中藥方劑小柴胡湯之成分誘發人類肝癌細胞計劃性死亡之機制探討

呂鋒洲 
國立台灣大
有關中藥方劑--小柴胡湯之臨床用途包括：保肝、抗發炎、抗過敏、改善腎障害、抗高血脂、抗放射線、改善氣喘以及免疫增強作用等等，回顧文獻指出，小柴胡湯可以藉由抑制自由基的產生、清除自由基等抗氧化能力來減緩組織細胞障害，調節基因表現進而阻止肝組織纖維化、肝硬化甚至肝癌的產生。因中藥方劑多為各單味藥材加以炮製煎煮而成，其藥效又因來源及藥性間相輔相佐而有協同加成的影響，考慮實驗設計上的單純化，本計畫擬以小柴胡湯之單味藥材黃芩的主要成分，如wogonin、baicalein與baicalin等化合物來處理各種人類的肝癌細胞株，模擬臨床上各療程面臨之癌細胞的狀況及種類，並探討這些成分對於細胞之效應，觀察其是否可抑制癌細胞的生長或藉由引起計畫性細胞凋亡之途徑來殺死癌細胞。本計畫首先以trypan blue exclusion assay與MTT assay評估黃芩的主要成分對肝癌細胞的細胞毒性與抑制肝癌細胞生長的濃度，根據預試驗的結果指出：wogonin、baicalein與baicalin等化合物的確可以有效抑制癌細胞生長甚至引起癌細胞之死亡。本計劃欲使用流式細胞儀技術針對各成分處理細胞後，在細胞層次的效應；流式細胞儀可以針對單一細胞進行偵測，若配合適當染劑，更可同時執行多種功能之評估，集準確與快速之優點，可以更有效率的整合藥物處理細胞後的影響，如：DNA的合成速率、細胞週期、細胞計劃性凋亡、粒線體功能、氧自由基之生成、細胞抗氧化能力等方面之評估。

關鍵字：黃芩、細胞計劃性凋亡、流式細胞儀

The mechanism of apoptosis induced by compounds isolated from the herbal medicine, sho-saiko-to.

Lu Fung-Jou
Department of Biochemistry, College of Medicine, National Taiwan University.
Sho-saiko-to is used as a herbal medicine in Chinese and Japanese community. This oral medicine consists of 7 herb components (ie, bupleurum root, pinellia tuber, scutellaria root, jujube fruit, ginger rhizome, ginseng root, and glycyrrhiza root) and has been used in treatment for pyretic disease as well as for gradually improving subjective symptoms and abnormal liver functions of patients with chronic viral liver diseases. For the reason of simplification and the preliminary data tested in our group, we would like to evaluate the mechanism of apoptosis induced by the active compound in scutellaria root, wogonin, by the technique of flowcytometry. The items proposed include the distribution of cell cycle, the function of mitochondria, the generation of reactive oxygen species and the cellular redox potential assessment. We would like to monitor the cellular alteration after the intervention of wogonin and most importantly to introduce more scientific experiments and evidences in this
關鍵字：Scutellaria root, apoptosis, flowcytometry