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QR Code 本論文永久網址: 複製永久網址

Twitter 研究生: 施睿琥

研究生(外文): Jui-Hu Shih

論文名稱: 利用正子造影於大鼠模式探討白藜蘆醇抗MDMA神經毒性之保護作用 論文名稱(外文): Exploring the Neuroprotective Effects of Resveratrol against MDMA-induced Neurotoxicity in Rats Using Positron Emission Tomography 指導教授: 李宜勳 指導教授(外文): I-Hsun Li 口試委員: 馬國興、吳錦楨、王信二、顏若芳、李宜勳 口試委員(外文): Kuo-Hsing Ma、Chin-Chen Wu、Hsin-Ell Wang、Ruoh-Fang Yen、I-Hsun Li 口試日期: 2015-12-29 學位類別: 博士 校院名稱: 國防醫學院

系所名稱: 醫學科學研究所 學門: 醫藥衛生學門 學類: 醫學學類 論文出版年: 2016 畢業學年度: 104 語文別: 中文 論文頁數: 76 中文關鍵詞: 白藜蘆醇、搖頭丸、血清素轉運體、正子造影、佔有率 外文關鍵詞: resveratrol、MDMA、serotonin transporter、positron emission tomography、occupancy 相關次數: 被引用:0

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下載:48 書目收藏:0 由於3,4-methylenedioxymethamphetamine (MDMA) 對人類的心理與生理有嚴重影響，MDMA之濫用已引起全球越來越多的關注。MDMA濫用者往往伴隨著持久性血清素神經毒性，這與腦部血清素轉運體 (serotonin transporter；SERT) 密度降低和憂鬱症兩者息息相關。白藜蘆醇 (resveratrol；RSV) 是一種天然多酚之植物性防禦素，已被證實具有抗憂鬱和神經保護之作用。然而，白藜蘆醇之生理作用標靶及其對MDMA保護作用仍未研究透徹。因此，本研究利用正子造影結合血清素轉運體之放射性配體N,N-dimethyl-2-(2-amino-4-[18F]fluorophenylthio)benzylamine (4-[18F]-ADAM)，探討白藜蘆醇與MDMA對於血清素轉運體之結合能力；另外，我們亦研究白藜蘆醇是否能具有防止MDMA急性與長期性不良反應之保護作用。實驗結果發現，白藜蘆醇對於活體大鼠各腦區之血清素轉運體具有劑量依賴之結合潛力。當同時給予大鼠皮下注射10 mg/kg MDMA與腹腔注射20 mg/kg 白藜蘆醇，每日兩次連續4天，可顯著降低MDMA引起血漿皮質固醇濃度之急性上升。此外，透過4-[18F]-ADAM正子造影證實白藜蘆醇於給藥後兩週，能有效防止MDMA所誘導之血清素轉運體可用率下降；由強迫游泳試驗亦可發現白藜蘆醇能充分改善MDMA大鼠之類憂鬱行為。綜合上述，白藜蘆醇確實是一個具有抗憂鬱及對抗MDMA引發神經生物及行為變化之潛力藥物。

3,4-methylenedioxymethamphetamine (MDMA) is a widely misused drug that has been reported to evoke acute weight loss and hypothalamic-pituitary-adrenal axis activation, but also induces long-term psychophysiological changes in rodents, promoting depressive-like behavior and reducing the density of brain serotonin transporters (SERT). Resveratrol (RSV) is a natural polyphenolic phytoalexin and known for its antioxidant, antidepressant, and neuroprotective effects. However, biological targets of RSV as well as its neuroprotective effects against MDMA are still poorly understood. In this study, we examined binding potency of RSV and MDMA to SERT using small-animal positron emission tomography (PET) with the SERT radioligand, N,N-dimethyl-2-(2-amino-4-[18F]fluorophenylthio)benzylamine (4-[18F]-ADAM) and investigated the protection of RSV against the acute and long-term adverse effects of MDMA. RSV was found to exhibit binding potentials to SERT in vivo in a dose-dependent manner with variation among brain regions. To assess the protection by RSV against the acute and long-term adverse effects of MDMA, we injected animals with RSV (20 mg/kg, i.p.) and MDMA (10 mg/kg, s.c.) twice daily for 4 consecutive days. Small-animal PET imaging data showed that RSV protected against the MDMA-induced decrease in SERT availability in the midbrain and the thalamus 2 weeks following the co-treatment. RSV also prevented the MDMA-evoked increase in immobility in the forced swim test. Remarkably, RSV inhibited the MDMA-induced elevation in plasma corticosterone. Together, these findings suggest that RSV is not only a potential antidepressant, but that it also confers protection against the neurobiological and behavioral effects of MDMA.

