以啟動子轉殖細胞株篩檢增強免疫力之中藥的模式建立

林璧鳳
臺灣大學
日常飲食中有哪些飲食營養因子或食藥兼用之中草藥可能是透過影響生物體內TH-1、 TH-2免疫反應之平衡，至今所知不多。為了探討此問題，本研究試圖以系統性之篩選模式從日常飲食中之食材去找尋具有調節免疫反應之中草藥。研究策略乃利用轉染(transfection)，分別建構能表現TH-1 的細胞激素IFN-γ或TH-2 的細胞激素IL-4或3x-κB-tk啟動子連接報導基因luciferase作為檢測之細胞株 ，例如RAW 264.7巨噬細胞株、EL-4 T淋巴球細胞株等，以作迅速篩選之工具，俾能利於篩選中草藥的抗發炎活性或影響免疫反應之研究，以期建立一個可供天然物、中草藥進行發炎反應研究之不同種類細胞實驗模式。
本計畫分三年進行，第一年，首先利用轉染之細胞株進行細胞培養，當作篩選抗發炎活性中草藥之工具，探討待測樣品對表現IFN-γ、IL-4免疫細胞分子層次及轉錄因子調控的影響。第二年則分別初代腹腔細胞和脾臟細胞以in-vitro給予中草藥材料方式，以及以in-vivo餵食動物方式對腹腔細胞、脾臟細胞分泌發炎性介質之變化，探討轉染之細胞株結果是否與動物模式的結果一致，以便確認此快速篩選模式的有效性，並且了解抑制發炎反應對免疫調節有否影響。第三年以餵食動物中草藥的脾臟細胞與腹腔細胞的免疫指標評估轉染細胞株之應用價值。期望對建立快速篩選既能抗發炎又能增強免疫力之中草藥有正面效果，將來能進一步分析並鑑定功能性成份因子，有助於保健食品之開發。
關鍵字：轉染,細胞激素,IL-4,RAW264.7巨噬細胞株,EL-4 T細胞株,NF-kB

Construction of promoter based immunity screeing system and its application on the study of Chinese Tranditional herbs

Lin, Bi-Fong
National Taiwan University
In order to study the functional effects of numerous Chinese herb medicine on immune responses, a rapid and accurate screening system is needed. The aim of this study is to establish an useful screening system for beneficial effects of dietary factors on immune functions. Cytokines, such as IFN-g, IL-4, NF-kB, have been known to play a very important roles in various physiological events such as the inflammatory response, immunity, differentiation, proliferation and numerous other cellular responses. The ability of dietary factors to modulate these cytokines expression or secretion may be an useful indicators for the possibility to regulate the immune responses. Therefore, this project will be carried out within three years. In the first year, we plan to construct three reporter plasmids containing either IFN-g, IL-4, or 3x-kB-tk promotor gene ligated with luciferase reporter gene respectively. Those reporter plasmids will be used to transfect EL-4 T cell and RAW 264.7 macrophage c
關鍵字：transfection,cytokine,IL-4,Raw264.7,EL-4 T cell,NF-kB