臺灣博碩士論文加值系統

文獻指出為五味子主成份之一之五味子素(Schisandrin B, San B)，具有抗氧化及保護肝臟受損等生物活性及藥理作用。本研究之目的即在探討San B之輻射保護作用與機轉。以流式細胞儀分析發現Chang liver細胞在照射前24小時加入40μM之San B，再以15 Gy放射線照射後，sub G1之比率由放射組的12%降低至5%，顯示照射前加入San B，明顯減少了Chang liver細胞受放射線引發之凋亡。分析其細胞週期之變化發現其G1及S期縮短， G2M期明顯延長，顯示G2關卡增強，將更多之細胞停留在G2M期進行修補，因而增加了細胞之存活率。有關San B對於提升抗氧化之能力方面，於照射前4小時添加40μM之San B，再以放射線照射時，其GSH之濃度明顯比照射組增加。再進一步以西方墨點法分析，在照射前24小時添加40μM之San B，其p53、Bax、caspase 9以及 caspase 3之活性皆明顯下降，而抗凋亡蛋白Bcl-2之活性則維持不變，推論San B之主要藥理乃是經由抑制p53引致細胞之凋亡，因而增加Chang liver細胞之存活。總之，研究顯示San B素可提升細胞之抗氧化能力，抑制放射線照射後引起之細胞凋亡途徑，並增強G2關卡，而達到輻射保護之作用。要之，San B長久使用證實亦無毒性反應，固而San B更具理想之輻射保護劑潛力，其最適有效濃度，則須進一步以動物實驗研究。

Schisandrin B (San B), an active component of Schizandra chinensis Baill, is pharmacologically popular for its bioactivities including anti-oxidative and hepatoprotective effects.
In this present study, we investigated the radioprotective effect of San B and its related mechanism. The effect of San B on the extent of radiation-induced apoptosis was assessed by cell cycle analysis, MTT assay and Western Blot tests. The sub G1 population was found to have increased after irradiation, which was suppressed by pretreatment with San B at 40 mM, by which the apoptotic rate was reduced from 12% to 5% under 15 Gy irradiation (p<0.05). An arrest at G2/M phase was also observed, which apparently could provide a longer time for repairing cell damages resulting in increased cell survival. Pretreatment with 40μM of San B also significantly increased the intracellular glutathione (GSH) levels after irradiation. Furthermore, Western blotting showed that pretreatment with San B significantly down-regulated p53 and Bax, suppressing expression of caspases-9 and caspases-3 after irradiation. However, no effect on the antiapoptotic protein Bcl-2 was found, suggesting that the inhibition of San B on radiation-induced apoptosis is mediated by p53- dependent pathway.
Conclusively, San B could be a promising radioprotectant. Its action mechanism involves antioxidative inhibition of radiation-induced apoptosis. However, cell model did not comply with the cited phenomena revealed by San B. More detail animal study is required for further confirming its radioprotective bioactivities.

中文摘要………………………………….…………………………3
英文摘要…………………………………………………………….6
第一章 研究目的…………………………………………………..8
第二章 文獻回顧…………………………………………….…10
第一節 放射線與細胞凋亡…………………………………….10
1、放射線之治療機轉……………………………………….10
2、細胞凋亡…………………………………………….……12
3、放射線引起之細胞凋亡………………………………….14
3.1 p53調控細胞凋亡途徑………………………..……14
3.2 神經醯胺(ceramide)調控細胞凋亡途徑…….……15
4、p53 基因與凋亡蛋白…………………………………….16
5、細胞凋亡與抗氧化能之關聯性………………………….19
6、放射線對細胞週期之影響……………………………….20
第二節 輻射保護劑……………………………………………….22
1、輻射保護劑……………………………………………….22
2、臨床輻射保護劑作用動力之評析……………………… 23
3、天然植物性輻射保護劑……………………………….…24
第三節 五味子與五味子素…………………………………….25
1、五味子…………………………………………………….25
2、五味子素………………………………………………….26
第三章 實驗方法……………………………………………… 28
3.1五味子萃取物之組成分析……………………….………28
3.2 Chang liver細胞的培養……………………………… 29
3.3.Chang liver生長曲線之測定….………………………29
3.4、五味子素(Schsandrin B)單獨給予之影響………….30
3.5、Chang liver細胞凋亡之流式細胞儀分析…………..30
3.6、Western blot（西方墨點法）……………………… 31
3.7、Glutathione Assay Kit（Sigma）………………… 34
第四章 實驗結果………………………………………………...35
第五章 討論……………………………………………………… 39
第六章 結論……………………………………………………… 44
參考文獻……………………………………………………….….45
圖表……………………………………………………….……….61
附錄…………………………………………………………………70

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