臺灣博碩士論文加值系統

先前我們的研究已知在大鼠天麻能減少kianic acid（KA）所誘發的癲癇發作，同時也能減少氧化自由基，我們推測天麻的抗癲癇作用部分來自於它對自由基生成的抑制或清除作用。另外，我們發現天麻也能減少KA誘發的microglia的增殖和活化，以及apoptosis現象。
Activator-protein-1（AP-1）被認為和apoptosis和長期神經細胞可塑性的調節有關，因此本研究的目的是進一步探討AP-1在天麻抗癲癇機轉中所扮演的角色。將15隻雄性的Sprague-Dawley（SD）大鼠分別口服PBS溶液、天麻1.0 克/公斤、0.5克/公斤和Valproic acid （VA）250毫克/公斤一星期，然後從它們的腹腔分別注射12毫克/公斤的KA，經三小時的腦波和肌電圖紀錄，以及行為觀察後，將它們的腦取出，並分離出前額葉皮質和海馬區域的腦組織。最後利用Western blotting分析AP-1上游的活化傳導路徑，包括Extracellular signal-regulated kinase （ERK）、Jun-N-terminal kinase （JNK）和p38 kinase訊息傳導路徑。
結果顯示天麻和VA兩者前治療都能減少KA所誘發的wet dog shakes現象，以及不論是前額葉皮質或hippocampus區域腦組織兩者都經由JNK磷酸化訊息傳導路徑來活化AP-1。
結論是天麻預防性給藥能減少KA 治療大鼠的癲癇發作，但增加AP-1的活性，因此推測天麻可能是利用加速競爭來達到自我保護的作用。另外，天麻活化AP-1是經由MAPK的JNK-P訊息傳導路徑。

Our previous study has known that Gastrodia elata Bl. (GE) can decrease epileptic seizures and oxygen free radicals in Kainic acid (KA)-induced epileptic seizures Sprague-Dawley (SD) rats, suggesting antiepileptic mechanisms of GE results from the suppressive or scavenging effect of free radicals partly. In addition, GE also can inhibit proliferation and activation of microglia, and apoptosis in the rat brain of KA-treated rats.
Several studies finding that activator protein-1 (AP-1) involves neuronal apoptosis and plays a regulatory role in long-term neuronal plasticity. Therefore, the aim of the present study is to investigate the role of AP-1 in antiepileptic mechanism of GE. A total of 15 male SD rats were studied, they were treated with oral administration of PBS solution, GE 1.0g/kg, GE 0.5 g/kg, and valproic acid (VA) 250 mg/kg for one week, respectively, prior to KA 12 mg/kg intra-peritoneal injection. Electroencephalogram (EEG) and electromyogram (EMG) recordings, and behavior were observed for 3 hrs after KA administration, then the rats brain were removed, and cerebral cortex and hippocampus regions of the brain tissue were separated, respectively. Finally, the activating pathways of AP-1, including extracellular signal regulated kinase (ERK), Jun-N-terminal kinase (JNK) and p38 kinase were analyzed by Western blotting. The results indicated that Both GE and VA can decrease wet dog shakes, and AP-1 was activated via mediated a JNK-phosphate (JNK-P) pathway was observed in the frontal cortex and hippocampus region.
In conclusion, GE can decrease epileptic seizures in KA-treated rats, but enhanced AP-1 activity, suggesting that GE possibly accelerated generation of AP-1 to induce a negative feedback for self-protection. In addition, GE is via mediated JNK-P pathway to activate AP-1.
Keywords: Kainic acid，Activator protein-1，Western blotting，Valproic acid，Jun-N-terminal kinase
Our previous study has known that Gastrodia elata Bl. (GE) can decrease epileptic seizures and oxygen free radicals in Kainic acid (KA)-induced epileptic seizures Sprague-Dawley (SD) rats, suggesting antiepileptic mechanisms of GE results from the suppressive or scavenging effect of free radicals partly. In addition, GE also can inhibit proliferation and activation of microglia, and apoptosis in the rat brain of KA-treated rats.
Several studies finding that activator protein-1 (AP-1) involves neuronal apoptosis and plays a regulatory role in long-term neuronal plasticity. Therefore, the aim of the present study is to investigate the role of AP-1 in antiepileptic mechanism of GE. A total of 15 male SD rats were studied, they were treated with oral administration of PBS solution, GE 1.0g/kg, GE 0.5 g/kg, and valproic acid (VA) 250 mg/kg for one week, respectively, prior to KA 12 mg/kg intra-peritoneal injection. Electroencephalogram (EEG) and electromyogram (EMG) recordings, and behavior were observed for 3 hrs after KA administration, then the rats brain were removed, and cerebral cortex and hippocampus regions of the brain tissue were separated, respectively. Finally, the activating pathways of AP-1, including extracellular signal regulated kinase (ERK), Jun-N-terminal kinase (JNK) and p38 kinase were analyzed by Western blotting. The results indicated that Both GE and VA can decrease wet dog shakes, and AP-1 was activated via mediated a JNK-phosphate (JNK-P) pathway was observed in the frontal cortex and hippocampus region.
In conclusion, GE can decrease epileptic seizures in KA-treated rats, but enhanced AP-1 activity, suggesting that GE possibly accelerated generation of AP-1 to induce a negative feedback for self-protection. In addition, GE is via mediated JNK-P pathway to activate AP-1.
Keywords: Kainic acid，Activator protein-1，Western blotting，Valproic acid，Jun-N-terminal kinase
Our previous study has known that Gastrodia elata Bl. (GE) can decrease epileptic seizures and oxygen free radicals in Kainic acid (KA)-induced epileptic seizures Sprague-Dawley (SD) rats, suggesting antiepileptic mechanisms of GE results from the suppressive or scavenging effect of free radicals partly. In addition, GE also can inhibit proliferation and activation of microglia, and apoptosis in the rat brain of KA-treated rats.
Several studies finding that activator protein-1 (AP-1) involves neuronal apoptosis and plays a regulatory role in long-term neuronal plasticity. Therefore, the aim of the present study is to investigate the role of AP-1 in antiepileptic mechanism of GE. A total of 15 male SD rats were studied, they were treated with oral administration of PBS solution, GE 1.0g/kg, GE 0.5 g/kg, and valproic acid (VA) 250 mg/kg for one week, respectively, prior to KA 12 mg/kg intra-peritoneal injection. Electroencephalogram (EEG) and electromyogram (EMG) recordings, and behavior were observed for 3 hrs after KA administration, then the rats brain were removed, and cerebral cortex and hippocampus regions of the brain tissue were separated, respectively. Finally, the activating pathways of AP-1, including extracellular signal regulated kinase (ERK), Jun-N-terminal kinase (JNK) and p38 kinase were analyzed by Western blotting. The results indicated that Both GE and VA can decrease wet dog shakes, and AP-1 was activated via mediated a JNK-phosphate (JNK-P) pathway was observed in the frontal cortex and hippocampus region.
In conclusion, GE can decrease epileptic seizures in KA-treated rats, but enhanced AP-1 activity, suggesting that GE possibly accelerated generation of AP-1 to induce a negative feedback for self-protection. In addition, GE is via mediated JNK-P pathway to activate AP-1.

