目的 探讨吴茱萸碱（evodiamine，Evo）对结肠癌HCT-116细胞自噬、增殖的影响及其可能的分子机制。方法 CCK-8法检测Evo对HCT-116细胞增殖的影响；Evo（3、6 μmol/L）处理48 h后，MDC法检测细胞内自噬小泡的数量，DHE法检测细胞内活性氧（ROS）的量，Western blotting法检测细胞自噬相关蛋白和腺苷酸活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）通路相关蛋白的表达；Evo（6 μmol/L）分别与自噬抑制剂3-甲基腺嘌呤（3-methyladenine，3-MA）和凋亡抑制剂Z-DEVD-FMK共同作用48 h后，Western blotting法检测细胞内自噬及凋亡相关蛋白的表达。结果 与对照组比较，Evo对HCT-116细胞呈现浓度依赖性的增殖抑制作用；Evo（3、6 μmol/L）使HCT-116细胞内的ROS量升高、自噬小泡数量增加、自噬相关蛋白LC3表达增加、p62表达降低、p-AMPK及mTOR表达增加；Evo与自噬抑制剂联合给药后，细胞内LC3蛋白表达减少，而有活性的Caspase-3表达增加，Evo与凋亡抑制剂联合给药后，细胞内LC3表达增加，而有活性的Caspase-3表达被抑制。结论 Evo能够通过AMPK/mTOR通路激活自噬、抑制HCT-116细胞增殖，且细胞自噬与凋亡呈现互补作用。

Objective To investigate the effect of evodiamine (Evo) on the autophagy and proliferation of colon cancer HCT-116 cells and the underlying mechanism. Methods The effect of Evo on proliferation of HCT-116 cells was detected by CCK-8 method. After being processed with Evo (3 and 6 μmol/L) for 48 h, the number of autophagic vesicles were detected by MDC method. The amount of ROS in HCT-116 cells was measured by DHE assay, and the protein related with autophagy and AMPK/mTOR pathway was detected by Western blotting. After the treatment of Evo (6 μmol/L) combined with autophagy inhibitor 3-MA (3-methyladenine) or apoptosis inhibitor Z-DEVD-FMK respectively for 48 h, Western blotting was used to detect the expression of autophagy and apoptosis-related protein in HCT-116 cells. Results Compared with the control group, Evo inhibited the proliferation of HCT-116 cells in a dose-dependent manner; After treated with Evo (3 and 6 μmol/L) for 48 h, the amount of intracellular ROS and autophagic vesicles were increased, the protein expression levels of LC3, p-AMPK, and mTOR were increased while the expression of p62 was increased. After being treated with Evo and autophagy inhibitor, the protein expression of LC3 was decreased while activated Caspase-3 was increased; Combination of Evo and apoptosis inhibitor increased the expression of LC3 and inhibited the expression of activated Caspase-3. Conclusion Evo can activate autophagy of HCT-116 cells through AMPK/mTOR pathway and inhibit the proliferation, and the effect of autophagy and apoptosis on cells are complementary.

国家自然科学基金资助项目（31271368）；重庆市渝中区科技计划项目（20140123）