨口服EGCG (1 g/kg)則沒有顯著性差異，推測甲基培尼皮質類固醇可透過與腸道P-醣蛋白的結合而促使EGCG進入血液循環中。
利用4週給藥模式給予大鼠腹腔注射MPD (5 mg/kg)單獨給予或合併口服不同的茶葉萃取物(400 mg/kg)，使用微電腦斷層掃描儀偵測大鼠股骨骨小樑結構總骨量(BV/TV，%)的變化，發現單獨給予MPD、MPD合併茶葉粗萃物及MPD合併高含量EGCG茶葉萃取物皆可輕微增加骨小樑的總骨量BV/TV (21.64 ± 6.8% : 23.04 ± 7.1% : 24.39 ± 4.3% : 27.73 ± 1.8%)，尤其MPD合併去咖啡因且高含量EGCG茶葉萃取物有顯著性增加骨小樑總骨量的表現(32.24 ± 6.1%；p = 0.032)，顯示短期單獨使用MPD或MPD合併茶的飲用皆對骨質密度有正向的效益。

Methylprednisolone (MPD), an anti-inflammation drug, is frequently used for long-term treatment of chronic inflammation disorders. Tea is a popular beverage worldwide. Both of tea and MPD have been reported to affect bone mineral density to a certain extent. This study was aim to investigate the effect of MPD on the pharmacokinetics of epigallocatechin-3-gallate (EGCG, a tea polyphenol). In addition, the pharmacodynamic interaction of MPD and tea extracts with respect to bone mineral density was also discussed.
Experimental Sprague–Dawley (SD) rats were treated with MPD (5 mg/kg, i.p.) immediately after EGCG (1 g/kg, oral) administration, whereas the control group received EGCG (1 g/kg, oral) only. The plasma EGCG was measured by high performance liquid chromatography with electrochemical detection. The pharmacokinetic data indicate that the area under the plasma concentration–time curves (AUC) of EGCG was increased by 34% for the group with MPD treatment (p≦0.05). As regards EGCG maximum concentration and clearance, there were no significant differences between these two groups. Mehtylprednisolone might bide with intestine P-glycoprotein and promoted EGCG into blood circulation.
For the pharmacodynamic experiments, SD rats were treated with MPD (5 mg/kg, i.p.) in the absence or presence of different tea extracts (400 mg/kg, oral) for four weeks. Micro-CT scanning was used to measure various skeletal microstructural parameters. The results showed that MPD along or coadministrated with tea extract, EGCG enriched tea only slightly increased the BV/TV value (sham: 21.64 ± 6.8%; MPD: 23.04 ± 7.1%; MPD/crude extract: 24.39 ± 4.3%; MPD/EGCG enriched extracts: 27.73 ± 1.8%). However, the BV/TV value increased significantly in the group treated with MPD/decaffeine and EGCG enriched tea extract (32.24 ± 6.1%; p= 0.032). Together, our findings suggested that short-term usage of MPD along or with tea might have benefit to bone density.