第一章 前 言 1
第二章 文獻探討
2-1癲癇及其傳統名稱 4
2-2天麻及其歷史沿革 5
2-3天麻現代相關藥理研究 7
2-4 Activator protein 1 的定義 11
2-4-1 Activator protein 1 的活化因子 12
2-4-2 Activator protein 1 的活化方式 14
2-4-3 Activator protein 1對神經生長分化及記憶
產生的活化方式 15
2-4-4 Activator protein 1對腦部炎症反應 16
第三章 材料與方法
3-1實驗動物 17
3-2實驗試劑及儀器（Reagents and Instruments） 17
3-3電極的裝置 19
3-4實驗分組 19
3-5實驗流程
3-5-1行為觀察與腦波記錄 20
3-5-2腦組織的製備 20
3-5-3 Western blotting（西方點墨法） 21
3-5-4西方點墨法實驗步驟 23
3-6統計分析 24
第四章 結 果
4-1 KA誘發SD大鼠行為和腦波的變化 25
4-2天麻在KA誘發癲癇發作大鼠的抗癲癇效用 25
4-3天麻對KA誘發癲癇發作大鼠前額葉皮質AP-1的影響 25
4-4天麻對KA誘發癲癇發作大鼠hippocampus 區域AP-1
的影響 26
第五章 討 論 31
第六章 結 論 33
參考文獻 34
英文摘要 39
謝 辭 41
圖目錄
圖一、Kainic acid腹腔注射誘發痲癇發作腦波和肌電圖的變化 27
圖二、天麻在KA誘發癲癇發作大鼠的抗癲癇作用 28
圖三、天麻對Kainic acid 誘發癲癇發作大腦前額葉皮質的AP-1
的影響 29
圖四、天麻對Kainic acid 誘發癲癇發作大腦的Hippocampus區域
AP-1的影響 30

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