目的: 研究苦参碱诱导人肝癌BEL-7402细胞自吞噬和自吞噬性死亡作用。方法: 体外培养人肝癌BEL-7402细胞, 应用MTT法测定细胞增殖抑制率, 流式细胞术测定细胞凋亡, 自噬体标记物LC3免疫荧光定量检测自吞噬, 透射电镜分析细胞自吞噬死亡超微结构改变。结果: 苦参碱抑制细胞增殖呈质量浓度依赖性, 作用48h的IC50值为1.1mg/mL;0.6～1.6mg/mL苦参碱作用12h可诱导BEL-7402细胞凋亡逐渐增多, 1.2mg/mL作用12h时细胞凋亡率达(44.88±0.78)%; 1.2mg/mL苦参碱持续作用可以诱导自吞噬阳性细胞逐渐增多, 作用12h时自吞噬阳性细胞达(63.16±0.29)%; 超微结构显示细胞凋亡呈现自噬性细胞死亡特点。结论: 苦参碱体外能诱导BEL-7402细胞自吞噬和自噬性死亡。

Objective: To study the effects of matrine on inducing autophagy and autophagic death in BEL-7402 cells. Methods: BEL-7402 cells were cultured in RPMll640 alone or exposed to different concentration of matrine. Cell growth inhibition was assessed by MTT assay. Apoptosis was evaluated by flow cytometry. Autophagosome marker LC3 was examined by immunofluorescence microscopy. The ultrastructure change of ceils was analyzed by transmission electron microscope. Results: The cell proliferation was inhibited by matrine in a dose dependent manner. Its IC50value was 1.1 mg/mL at 48h. Treatment with 0.6-1.6 mg/mL matrine for 12h induced apoptosis of BEL-7402 cells and the apoptosis rate reached (44.88士0.78)% at concentration of 1.2 mg/mL. The autophagy positive cells were greatly increased after 1.2 mg/mL matrine treated in BEL-7402 cells and the number of cells reached(63.16±0.29)％. Morphological features of typical autophagic death were apparent in the cytoplasm at the uitrastructural level. Conclusion: Matrine could induce autophagy and autophagic death of BEL-7402 cells in vitro.

国家自然科学基金资助项目(30670688);国家环保公益性行业科研专项经费项目(2008467115)