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研究生:陳翠霞
研究生(外文):CUEI-SIA, CHEN
論文名稱:在鼠腦皮質細胞中, Levetiracetam保護由麩胺酸引起的細胞傷害
論文名稱(外文):The Protect effect of the Anti-epileptic Drug Levetiracetam on Glutamate-induced Cell Damage in Cortical Neuron
指導教授:闕小輝
指導教授(外文):Sheau-Huei Chueh
口試委員:王正康劉佩珊
口試委員(外文):Jehng-Kang, WangPei-Shan,Liu
口試日期:2013-05-24
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:31
中文關鍵詞:麩胺酸
外文關鍵詞:Glutamate
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本研究主要利用鼠大腦皮層神經細胞作為材料,探討抗癲癇藥物Levetiracetam對於Glutamate促使細胞傷害的影響。由出生24小時內的SD老鼠取得腦皮質神經細胞,貼盤培養再有poly-D-lysine的coverslip上。加入10µM Ara-C培養至少三天後,加入Levetiracetam(Lev) 300µM培養並觀察對於其是有保護細胞不受Glutamate的傷害。結果顯示,在有預先給予Levetiracetam的組別在細胞的死亡率、ROS的產生均低於Glutamate的組別。以Glutamate刺激引導胞內鈣濃度的變化也是在有預先給予Levetiracetam的組低於control組別。但當加入鈣螯合劑EGTA後,細胞被Glutamate傷害的現象就消失,不論細胞是否預先給予Levetiracetam死亡率、細胞數均未有明顯變化,說明Glutamate造成的死亡是透過鈣依賴的方式,而Levetiracetam保護的方式是因可以降低胞外鈣內流入胞內。
In this studying we used rat cortical neuron as the Material to understand the effect of anti-epileptic drug Levetiracetam (Lev) on glutamate-induced cell damage. We got cortical neuron from 24 hours postnatal SD rat’s cortex, cultured with 10µM Ara-C in coverslip containing poly-D-lysine at last 3 days(DIV 3).
We pre-treated Levetiracetam 300µM, than stimulated cells with glutamate to see if Levetiracetam could protect cell. The result exhibited Levetiracetam reduced glutamate-induced cell damage in cell death and the generation of ROS. We also found that pre-treated Levetiracetam can lower the glutamate-induced calcium influx. Then we added the calcium chelator EGTA to blocked calcium. We observed that after adding EGTA, the effect of glutamate-induced cell damage is disappeared. Thus, our concluded that Levetiracetam can protect cortical neuron to against glutamate-induced cell damage through the calcium- depend pathway.

正文目錄
緒論………………………………………………………..……………….……1
第一節、介紹神經細胞……………………………………...…………………..1
第二節、過氧化氫………………………….………………..…………………..5
第三節、抗癲癇藥物Levetiracetam…………………………………………....6
第四節、研究動機………………………….…………………………………..7
第二章材料與方法………………………….…………………………………..8
第一節、實驗材料………………………….…………………………………..8
壹、實驗動物………………………….…………………………………….…..8
貳、實驗藥品………………………….…………………………………….…..8
叁、實驗儀器………………………….…………………………………….…..8
第二節 實驗方法…………………….…………….……………………….…..9
壹、大腦皮質初級細胞培養……….……………...……………………….…..9
貳、免疫細胞化學染色法……….…………………………………….………10
叁、活性氧物質之測定……….…………………………………….………….10
肆、細胞死亡測定……….…………………………………….……………….11
伍、細胞內鈣離子測定.……………………………………….……………….11
第三章實驗結果.…………………………………………..….……………….13

第一節細胞型態與定性…………………………………..….……………….13
第二節、藥物Levetiracetam對glutamate所造成的細胞傷害之保護作用...13
第三節、藥物Levetiracetam對H2O2所造成的細胞傷害之保護作用……....14
第四節、藥物Levetiracetam對鈣離子內流(Ca2+ influx)的影響…………..15
第五節、藥物Levetiracetam對細胞內ROS產生的影響………………….15
第四章、討論…………………………………………………………………..17
第五章、結論…………………………………………………………………..18
第六章、參考文獻……………………………………………………………...30

第六章 參考文獻
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