目的：中國醫學治療肝病已有千年以上的歷史，卻未能以現代科學證明，其優點隱而不現，本研究將依典籍論述的中醫辨證法，經適當與合理的轉譯方式，選取適當的方劑：左金丸，進行治療肝癌的研究，期使中醫臨床經驗與科學研究達到相輔相成的境界。
材料與方法：以左金丸及黃連、吳茱萸及其有效成份化合物，分析重組的肝癌母細胞(HepG2/AP-1 和HepG2/NF-κB)與非腫瘤肝細胞(Chang /AP-1)，在12-鄰-四葵酸-佛波醇-13-乙酸誘發肝癌細胞轉形的癌化過程中，觀察AP-1和NF-κB，表現量與其相關途徑表現。
結果：第一、由TPA引起的AP-1和/或NF-κB活化上升，伴隨肝癌母細胞轉形。當分別加入左金丸及黃連、吳茱萸及小蘗鹼與吳茱萸鹼活性化合物可抑制HepG2肝癌母細胞的AP-1和/或NF-κB的表現活化，阻斷癌化過程。第二、甲醇的吳茱萸萃取物，可有效抑制Chang /AP-1細胞株的細胞轉形，比較三種主要活性化物吳茱萸鹼、吳茱萸次鹼和檸檬苦素，發現吳茱萸鹼可有效抑制AP-1活性，分析其訊息傳遞路徑為MAPK/ERKs。
結論：不論是左金丸或是吳茱萸、黃連，以及活性成份小蘗鹼和吳茱萸鹼都可抑制HepG2肝癌母細胞的癌化過程；其中左金丸與黃連小蘗鹼是透過阻斷AP-1和NF-κB路徑。而吳茱萸與吳茱萸鹼可抑制HepG2肝細胞的細胞轉形的過程，是降低AP-1活性。在Chang liver細胞中發現：其機轉是降低AP-1活性經MAPK的ERKs的磷酸化減少。另外比較肝癌母細胞(HepG2)與非腫瘤細胞(Chang liver cell)都可被吳茱萸與吳茱萸鹼所抑制，可見吳茱萸鹼可針對被誘發的AP-1活性的肝細胞，有選擇性的抑制癌化過程中的tumor promotion。
PURPOSE: Zuo-Jin-Wan (ZJW) has been used to treat digestive diseases in Asia. This study was to determine whether ZJW and its components blocked activator protein 1(AP-1) and/ or nuclear factor-κB (NF-κB) activities as well as tumor promotion in Chang liver cells and hepatoblastoma HepG2 cells.
MATHODS AND MATERIALS: The Chang/AP‐1 and HepG2/ AP-1 and HepG2/ NF-κB cells were treated with the AP‐1 and NF-κB stimulator 12‐O‐tetradecanoylphorbol‐13‐ acetate. And ZJW, Coptis chinensis, Evodia rutaecarpa and its components, berberine, evodiamine, rutaecarpine and limonin were analyzed in anchorage-independent growth of Chang liver or HepG2 cells.
RESULTS: ZJW and its components, Coptis chinensis and Evodia rutaecarpa, inhibited AP-1 and/ or NF-κB activities, and suppressed anchorage-independent growth of Chang liver and/ or HepG2 cells. The major alkaloidal ingredients, berberine and evodiamine, inhibited AP-1 activities and/or NF-κB activation, and further suppressed hepatocellular transformation. Moreover, evodiamine significantly diminished the TPA‐induced phosphorylation of extracellular signal‐regulated kinases. These results suggested that evodiamine treatment suppressed the TPA‐induced AP‐1 activity via the ERKs pathway.
CONCLUSION: ZJW, Coptis chinensis, Evodia rutaecarpa, berberine and evodiamine inhibited the AP‐1 and/ or NF-κB activity and cellular transformation in human hepatocytes, suggesting that ZJW and its active components were a potential agents for antitumor therapy.
