聚木醣酶基因转型乳酸球菌之益生效果探讨.DOC

甘草次酸對HepG2解毒酵素誘發作用之探討 Phase II enzyme induction effect of glycyrrhetic acid in HepG2 cell lines 研究生：徐偉真 Wei-Jen Shiu 指導教授：黃卓治 Tzou-Chi Huang 【摘要】 傳統上，中醫廣泛使用甘草作為調合諸藥、抗潰瘍、抗發炎、解毒、鎮咳祛痰等治療，許多研究證實甘草中的有效藥理成分甘草次酸(Glycyrrhetic acid, GA)在解毒、抗炎、增强免疫調節中扮演重要的角色。肝臟是身體最重要的解毒器官，利用產生解毒酵素將有毒物質進行代謝且排除體外，以減低對身體的傷害。因此，本實驗為研究甘草次酸在人類肝癌細胞(HepG2)對解毒酵素誘發之影響。由實驗結果得知甘草次酸可以誘發Phase II解毒酵素，包括麩胱甘肽轉移酶(GST)、麩氨酸半胱氨酸接合酶(GCLc)等，在處理12小時時有最高的表現量，其中GCLc為合成麩胱甘肽 (glutathione, GSH) 的重要酵素，而GSH的表現量與GST的活性在甘草次酸的誘發下於12小時有顯著增加。由流式細胞儀分析結果得知甘草次酸能與胰島素接受器(insulin receptor)結合，進而利用抑制劑及西方墨點法分析甘草次酸其誘發解毒酵素之途徑，結果得知，甘草次酸能夠引發一連串的訊息傳遞，使PI3K/Akt/ mTOR路徑之激酶及ERK激酶，分別在20分鐘及60分鐘有磷酸化作用增加的現象，促使轉錄因子Nrf2轉位至細胞核內且結合於抗氧化反應單元的啟動子(Antioxidation respone element, ARE)序列，藉此活化轉錄使解毒酵素表現。 關鍵字：甘草次酸、Phase II解毒酵素、麩胱甘肽(GSH)、Nrf2 【Abstract】 Licorice root is a traditional herbal remedy that has been used for the treatment of various pathologic conditions, including harmonize with other drug, gastric ulcers, anti-hepatotoxic, anti-inflammatory and to relieve a cough. Its hydrolyzed metabolite glycyrrhetic acid reveal antihepatotoxic , anti-inflammatory and modulate immunity effects. The liver is the most important organ in the body involved in detoxification. Liver detoxification involves a number of enzymatic to metabolize the toxic substances which renders them less harmful and helps to facilitate their removal from the body. This study to investigate the effect of glycyrrhetic acid on the inducation of phase II enzyme in HepG2 cell line. Result indicate the glycyrrhetic acid may induce phase II detoxify enzyme, including Glutamate cysteine ligase (GCLc), Glutathione-S- transferase (GST), they get the morst expression by treat with glycyrrhetic acid for 12hr. Then GCLc is the important enzyme in Glutathion (GSH) synthesis. However, we obtained results showing that glycyrrhetic acid-induced increase in the GSH level and GST activity at 12 hr. In addition, flow cytometry assay that glycyrrhetic acid can binding to insulin receptor.