正 文 目 錄

正文目錄------------------------------------------------------------------------------------Ⅰ

表目錄-------------------------------------------------------------------------------------Ⅲ

圖目錄-------------------------------------------------------------------------------------Ⅳ

中文摘要-----------------------------------------------------------------------------------Ⅵ

英文摘要-----------------------------------------------------------------------------------Ⅶ

第一章、緒言--------------------------------------------------------------------------------1

第二章、材料與方法--------------------------------------------------------------------------12

第一節、化學試劑----------------------------------------------------------------------------12

第二節、實驗動物----------------------------------------------------------------------------12

第三節、放射藥物與動物正子造影---------------------------------------------------------------13

第四節、實驗一：利用4-[18F]-ADAM正子造影測量白藜蘆醇或MDMA之血清素轉運體佔有率-------------------14

第五節、實驗二：評估白藜蘆醇對於MDMA之急性期與長期作用之影響------------------------------------17

第六節、採血步驟----------------------------------------------------------------------------21

第七節、LC tandem mass分析血漿皮質固醇濃度試驗------------------------------------------------21

第八節、糖水偏好試驗------------------------------------------------------------------------22

第九節、強迫游泳試驗-------------------------------------------------------------------------23

第十節、統計分析----------------------------------------------------------------------------23

第三章、結果--------------------------------------------------------------------------------25

第一節、利用4-[18F]-ADAM正子造影評估白藜蘆醇與血清素轉運體之結合能力-----------------------------25

第二節、白藜蘆醇減緩MDMA於停藥後期間之體重減輕作用----------------------------------------------32

第三節、白藜蘆醇抑制MDMA引發血漿皮質固醇濃度---------------------------------------------------37

第四節、白藜蘆醇減少MDMA大鼠之靜止不動與增加游泳行為--------------------------------------------39

第五節、白藜蘆醇具有對抗MDMA引起血清素轉運體喪失與類憂鬱行為之保護作用----------------------------41

第四章、討論--------------------------------------------------------------------------------47

第五章、結論--------------------------------------------------------------------------------56

第六章、參考文獻----------------------------------------------------------------------------57





表 目 錄

表1. 動物與人體等效劑量換算公式--------------------------------------------------------------19

表2. 白藜蘆醇與MDMA之血清素轉運體佔有率 (%)---------------------------------------------------27

表3. 強迫游泳試驗與血清素轉運體可用率之相關矩陣------------------------------------------------44





圖 目 錄

圖1. MDMA化學結構式--------------------------------------------------------------------------2

圖2. MDMA 於血清素神經元末稍與突觸之藥理作用機轉------------------------------------------------3

圖3. 4-[18F]-ADAM化學結構式------------------------------------------------------------------6

圖4. resveratrol化學結構式-------------------------------------------------------------------8

圖5. 神經突觸體製備流程圖--------------------------------------------------------------------10

圖6. 白藜蘆醇於活體外神經突觸體具有抑制攝取 [3H]5-HT之作用--------------------------------------11

圖7. 4-[18F]-ADAM正子造影評估白藜蘆醇與MDMA之血清素轉運體佔有率之實驗流程------------------------16

圖8. 白藜蘆醇對MDMA急性與長期影響之實驗流程圖---------------------------------------------------20

圖9. 大鼠靜脈注射不同劑量白藜蘆醇 (溶媒、10、20、40、80 mg/kg) 與皮下注射10 mg/kg MDMA之正子造影--28

圖10. 靜脈注射不同劑量白藜蘆醇與血清素轉運體佔有率之關係-----------------------------------------29

圖11. 不同劑量白藜蘆醇之血清素轉運體加權平均佔有率----------------------------------------------31

圖12. MDMA與白藜蘆醇對體重影響之經時變化-------------------------------------------------------34

圖13. MDMA與白藜蘆醇於停藥期對體重之影響-------------------------------------------------------35

圖14. 白藜蘆醇對糖水偏好試驗之影響-------------------------------------------------------------36

圖15. 白藜蘆醇抑制MDMA引發血漿皮質固醇濃度------------------------------------------------------38

圖16. 白藜蘆醇對MDMA大鼠游泳行為之影響---------------------------------------------------------40

圖17. 白藜蘆醇提供大鼠對抗MDMA引發血清素轉運體可用率降低之保護作用--------------------------------43

圖18. 靜止不動秒數與各腦區專一性攝取率散佈圖----------------------------------------------------45

圖19. 正常大鼠各腦區血清素轉運體之經時變化------------------------------------------------------50

圖20. 白藜蘆醇與MDMA之共同藥理機轉-------------------------------------------------------------55





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