臺灣博碩士論文加值系統

本實驗之目的在探討不同的 CAPE 類似物 EC、BrCAPE，對 PBMC ( peripheral blood mononuclear cell ) 細胞是否有細胞毒性及對其分泌細胞激素的影響。實驗共分成二個部分：一、將 PBMC 細胞培養在合成物中24小時及48小時，以 MTT ( microculture tetrazolium assay )分析細胞活性；二、將 PBMC 細胞培養在合成物中24小時及48小時，取其上清液，分別測定其 IL-4，IL-6，Human GM-CSF，Human Soluble fas 濃度上的變化。結果顯示，這三種合成物並無細胞毒性存在；且對 IL-4 及 Soluble fas 分泌的影響較大，也較一致，故可推論 CAPE 類似物可能具有相同的生物活性。經由 IL-4 及 Soluble fas 免疫調節的影響，細胞產生自戕作用及細胞免疫的上升，對於癌細胞是否能有效抑制，可再深入探討其作用機制，使之對人類有更大貢獻。

The purpose of this experiment is to study the cell toxicity and the effect of PBMC excreted cytokines by the CAPE-like analogues, 3-(3,4-dihydroxy -phenyl) acrylic acid ethyl ester (EC), 3-(4-bromophenyl) acrylic acid phenethyl ester (BrCAPE). In this study, experiment was divided into two parts. (1) Incubated PBMC in the analogues for 24 and 48 hours. Using MTT (microculture tetrazolium assay) to analyze the cell activity. (2) Incubated PBMC in the analogues for 24 and 48 hours. Analyzing the change of the four cytokines, IL-4, IL-6, Human GM-CSF and Human Soluble fas by taking the suspender. The results of the study showed that all the analogues was no cell toxicity and had heavily change in the secretion of IL-4 and Human Soluble fas. So, we could conclude that the CAPE-like analogues had the same biologic activity. At the influence of the IL-4 and Human Soluble fas, the cell immunity and apoptosis went up. Whether the CAPE-like analogues could inhibit cancel cell or not, it would be another topic to study to improve the human being life.

目錄
第一章 緒言‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥1
第二章 文獻回顧‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥2
第一節 PBMC 細胞‥‥‥‥‥‥‥‥‥‥‥‥‥‥2
第二節 細胞激素‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥3
一、 白間素‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥3
二、 株落刺激因子‥‥‥‥‥‥‥‥‥‥‥‥‥3
三、 IL-4‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥3
四、 IL-6‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥4
五、 其他細胞激素‥‥‥‥‥‥‥‥‥‥‥‥‥‥4
六、 輔助型T細胞‥‥‥‥‥‥‥‥‥‥‥‥‥‥4
第三節 蜂膠類的化合物‥‥‥‥‥‥‥‥‥‥‥‥5
第三章 材料與方法‥‥‥‥‥‥‥‥‥‥‥‥‥‥6
第一節 實驗材料‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥6
一、 測試儀器‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥6
二、 生物活性之相關試劑‥‥‥‥‥‥‥‥‥‥‥6
三、 細胞激素之相關試劑‥‥‥‥‥‥‥‥‥‥‥7
四、CAPE, EC, BrCAPE 的製備‥‥‥‥‥‥‥‥‥9
第二節 實驗方法與步驟‥‥‥‥‥‥‥‥‥‥‥12
一、 PBMC細胞之培養‥‥‥‥‥‥‥‥‥‥‥12
二、 細胞活性分析‥‥‥‥‥‥‥‥‥‥‥‥‥13
三、 Cytokines的測定‥‥‥‥‥‥‥‥‥‥‥‥13
四、 標準之製備‥‥‥‥‥‥‥‥‥‥‥‥‥‥14
第四章 結果‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥16
第五章 討論‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥22
第六章 結論‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥27
參考文獻‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥28
表目錄
表 1. Cytotoxicity of CAPE，EC，BrCAPE in PBMC cells evaluated by MTT assay (treated 24 hours)‥‥‥‥‥‥‥‥‥38
表 2. Cytotoxicity of CAPE，EC，BrCAPE in PBMC cells evaluated by MTT assay (treated 48 hours)‥‥‥‥‥‥‥‥‥39
表 3. Interleukin-4 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-4 set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥40
表 4 Interleukin-6 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-6 set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥41
表 5. Human GM-CSF secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human GM-CSF set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥42
表 6. Human Soluble Fas secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human Soluble Fas set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥43
表 7. Interleukin-4 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-4 set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥44
表 8 Interleukin-6 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-6 set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥45
表 9. Human GM-CSF secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human GM-CSF set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥46
表 10. Human Soluble Fas secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human Soluble Fas set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥47
表 11. 細胞激素的比較(24及48小時)‥‥‥‥‥‥23
圖目錄
圖 1. Cytotoxicity of CAPE，EC，BrCAPE in PBMC cells evaluated by MTT assay (treated 24 hours)‥‥‥‥‥‥‥48
圖 2. Cytotoxicity of CAPE，EC，BrCAPE in PBMC cells evaluated by MTT assay (treated 48 hours)‥‥‥‥‥‥‥49
圖 3. Interleukin-4 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-4 set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥50
圖 4 Interleukin-6 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-6 set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥51
圖 5. Human GM-CSF secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human GM-CSF set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥52
圖 6. Human Soluble Fas secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human Soluble Fas set assay (treated 24 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥53
圖 7. Interleukin-4 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-4 set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥54
圖 8 Interleukin-6 secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human IL-6 set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥55
圖 9. Human GM-CSF secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human GM-CSF set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥56
圖 10. Human Soluble Fas secretion by PBMC cells stimulated by CAPE，EC，BrCAPE evaluated by Human Soluble Fas set assay (treated 48 hours)‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥57
圖 11. 標準組‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥58
圖 12. 對照組(24小時)‥‥‥‥‥‥‥‥‥‥‥‥59
圖 13. 對照組(48小時)‥‥‥‥‥‥‥‥‥‥‥‥60
圖 14. CAPE，EC，BrCAPE結構圖‥‥‥‥‥‥‥61

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