Abstract……………………………………………………………..….VIII
中文摘要………………………………………………………………..…X
一、緒論……………………………………………………………...……1
1-1 Methylprednisolone簡介與其相關研究……………………………1
1-1-1研究背景………………………………………………….......1
1-1-2藥理作用與骨骼生長的相關研究……………………….......2
1-2 Epigallocatechin-3-gallate(EGCG)簡介與其相關研究……………..3
1-2-1 研究背景……………………………………………………..3
1-2-2 藥物基本特性與藥理作用…………………………………..5
1-2-3 相關藥物動力學之研究……………………………………..8
1-3 P-醣蛋白……………………………………………………………..9
1-3-1 P-醣蛋白背景介紹…………………………………………....9
1-3-2 P-醣蛋白對藥物動力學的影響...............................................10
1-3-3 Methylprednisolone對於P-醣蛋白的調控與影響.................10
1-4 藥物動力學…………………………………………………………...13
1-4-1 藥物動力學概論……………………………………………..13
1-4-2 藥物動力學的定義…………………………………………..13
1-4-3 藥物動力學參數....................14
二、研究動機與目的.......................14
三、實驗材料與方法.......................17
3-1 實驗材料………………………………………………………….....17
3-1-1 試藥與試劑…………………………………………………17
3-1-2 儀器設備與實驗材料………………………………………18
3-1-3實驗動物…………………………………………………….19
3-1-4 試藥的配製…………………………………………………19
3-1-5 應用軟體……………………………………………………20
3-2甲基培尼類固醇(Methylprednisolone, MPD)與沒食子酸酯化兒茶素(EGCG)藥物動力學實驗.................21
3-2-1 動物手術步驟………………………………………...……21
3-2-2 檢體採集時間與採血方式……………………………...…21
3-2-3 血液檢體處理步驟……………………………………..…22
3-2-4 高效液相層析-電化學(HPLC-EC)之分析條件..................23
3-2-5 血液檢量線之配製……………………………………..…23
3-2-6 層析系統之精確性(precision)及準確性(accuracy)……....24
3-2-7 樣品處理回收率之測定………………………………..…25
3-3甲基培尼類固醇(Methylprednisolone, MPD)與沒食子酸酯化兒茶素(EGCG)藥效學實驗……………………………………………………26
3-3-1 茶葉萃取物成分之分析方法…………………………..…28
3-3-2 實驗動物給藥方法……………………………………..…29
3-3-3 實驗動物犧牲、灌流與取骨…………………………..…30
3-3-4 微電腦斷層( Micro Computer Tomatography)偵測骨小樑分佈情形…………………………………………………………... 31
四、實驗結果…………………………………………………...………31
4-1甲基培尼類固醇(Methylprednisolone, MPD)與沒食子酸酯化兒茶素(EGCG)藥物動力學之分析結果………………………………………31
4-1-1 血液中的藥物層析分析………………………………….31
4-1-2 大鼠血液檢品分析校正曲線之確認………………….…31
4-1-3 血液中藥物的回收率………………………………….…32
4-1-4 大鼠血漿中沒食子酸脂化兒茶素之經時變化……… …32
4-1-5 血漿中沒食子酸脂化兒茶素之藥物動力學的參數…… 33
4-2甲基培尼類固醇(Methylprednisolone, MPD)與沒食子酸酯化兒茶素(EGCG)藥效學之分析結果……………………………………………33
4-2-1茶葉萃取的分析結果………………………………… …33
4-2-2 微電腦斷層掃描之股骨斷層圖………………………… 34
4-2-3 微電腦斷層掃瞄之骨小樑參數……………………… …35
五、實驗討論…………..………………………………………………37
5-1 甲基培尼類固醇(Methylprednisolone, MPD)與沒食子酸酯化兒茶素(EGCG)藥物動力學研究之討論………………………………………37
5-1-1 實驗動物採樣之選擇……………………………….……37
5-1-2 電化學分析EGCG的方法討論.........38
5-1-3 Methylprednisolone對血漿中EGCG濃度之藥物動力學的影響……………………………………………………………...…38
5-2 甲基培尼類固醇(Methylprednisolone, MPD)與沒食子酸酯化兒茶素(EGCG)藥效學研究之討論……………………………………….….. 39
5-2-1 實驗動物模式的選擇………………………………….…39
5-2-2 茶葉萃取物對大鼠骨密度的影響……………………….40
六、結論………………………………………………………..………42
參考文獻…………………………………………………………….….43

附表目錄
表一、甲基培尼類固醇(MPD)與沒食子酸酯化兒茶素(EGCG)藥效學實驗之給藥組別……………………………………………………………..28
表二、測定血漿中EGCG經液-液萃取的回收率……………………53
表三、利用高效液相層析系統測量EGCG在血漿中同日與間日的準確度accuracy(%)與精確度precision(R.S.D.)………………………….…....54
表四、經由口服給予大鼠EGCG 1g/kg後，計算藥物於血漿中的藥物動力學參數………………………………………………………………...55
表五、茶葉萃取物中兒茶素類之含量…………………………...……56
表六、大鼠連續給藥四週後，偵測大鼠股骨骨小樑在型態學參數的變化
……………………………………………………………………………57

附圖目錄
圖一、甲基培尼皮質類固醇(methylprednisolone)的化學結構式….....1
圖二、沒食子酸脂化兒茶素 (EGCG) 的化學結構式……………......6
圖三、血漿的高效液相層析圖…………………………...…………...58
圖四、口服EGCG 1 g/kg後，於大鼠血漿中所偵測到EGCG的濃度-時間曲線關係圖……….....................................59
圖五、茶葉萃取物的高效液相層析圖…………………………..........60
圖六、大鼠給藥四週後股骨骨小樑結構分佈的情形………………..…61
圖七、大鼠給藥四週後股骨骨小樑總骨量BV/TV(%)的表現量……………62
圖八、大鼠用藥四週後股骨骨小樑數目Tb．N (1/mm)的表現量…………………63
圖九、大鼠用藥四週後股骨骨小樑分離情形Tb．Sp (mm)...........64

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