摘要
以 NG108-15 細胞為模式，探討以三種不同機制增加胞內鈣的藥物對細胞死亡之影響胞內鈣庫，鈣幫浦抑制劑 thapsigargin 不但會造成細胞死亡，也會促使神經突生長，前者經由 caspase 之活化，後者則是經由 PLA2 之活化二者均是鈣獨立的。其他的鈣邦浦抑制劑如 BHQ 及 CPA 也能促使神經突生長。質子通透劑，FCCP，能瓦解粒線體膜電位，使粒線體無法吸取鈣，造成胞內鈣上升，FCCP 也會活化 caspase 造成細胞死亡，但是此過程密切依賴胞外鈣。Phospholipase C 連結的胞外激素，bradykinin，在生理狀況下，能提升胞內鈣濃度，對細胞形態及細胞死亡均無影響。TG 雖然與 FCCP 一樣能同時活化 caspase ，但在引起細胞死亡上較慢，TG 也許也能促使一保護因子的活化，進而減緩細胞死亡的進行。本論文並未確認此保護因子為何物，但可確知不是 NO，cAMP， arachidonic acid，GDNF 或 NT-3。在沒有胞外鈣存在下，FCCP 及 TG 也都能促使細胞死亡。我們的結果顯示，在NG108-15 細胞，細胞死亡可依循鈣依賴的及鈣非依賴的二種途徑進行。

Abstract
Using neuroblastoma x glioma hybrid NG108-15 cells as the model ayatem , we characterized the effect of three different Ca2+ elevating agents on the cell death and cell growth . Intracellular Ca2+ ATPase (sarcoplasmic and endoplasmic reticulum Ca2+ ATPase, SERCA) inhibitor, thapsigargin (TG), not only activated phospholipase A2, but also stimulated caspase. the former caused neurite outgrowth, while the latter resulted in cell death. Both events were Ca2+ independent. Other SERCA inhibitors, including BHQ and CPA, displayed similar effect as TG in terms of the neurite outgrowth. The proton ionophore, FCCP, dissipated the proton gradient of mitochondria which in turn caused the collapse of the mitochondria membrane potential and the leak of trapped Ca2+. FCCP aslo activated caspase and caused cell death. However, it is extracellular Ca2+ dependent. Bradykinin, the phospholipase C coupled hormone, increased the cytosolic Ca2+ level under physiological condition and had no effect on morphology and the process of cell death. The activation of caspase evoked by TG or FCCP occurred simultaneously. However, the process of the cell death induced by TG was slower than that by FCCP. It is possible that TG may activate a cellular protective factor which delay the process of cell death. We have not identified this protective factor yet, however, it is not any of NO, cAMP, arachidonic acid, GDNF and NT-3. In the absence of extracellular Ca2+, TG- or FCCP-induced cell death was still observed. In conclusion, our results indicate that in NG108-15 cells, the process of cell death occurs via either Ca2+ dependent or Ca2+ independent pathway.

正文目錄
頁次
正文目錄...............................................................................................................I
圖次目錄...............................................................................................................II
縮寫表..................................................................................................................III
中文摘要..............................................................................................................IV
英文摘要................................................................................. ............................V
緒論...................................................................................................................... 1
實驗材料..............................................................................................................8
實驗方法.............................................................................................................10
結果............................................................................. ........................................15
討論..................................................................................................................... 20
參考文獻.............................................................................................................38
圖表目錄
頁
圖一：在 NG108-15 細胞，FCCP、TG 及 BK 所引起之
胞內鈣上升。····································24
圖二：FCCP，TG，BK，CPA 及 BHQ 對 NG-108-15 細
胞型態之影響。······························· ····26
圖三：FCCP、TG 及 BK 引起的 arachidonic acid 釋放，
cAMP 及 NO 的產生。·································27
圖四Arachidonic acid及 20碳脂肪酸對細胞型態之影響。····· ···············28
圖五：各式抑制劑對TG引起的神經突長之影響。·······················29
圖六：PLA2 抑制劑對TG引起的神經突長之影響。······················30
圖七：Arachidonic acid 代謝途徑抑制劑對 TG 造成神經
突長之影響。····································31
圖八：FCCP，TG 及 BK 引起之 Caspase 活化。·········· ·············32
圖九：FCCP，TG 及 BK 引起的細胞死亡。·········· ···············33
圖十：FCCP，TG 及 BK 引起的細胞死亡。········ ·················34
圖十一：FCCP，TG 及 BK 所引起的 GDNF 分泌。······················35
圖十二：FCCP，TG 及 BK 所引起的 NT-3 的分泌。······· ··············36
圖十三：S-1-P 及 Arachidonic acid 對 FCCP 引起的細胞
死亡之影響。································· ··37

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