[摘要] 目的：探究积雪草酸（asiatic acid，AA）对人肝癌细胞恶性生物学行为的抑制作用及其机制。方法：选择人肝癌Huh7细胞作为研究对象，体外培养过程中分别给予0、5、10、25、50、100 μmol/L的AA处理，采用CCK-8 法和EdU实验测定其对细胞增殖的影响，流式细胞术检测凋亡和细胞周期的变化，WB检测凋亡相关蛋白p-AKT、p-ERK1/2、p-p38、cleaved-caspase3、cleavedcaspase9、BAX、Bcl-2、AKT、ERK、p38、pro-caspase3 和pro-caspase9 表达的变化。结果：AA能够以剂量和时间依赖的方式抑制Huh7 细胞的增殖（均P<0.05）；10 μmol/LAA孵育Huh7 细胞24 h 后细胞增殖被显著抑制（P<0.05）、凋亡率显著升高（P<0.05）、细胞周期阻滞于G1 期（P<0.05）。AA可诱导p-p38 的表达，但以剂量依赖的方式抑制p-AKT和p-ERK的表达（均P<0.05）；随着AA浓度的增加，cleaved-caspase3、cleaved-caspase9、BAX的水平增加，而Bcl-2 的水平降低（均P<0.05）。结论：AA抑制人肝癌细胞的增殖并促使其凋亡，该作用与MAPK和PI3K/AKT通路有关。

[Abstract] Objective: To investigate the inhibitory effect of asiatic acid (AA) on malignant biological behaviors of human liver cancer cells and to explore the mechanism. Methods: Human liver cancer cell line (Huh7) was used as research subject, and treated with different concentrations of AA (0, 5, 10, 25, 50, 100 μmol/L) in vitro. The effect of AA on cell proliferation was determined by CCK-8 and EdU assay; the apoptosis and cell cycle distribution were detected by flow cytometry, while the expressions of apoptosis-related proteins (AKT, P-ERK 1/2 , p38, cleaved-caspase3, cleaved-caspase9, BAX, Bcl-2, AKT, ERK, p38, pro-caspase 3 and pro-caspase 9)were examined by WB. Results: AA could inhibit the proliferation of Huh7 cells in a dose- and time-dependent manner (all P<0.05).After being incubated with 10 μmol/L AA for 24 h, the proliferation of Huh7 cells was significantly inhibited (P<0.05), the apoptosis rate was significantly increased (P<0.05), and cell cycle was arrested in G1 phase (P<0.05). AA induced p-p38 expression, but inhibited the expression of p-AKT and p-ERK in a dose-dependent manner (all P<0.05). In addition, as the concentration of AA increased, the levels of cleaved-caspase 3, cleaved-caspase 9 and BAX increased, while the level of Bcl-2 decreased (all P<0.05). Conclusion: AA inhibits the proliferation of human liver cancer cells and promotes its apoptosis, which is associated with the MAPK and PI3K/AKT pathways.

国家自然科学基金资助（No. 81430101）