臺灣博碩士論文加值系統

TRPV1 channel為一個鈣離子通透的通道，將capsaicin刺激人類骨肉瘤細胞G292後可以觀測到細胞中鈣離子上升，且此鈣離子的來源為細胞中的內質網鈣庫，並可以發現細胞中鈣離子上升會有濃度依存的現象。利用capsaicin耗空內質網鈣庫後，可造成G292細胞死亡，且死亡是經由粒線體內在路徑cytochrome c的釋放及caspase-3的活化。此外capsaicin亦會活化ERK與誘發細胞自噬(autophagy)的產生。在分別加入細胞自噬的誘發劑rapamycin與細胞自噬的抑制劑3-MA時，capsaicin所造成的ERK的磷酸化分別會增加與減少，表示細胞自噬(autophagy)位於ERK的上游並具有調控ERK磷酸化的功能。當加入pan-caspase的抑制劑ZVAD-fmk時可以降低capsaicin造成的細胞死亡，但在同時加入pan-caspase的抑制劑ZVAD-fmk與細胞自噬的抑制劑3-MA，則無法抑制capsaicin所誘發的死亡，表示capsaicin所誘使的caspase-independent細胞死亡可以被細胞自噬與ERK所抑制，相反地，同時加入pan-caspase的抑制劑ZVAD-fmk與細胞自噬的誘發劑rapamycin，則capsaicin所誘發的死亡會降低，表示細胞自噬對於capsaicin所誘發的細胞死亡具有保護的作用。總結上述結果，在G292細胞中，capsaicin會誘使鈣庫鈣離子耗空並經由粒線體內在路徑活化caspase-dependent的死亡路徑，同時亦會活化細胞自噬與ERK來拮抗caspase-independent的死亡路徑。

TRPV1 is a calcium permeable channel. Stimulating human osteosarcoma G292 by capsaicin will elevate intracellular calcium, and the calcium is released from calcium pool, endoplasmic reticulum. We also find that the amount of intracellular calcium released depends on the capsaicin concentration. Using capsaicin to deplete intracellular calcium pool will cause cell death in G292 cell and the death is through intrinsic mitochondrial pathway by cytochrome c release and activation of caspase-3. Besides, capsaicin will also activate P44/42 MAPK and induce autophagy. Treating G292 cell with autophagy inducer rapamycin and inhibitor 3-MA will increase and decrease P44/42 MAPK phosphorylation respectively. These indicate the autophagy is at the upstream of the P44/42 MAPK and can regulate P44/42 MAPK activity. Treating G292 cell with pan-caspase inhibitor ZVAD-fmk can decrease cell death caused by capsaicin, but treating cell with pan-caspase inhibitor ZVAD-fmk and autophagy inhibitor 3-MA at the same time can not decrease cell death caused by capsaicin. These suggest that caspase-independent cell death induced by capsaicin can not be blocked by autophagy and P44/42 MAPK. In contract, treating cell with pan-caspase inhibitor ZVAD-fmk and autophagy inducer rapamycin at the same time will decrease cell death caused by capsaicin. These indicate autophagy can protect cell from death induced by capsaicin. All of the above, capsaicin will deplete intracellular calcium pool, endoplasmic reticulum, and induce intrinsic mitochondrial death pathway through activation of caspase, and also activate autophagy and P44/42 MAPK to block caspase-independent death pathway.

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正文目錄…………………………...…...……………………………………I
圖文目錄……………………………......………………………………...….II
縮寫表………………………………......…………………………………....III
中文摘要……………………………...…...…………………………...…….IV
英文摘要………………………………...…………………………...............V
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第一章、緒論…………………………...……………………………………1
第二章、實驗方法與材料…………...…...………………………………….6
第三章、結果……………………………...…………………………………13
第四章、討論…………………………......………………………………….19
第五章、結論…………………………......………………………………….23
第六章、參考文獻………………………...…………………………………31

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