Quantitative Detection of ECP Level of STA-1 Treated HL-60 Cell by Using real-time RT-PCR

應用real-time RT-PCR檢測人類白血病細胞經STA-1培養後ECP之表現

張東迪(Tung-Ti Chang) ； 許清祥(Ching-Hsiang Hsu) ； 黃介辰(Chien-Chen Huang)

《臺灣中醫醫學雜誌》 5卷1期 (2006/06) Pp. 41-59

https://doi.org/10.30032/TJCM.200606.0005

摘要

麥門冬湯與六味地黃丸之合方萃取物，STA-l能改善氣喘病患之臨床症狀及肺功能。本研究探討STA-l之抗發炎分子機轉。因此我們選擇ECP為標的，作為探討STA-l對此ECP表現之調控實驗，並因此藉以建立一細胞模型作為將來評估其它中藥調控ECP表現與發炎機轉之分子模型。本實驗將中藥，丁酸與人類白血病細胞HL-60 clone 15 cells一起培養2小時與24小時後，以相對定量real-time RT-PCR檢測ECP mRNA表現量，並以house keeping gene，β-actin，作內部定量控制(internal control)。實驗結果顯示，經由2小時與24小時的中藥、丁酸與HL-60 clone 15 cells培養後實驗組對ECP表現量之向上調控與對照組比較有統計上差異。本研究顯示以HL-60 clone 15 cells為材料，可作為監測ECP基因表現之很好模型。

關鍵字

即時反轉錄聚合酶鏈鎖反應 ； 中藥萃取物 ； 嗜伊紅性陽離子蛋白 ； 人類白血病細胞

並列摘要

Though we have showed that the extracts of STA-l, a traditional Chinese medicine (TCM) formulae composed of Mai-Men-Dong-Tang (MMDT) and Lui-Wei-Di-Huang-Wan (LWDHW), could provide improvement in lung function and relieve asthma symptoms without adverse effects, molecular markers were needed to monitor the effect of STA-l on cell level. The gene of eosinophil cationic protein (ECP) was choose in this study to establish a model for investigating the effect of STA-l on ECP expression. The real-time RT-PCR was used to quantify the expression level of ECP mRNA levels in HL-60 clone 15 cells and a house keeping gene, β-actin, was used as the control for quantification. A significant expression of ECP by both STA-l and butyric acid treatment were showed during 24 hours culturing. The results suggest that expression level of ECP within HL-60 host cell may be a good candidate model to monitor the effect of Traditional Chinese Medicine, STA-l.

並列關鍵字

RT-PCR ； STA-1 ； ECP ； HL-60