水線草酒精萃取物引發血癌細胞凋亡及促進樹突細胞活化之研究

中國醫藥大學機構典藏 China Medical University Repository, Taiwan > 中醫學院 > 中國藥學暨中藥資源學系暨碩博班 > 博碩士論文 > Item 310903500/46093

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题名:	水線草酒精萃取物引發血癌細胞凋亡及促進樹突細胞活化之研究 Study of human promyelocytic leukemia cell apoptosis and mouse dendritic cell activation induced by ethanolic extract of Hedyotis corymbosa
作者:	林妍甫;Yen-Fu Lin
贡献者:	中國藥學暨中藥資源學系碩士班
关键词:	水線草;樹突細胞;熊果酸;齊墩果酸;血癌細胞;細胞凋亡;Hedyotis corymbosa;HL-60;dendritic cell;ursolic acid;oleanolic acid;apoptosis
日期:	2012-07-26
上传时间:	2012-08-31 16:11:17 (UTC+8)
出版者:	中國醫藥大學
摘要:	水線草為茜草科，是民間常使用的藥用植物之一，可全草入藥，具有清熱解毒之功效。本論文主要探討水線草酒精萃取物及其不同溶劑分層萃取物分別對HL-60細胞之細胞毒殺及老鼠樹突細胞之免疫調節活性的影響。實驗結果顯示，當水線草酒精萃取物的濃度在62.5 μg/mL與其不同溶劑分層萃取物即甲醇及正己烷層濃度分別為11.8 μg/mL及63.4 μg/mL時，經24小時處理，有明顯抑制HL-60細胞50%生長的現象。另外，經由細胞週期的分析結果可知，水線草酒精萃取物亦可使細胞處於sub-G1期的比率增加22.79%，甲醇及正己烷分層萃取物亦分別增加19.01%及21.15%。而更進一步以Annexin V分析方法結果顯示，水線草酒精萃取物使細胞凋亡比率增加至12.6%，而甲醇及正己烷分層萃取物則提升至12.3%及15.7%，並且有劑量及時間依存的情形。而針對免疫調節活性的研究方面，結果顯示水線草酒精萃取物之水分層萃取物，對於樹突細胞之TNF-α的表現量有增加，而水線草酒精萃取物亦可增加IL-6之表現量，顯示其具有促進免疫細胞活化的作用。另外，分別以不同溫度乾燥水線草的全草即常溫乾燥、90℃及50℃烘乾，發現不同乾燥溫度之處理，從HPLC圖譜分析，可見熊果酸、齊墩果酸及兩個未知成分的含量會因不同溫度乾燥處理而改變，然而對於HL-60的抗癌效果並沒有顯著差異。 In Taiwan, Hedyotis corymbosa (Rubiaceae) is a widely used traditional Chinese medicine (TCM), which has been reported in many disease treatments such as cancer, hepatitis and skin disease. The aim of this study is to investigate the anticancer and immuneregulatory activities of ethanol crude extract and its different solvent partitional extracts obtained from H. corymbosa. In this study, we found that ethanol crude extracts of H. corymbosa and its partitional extracts, MeOH, and n-Hexane partitional extracts, exhibited strong inhibition on HL-60 cell growth analysed by WST-1 assay for evaluating cell viability. Besides, above 3 extracts also results in the ratio of standing on sub-G1 stage increasing, DNA fragmentation and apoptosis assay when these extracts were added in HL-60 cells suggested these H. corymbosa extracts induce HL-60 cell death through apoptotic pathway. However, as to the water partitional extracts, it did not show the cell toxicity to HL-60 but enhanced the production of TNF-α and IL-6 in the treated dendritic cell. These results suggest that the water partitional extract of H. corymbosa has immuneregulatory activities. The chemical profiles of H. corymbosa analysed by HPLC was various after drying with different temperature (Room temperature, 50℃ and 90℃). Nevertheless, there are no differences on their apoptotic activities. Two major components, ursolic acid and oleanolic acid, were verified in the extracts of H.corymbosa ethanol crude extract and MeOH partitional extracts using HPLC. In addition, ursolic acid and oleanolic acid may be as the major contents of H. corymbosa on inducing HL-60 apoptosis.
显示于类别:	[中國藥學暨中藥資源學系暨碩博班] 博碩士論文

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