研究生: |
廖助彬 chu-bin Liao |
---|---|
論文名稱: |
N-乙醯基半胱胺酸抑制紫外線照射中國倉鼠卵巢細胞所引發之細胞凋亡 N-acetylcysteine inhibits the UV- induced apoptosis in CHO-K1 cells |
指導教授: |
劉銀樟
yin-chang Liu |
口試委員: | |
學位類別: |
碩士 Master |
系所名稱: |
生命科學暨醫學院 - 生物科技研究所 Biotechnology |
論文出版年: | 2001 |
畢業學年度: | 89 |
語文別: | 中文 |
論文頁數: | 52 |
中文關鍵詞: | 紫外線 、細胞凋亡 、中國倉鼠卵巢細胞 、N-乙醯基半胱胺酸 、氧化自由基 |
外文關鍵詞: | UV, apoptosis, CHO-K1 cells, N-acetylcysteine, reactive oxygen species |
相關次數: | 點閱:4 下載:0 |
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N-acetylcysteine (NAC),是一個具有膜通透性的抗氧化劑,常被用來清除細胞中過多的氧化自由基(reactive oxygen species, ROS),使細胞免於ROS所造成的傷害。先前的研究顯示,紫外線(UV)照射中國倉鼠卵巢細胞(CHO-K1 cells)會促使細胞走向細胞凋亡(apoptosis)。生化分析顯示,CHO-K1 細胞無法表現p21wafl,如果轉殖人類的p21wafl於此細胞中,則可消除UV所造成的細胞凋亡。在本論文研究中,我們發現NAC可以抑制UV所造成的細胞凋亡。雖然NAC可以增加細胞內glutathione(GSH)的量,但是NAC的抑制作用不是透過GSH所造成,因為直接增加細胞內GSH的量並無抑制效果,而且利用DL-buthionine-[S,R]-sulfoximine(BSO)消耗細胞內的GSH並不會加劇UV所造成的細胞凋亡。儘管BSO可以使細胞內ROS的量顯著提升,但ROS的增加並無法加劇UV所造成的細胞凋亡。因此ROS的量與細胞凋亡之間並無直接關係。另外,我們發現利用一種選擇性的cyclin-dependent kinase(CDK)小分子抑制劑-roscovitine,也會抑制UV造成的細胞凋亡。由於NAC可以使細胞生長停滯於G1期,所以我們推論NAC抑制UV所造成的細胞凋亡可能與抑制CDK活性有關,而不是透過抗氧化的機制所造成。
N-acetylcysteine (NAC), a cell membrane permeable antioxidant, is a precursor of glutathione that can remove the excess free radical and protect cells from damage by oxidative stress. Our previously study has indicated that the CHO-K1 cells undergo apoptosis after UV irradiation. The CHO-K1 cells do not express p21wafl, and the UV-induced apoptosis can be inhibited by ectopic expression of human p21wafl gene in the cells. In this study, we found that NAC could protect the cells from the UV-induced apoptosis. Although NAC might increase the intracellular glutathione (GSH) content, the inhibitory effect of NAC on the UV-induced apoptosis was probably not mediated by GSH. Adding GSH into cells failed to produce the similar effect seen with NAC. Moreover, using DL-buthionine-[S,R]-sulfoximine (BSO) to deplete the intracellular GSH content was unable to enhance the UV-induced apoptosis. Despite the large increase of the intracellular ROS content following BSO treatment, the finding that BSO could not promoter the UV-induced apoptosis suggests that the UV-induced apoptosis did not correlate with the intracellular ROS content. In addition, roscovitine, a selective cyclin-dependent kinase (CDK) inhibitor, also inhibited the UV-induced apoptosis in CHO-K1 cells. As NAC was a strong cell cycle inhibitor at G1. The inhibition of NAC upon the UV-induced apoptosis is probably mediated by the modulation of CDK activity as seen with roscovitine.
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