1.探討Pentylenetetrazol所引起的抽搐對未成熟老鼠腦中c-fos , c-jun , bax , bcl-2 , cystatin B及p53基因表現之影響2.探討低濃度Arachidonic acid對大白鼠腎上腺髓質嗜鉻腫瘤細胞神經發育之影響

The effect of pentylenetetrazol-induced seizure on the expression of c-fos , c-jun , bax , bcl-2 , cystatin B and p53 gene in the brain of developing rats2.The effect of low concentration arachidonic acid on neuronal differentiation in rat Pheochromocyto

黃麗穎(Li-Ying Huang)

指導教授 ： 葉健全

臺北醫學大學/醫學院/醫學科學研究所/碩士(2002年)

摘要

Pentylenetetrazol（PTZ）是一種痙攣劑，它會阻斷r-aminobutyric acid接受體（GABAA receptor）的神經性傳導抑制作用，而引起成熟與未成熟動物的抽搐行為。過去已經知道PTZ作用在成鼠身上時，會導致腦部細胞死亡，推測其原因可能與apoptosis的產生有相關。所以在本實驗中，我們在未成熟的老鼠身上施打PTZ後，以Northern blotting的方法來定量在cortex, hippocampus, striatum與cerebellum腦區中c-fos, c-jun, bax, bcl-2, cystatin B與p53這些與apoptosis相關基因的表現，並且用N-methyl-D-aspartate（NMDA）接受體的非競爭性拮抗劑MK-801來阻斷PTZ所引起之抽搐行為與其相關之基因表現。由實驗的結果發現，PTZ會使7、14及30天大的老鼠產生明顯的generalized seizure，但若先在老鼠身上施打MK-801，則可以完全抑制seizure的產生。而Northern blotting的結果顯示，PTZ會使所有腦區的c-fos表現增加，且其表現與年齡及腦區有密切的關係；而施打MK-801也可以有效抑制住因PTZ所產生c-fos。這樣的結果顯示，PTZ會引發immediate early gene的表現，且需要NMDA receptor的活化。至於c-fos的表現是否意謂著會走向apoptosis的路徑，還有待更進一步的了解。

關鍵字

Pentylenetetrazol ； r-aminobutyric acid接受體 ； N-methyl-D-aspartate ； NMDA ； 接受體 ； seizure ； 細胞凋亡 ； MK-801

並列摘要

Pentylenetetrazole (PTZ), a convulsant agent, induces seizure in both mature and developing animals by blocking the r-aminobutyric acid type (GABAA) receptor-mediated inhibitory neurotransmission. The PTZ-induced seizure has been found to produce brain injury in adult rats, partly attributed to the generation of apoptosis. In the present study, I determined whether PTZ could induce apoptosis in the developing rats by quantifying the expression of apoptosis associated genes, namely, c-fos, c-jun, bax, bcl-2, cystatin B and p53 gene in the cortex, hippocampus, striatum and cerebellum of rats received single injection of PTZ using Northern blotting assay. I also determined whether PTZ-induced seizure and associated gene expression could be blocked by addition of a noncompetitive antagonist of N-methyl-D-aspartate (NMDA) receptor, the MK-801. The result showed that PTZ induced generalized profound seizure on rats with age of 7, 14, and 30 days. Pre-injection of MK-801 completely abolished the seizure. Northern blotting assay showed that injection of PTZ only induced apparent expression of c-fos in all examined brain regions but with an age- and region-specific manner. Injection of MK-801 abolished PTZ-induced expression of c-fos. This result indicates that PTZ is able to trigger the immediate early gene expression and this action requires activation of the NMDA receptor. Whether the expression of c-fos alone means the ongoing of apoptosis or other effect unrelated to apoptosis remains to be elucidate.

並列關鍵字

PTZ ； GABAA receptor ； NMDA receptor ； seizure ； apoptosis ； MK-801

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