臺灣博碩士論文加值系統

內生性enkephalin系統是否可以調控創傷後壓力症候群（post-traumatic stress disorder，PTSD）目前沒有清楚確定的研究結果。而本研究的目的是以C57BL/6J（wild-type，WT）小鼠和enkephalins基因剔除（preproenkephalin knock-out，ppENK KO）小鼠來探討缺乏內生性enkephalins與PTSD的關係，並藉此建立一個PTSD的動物模式。 此動物模式主要以強迫性電擊刺激作為創傷事件的來源，並使用僵直行為反應、開放性區域檢測（OFT）、十字型高架迷津檢測（EPM）、明暗趨避檢測（LDT），以及強迫性游泳檢測（FST）來觀測老鼠的反應，並利用免疫組織化學反應來檢測c-fos的反應。結果顯示，經過電擊的老鼠比沒有電擊的老鼠表現出更多焦慮和憂鬱的反應，並且ppENK KO小鼠對於經過電擊所引發的反應比WT強烈，而經過電擊的ppENK KO小鼠之c-fos反應可在與PTSD相關的腦區看出有明顯增加的現象，尤其是Cg/RS、CA1和CA2等腦區。根據上述結果可知：（1）ppENK KO小鼠在接受重大壓力後，比WT更容易產生焦慮和憂鬱的反應。（2）本研究結果釐清了內生性enkephalin系統與PTSD的關係。

The functional role of enkephalins in the trauma-induced stress processing is not clear. The aim of the present study was to investigate the involvement of the endogenous enkephalin system in animal model of post-traumatic stress disorder (PTSD). All mice encountered either a 3 mA or no inescapable footshock for 10 seconds, and sixteen wide type (WT) mice and sixteen preproenkephalin knockout mice were assigned into four groups: Shock-WT, Shock-KO, No Shock-WT, and No Shock-KO. Subsequently, situational reminders were given 1 min for three trials and mice’s freezing behaviors were assessed on the day 2, 7, and 13. On the day 14, all mice were randomly tested for open filed test (OFT), elevated plus-maze (EPM), light/dark avoidance test (LDT), and forced swim test (FST). Mice with footshock showed more anxious and depressive responses on the OFT, EPM, LDT, and FST than mice without footshock. Moreover, the magnitude of freezing behavior for KO mice is significantly greater than those of WT mice. The c-fos expression has a significant enhancement in the brain regions, such as Cg/RS, CA1, and CA2, of KO mice experienced a footshock. In summary, mice with the enkephalin gene knockout are susceptible to chronic stress-induced anxiety and depression. Our experimental results have provided clearly evidences of the involvement of endogenous enkephalin system in the PTSD symptoms.

目次
第一章 緒論 ................................................. 1
第一節 引言 ........................................................ 1
第二節 PTSD的動物模式 ............................................. 2
第三節 內生性鴉片類系統與PTSD的關係 .............................. 4
第四節 研究目的與假設 .............................................. 6
第二章 材料與方法 ........................................... 8
第一節 受試者 ...................................................... 8
第二節 聚合酵素鏈鎖反應 ............................................ 8
第三節 實驗儀器與程序 .............................................. 9
一、強迫性電擊刺激 ............................................. 9
二、情境的提示 ................................................ 11
三、僵直行為反應 .............................................. 11
四、十字型高架迷津檢測 ........................................ 11
五、明暗趨避檢測 .............................................. 12
六、開放性區域檢測 ............................................ 13
七、強迫性游泳檢測 ............................................ 13
第四節 免疫組織化學反應 ........................................... 15
一、 灌流手術與組織固定 ........................................ 15
二、 免疫組織化學反應 .......................................... 15
三、脫水與封片 ................................................. 16
第五節 實驗流程 ................................................... 16
第六節 統計分析 ................................................... 19
第三章 結果 ................................................ 20
第一節 聚合酵素鏈鎖反應 ........................................... 20
第二節 僵直行為反應 ............................................... 22
第三節 動物行為檢測 ............................................... 24
一、十字型高架迷津檢測 ........................................ 24
二、明暗趨避檢測 .............................................. 26
三、開放性區域檢測 ............................................ 28
四、強迫性游泳檢測 ............................................ 30
第四節 免疫組織化學反應 ........................................... 33
第四章 討論 ................................................ 40
參考文獻 ................................................... 44
附錄 ....................................................... 58
附錄一 DSM-IV-TR關於PTSD的診斷標準 .......................... 58
附錄二 縮寫表 .................................................. 62

圖次暨表次
Figure 1 實驗用的電擊箱 ............................................... 10
Figure 2 四種不同的動物行為檢測 ....................................... 14
Figure 3 實驗流程圖 ................................................... 18
Figure 4 聚合酵素鏈鎖反應的結果 ....................................... 21
Figure 5 在情境提示中的僵直行為表現結果 ............................... 23
Figure 6 十字型高架迷津檢測的結果 ..................................... 25
Figure 7 明暗趨避檢測的結果 ........................................... 27
Figure 8 開放性區域檢測的結果 ......................................... 29
Figure 9 強迫性游泳檢測的結果 ......................................... 32
Figure 10 沒有經過電擊的老鼠之免疫組織化學反應的結果 .................. 36
Figure 11 經過電擊的老鼠之免疫組織化學反應的結果 ...................... 37

Table 1 野生型小鼠與ppENK基因剔除小鼠的c-fos反應數量之平均值 ......... 39

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