臺灣博碩士論文加值系統

傳統中醫認為癲癇發作有部分是由於肝系統中陰陽間的不平衡，導致肝風內動的結果。因此用平肝息風的方法來治療癲癇發作。天麻是中藥的一種，它被認為具有平肝息風的作用，自古以來就被用來治療癲癇發作。我們先前的研究已知天麻能減少kainic acid（KA）誘發大白鼠的癲癇發作，天麻的這個作用與氧化自由基生成的抑制和清除有關。另外，我們發現天麻也能抑制microglia的活化，因此本研究的目的是探討天麻的抗癲癇作用與interleukin-1β(IL-1β)、tumor necrosis factor-α(TNF-α)，以及一氧化氮（nitric oxide，NO）的關係。於Sprague-Dawley(SD)大白鼠腹腔注射KA(12mg/kg)誘發癲癇發作，以及前治療天麻1.0 g/kg 、0.5 g/kg，以及valproic acid (VA) 250 mg/kg四天來觀察天麻的效用，整個實驗過程都施行行為觀察和腦波與肌電圖記錄。行為觀察和腦波、肌電圖記錄是從KA注射前的15 min到注射後的90 min為止，我們計算wet dog shakes的發作次數，並從大白鼠的左心室採血3.0 cc測量IL-1β、TNF-α以及NO的濃度。最後取腦，分別測定前額葉與海馬區域的IL-1β、TNF-α濃度。結果天麻1.0g/kg和VA 250mg/kg能減少KA誘發的wet dog shakes次數。KA能增加週邊血液的 IL-1β和NO濃度，但這些增加能被天麻1.0 g/kg和VA 250 mg/kg所抑制。另外，KA能增加前額葉IL-1β和TNF-α以及海馬區域TNF-α濃度，這些增加也都能被天麻1.0 g/kg和VA 250 mg/kg所抑制。
結論是天麻能抑制KA誘發癲癇發作大鼠細胞因子中IL-1β、TNF-α以及NO的產生，可能是天麻經由抑制microglia的活化，而減少IL-1β、TNF-α對microglia的強化，使iNOS活性降低，導致NO和自由基產生減少，而抑制癲癇發作。

In Traditional Chinese Medicine, the etiology of epileptic seizure is considered that it is due to the imbalance between Yi and Yang in the liver system, resulting in liver wind stirring internally. Therefore, the method of calm the liver to stop the wind is used to treat epileptic seizure. Gastrodia elata Bl.(GE) is one of Chinese herbs, and it is considered that has the action of calm the liver to stop the wind, and is used to treat epileptic seizure for a long period. Our previous studies have known that GE can reduce kainic acid (KA)-induced epileptic seizures in rats, and this effect of GE has closely relationship the suppression or scavenging action of oxygen free radicals. In addition, we also find that GE can inhibit the activation of microglia, therefore, the purpose of the present study to investigate the relationship between anticonvulsive effects of GE and interleukin-1 (IL-1β), tumor necrosis factor-α(TNF-α), and nitric oxide (NO). Animal model of epileptic seizures was performed by KA (12 mg/kg) i.p., and pretreatment with GE 1.0 g/kg, 0.5 g/kg and valproic acid (VA) was done for 4 days, respectively, to observe the effect of GE, the behaviors of the rats and electroencephalogram (EEG) and electromyogram (EMG) were recorded throughout the experimental course. The behavioral observation, EEG and EMG recordings were from 15 min before to 90 min after KA injection. The total counts of wet dog shakes were calculated, and blood of 3 cc was taken from heart for the measurement of IL-1β, TNF-αand NO. Finally, the brains of the rat were taken out for the measurement of IL-1βand TNF-αin the frontal cortex and hippocampus region. The results indicated that both pretreatment of GE 1.0 g/kg and VA 250 mg/kg could reduce the counts of wet dog shakes. The increase of IL-1βand NO in the peripheral blood of KA-treated rats, and these increase can be inhibited by pretreatment with GE 1.0 g/kg or VA 250 mg/kg. In addition, the levels of IL-1βand TNF-αincreased in the frontal cortex, and TNF-αin the hippocampus region in the KA-treated rats, whereas these increase also can be inhibited by pretreatment of GE 1.0 g/kg and VA 250 mg/kg.
In conclusion, GE inhibit the generation of IL-1β, TNF-αand NO in the KA-induced epileptic seizures rats, possibly GE mediate via inhibition of microglia activation resulting in IL-1βor TNF-αdecrease to the potent microglia to synthesize inducible nitric synthase (iNOS). Thus the generation of NO and free radicals decreased results in the inhibition of epileptic seizure.

指導教授推薦書…………………………………………………
口試委員會審定書………………………………………………
誌謝……………………………………………………………… iii
中文摘要………………………………………………………… iv
英文摘要………………………………………………………… vi
第一章 背景………………………………………………… 1
第二章 文獻探討…………………………………………… 3
2.1 西方醫學對癲癇的認識與探討……………………… 3
2.2 歷代中國醫學對癲癇的認識與探討………………… 7
2.3 中藥天麻的認識與探討……………………………… 8
2.4 細胞因子（cytokine）中IL-1β及TNF-α與NO跟癲癇
關係的探討…………………………………………… 17
第三章 材料與方法………………………………………… 21
3.1 實驗動物……………………………………………… 21
3.2 試劑及儀器…………………………………………… 21
3.3 實驗流程……………………………………………… 22
3.3.1電極的裝置…………………………………… 22
3.3.2 Pilot study………………………………… 23
3.3.3 實驗分組……………………………………… 23
3.3.4 行為觀察與腦波記錄………………………… 24
3.3.5 腦組織的萃取………………………………… 24
3.3.6腦組織中細胞因子IL-1β和TNF-α以及血液中細胞因
子IL-1β和TNF-α跟NO的測定………… 25
3.3.7 統計分析………………………………………… 26
第四章 結果 …………………………………………………… 27
4.1 KA誘發SD大鼠行為和腦波的變化…………………… 27
4.2天麻對KA誘發癲癇發作大白鼠的抗癲癇作用………… 29
4.3天麻對KA誘發癲癇發作大白鼠周邊血液IL-1β、
TNF-α以及NO的影響…………………………………… 31
4.4天麻對KA誘發癲癇發作大白鼠前額葉及海馬回IL-1β
和TNF-α的影響……………………………………… 33
第五章 討論……………………………………………………… 36
第六章 結論……………………………………………………… 38
參考文獻 …………………………………………………… 39

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