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研究生:陳珉新
研究生(外文):Min-Shin Chen
論文名稱:合成和生物評估第九位置親脂性取代小檗鹼衍生物作為抗癌藥劑之研究
論文名稱(外文):Synthesis of Novel 9-O-/9-N-Lipophilic Substituted Berberine Derivatives and Biological Evaluation as Potential Anticancer Agents
指導教授:吳進益
指導教授(外文):Jin-Yi Wu
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:微生物免疫與生物藥學系研究所
學門:生命科學學門
學類:其他生命科學學類
論文種類:學術論文
畢業學年度:102
語文別:中文
中文關鍵詞:小檗鹼第九位置衍生物HepG2細胞毒性
相關次數:
  • 被引用被引用:1
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肝癌為常見的惡性腫瘤疾病,而在台灣肝癌高居癌症死亡率的第二位,由於肝癌具細胞增生快速的特性進而提高其腫瘤的轉移性與侵犯性。 現今用於治療肝癌病患的常見藥物為doxorubicin,但是會造成病患心臟傷害及肝臟功能衰竭故限制其臨床的使用而導致其五年內生存率低下,因此急切需要可以治療肝癌並提高其預後生存率的治療方式。小檗鹼為一種天然的生物鹼類化合物,主要可於小檗屬類植物的根莖樹皮內萃取而得,且具有多種生物及藥理活性,但是由於其生物鹼的結構導致其親水性較高,而容易被代謝至體外無法有效被小腸吸收。
在本篇研究中,我們利用化學合成方法將小檗鹼的第九位置進行O與N的烷類與萜類官能基修飾,並進行體外對人類癌症細胞株的細胞毒性篩選。而其中 Ber-C15 在人類肝癌細胞株 HepG2 具有最為突出的活性表現,且並不會對正常小鼠胚胎肝細胞 BNL CL.2 產生毒性。在細胞週期與細胞死亡路徑分析試驗中,Ber-C15 能造成肝癌細胞株 HepG2的細胞週期停滯於 S 期且導致細胞凋亡。這些結果顯示Ber-C15 可對於肝癌細胞株 HepG2造成影響,在未來Ber-C15 或許可成為具潛力的抗癌藥物。
目錄
目錄……………………………………………………………………I
中文摘要………………………………………………………………IV
英文摘要………………………………………………………………V
第一章 緒論……………………………………………………………1
第二章 文獻回顧………………………………………………………3
第一節、中藥黃柏之簡介…………………………………………3
第二節、小檗鹼之簡介……………………………………………4
第三節、細胞週期…………………………………………………7
第四節、細胞凋亡…… ……………………………………………9
第三章 研究動機及目的………………………………………………11
第四章 材料與方法……………………………………………………12
第一節、實驗試劑與試藥…………………………………………12
第二節、實驗儀器…………………………………………………15
第三節、黃柏成分萃取方法………………………………………17
第四節、小檗鹼衍生物之合成……………………………………18
第五節、生物活性試驗……………………………………………30
一、細胞株……………………………………………………30
二、繼代培養…………………………………………………30
三、解凍細胞…………………………………………………30
四、冷凍細胞…………………………………………………31
五、計數細胞…………………………………………………31
六、細胞培養試劑之配法……………………………………32
七、細胞存活率………………………………………………33
八、細胞內外化合物含量分析………………………………34
九、細胞週期分析……………………………………………34
十、細胞死亡路徑測定………………………………………36
十一、Hoechst 33258 螢光染色法……………………………37
十二、西方墨點法……………………………………………38
第六節、統計分析…………………………………………………43
第五章 結果與討論……………………………………………………44
第一節、Berberine 衍生物之合成…………………………………44
第二節、探討Berberine 及其衍生物於腫瘤細胞之細胞存活率試驗…………………………………………………………………44
第三節、探討 Berberine 及其衍生物於 HepG2 細胞內外之含量
……………………………………………………………………47
第四節、Berberine 及其衍生物對 HepG2細胞型態之影響
……………………………………………………………………47
第五節、探討 Berberine 及 Ber-C15 對 HepG2 細胞之之細胞週期影響……………………………………………………………48
第六節、探討 Berberine 及 Ber-C15 對 HepG2 細胞之死亡路徑
……………………………………………………………………49
第七節、探討 Berberine 及 Ber-C15 對細胞週期相關蛋白之影響
……………………………………………………………………50
第六章 結論……………………………………………………………51
第七章 圖表……………………………………………………………53
參考文獻………………………………………………………………62
附錄……………………………………………………………………73
參考文獻
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