目的 探究氧化苦参碱诱导结肠癌HCT116细胞死亡的作用机制。方法 将结肠癌HCT116细胞随机分为对照组和氧化苦参碱（2、4、8、12、16、20、30、40 mmol·L^-1）组，通过CCK-8法检测各浓度氧化苦参碱作用24、48 h对结肠癌细胞存活率的影响；光学显微镜下观察氧化苦参碱（15、20、25 mmol·L^-1）对HCT116细胞形态的影响；通过Hoechst染色法和Annexin V/PI双染流式细胞术法观察氧化苦参碱对HCT116细胞凋亡的影响；JC-1染色观察细胞线粒体膜电位的变化；Western blotting法检测各组细胞成熟体半胱氨酸蛋白酶-3（cleaved Caspase-3）、B淋巴细胞瘤-2（Bcl-2）、BCL2关联X蛋白（Bax）、细胞色素C （Cyt-C）、消化道皮肤素E （GSDME）、细胞周期蛋白1（cyclin-D1）、骨髓细胞瘤病毒癌基因（c-Myc）、聚腺苷二磷酸核糖聚合酶（PARP）、β-连环素（β-catenin）、生存素（survivin）的蛋白表达情况。结果 与对照组比较，氧化苦参碱可以明显降低HCT116细胞的存活率，且存在剂量及时间相关性，作用24、48h的半数抑制浓度（IC₅₀）分别为18.80、10.75 mmol·L^-1；氧化苦参碱组形状正常的HCT116细胞明显减少，均出现部分类圆球形的细胞，并且折光率明显降低；经Hoechst染色后氧化苦参碱组HCT116细胞的蓝色荧光明显加深，且相对集中，流式细胞术结果显示细胞凋亡率显著增加（P<0.05、0.01）；JC-1染色结果表明氧化苦参碱可以降低HCT116细胞线粒体膜电位；氧化苦参碱组Bcl-2、β-catenin、c-Myc、cyclin D1和survivin蛋白表达水平显著下降（P<0.05、0.01），Bax、cleaved Caspase-3、cleaved PARP、Cyt-C、GSDME蛋白表达水平及Bax/Bcl-2显著上升（P<0.05、0.01）。结论 氧化苦参碱可能通过下调Wnt/β-catenin信号通路，促进线粒体依赖的细胞内源性凋亡和细胞焦亡发挥抗结肠癌作用。

Objective To explore the mechanism of oxymatrine inducing colon cancer HCT116 cell death.Methods Colon cancer cells were randomly divided into control group, oxymatrine(2, 4, 8, 12, 16, 20, 30, and 40 mmol·L^-1) group.The CCK-8 method was used to detect the effect of different concentrations of oxymatrine on colon cancer cells survival after 24 and 48 h.The effects of oxymatrine(15, 20, 25 mmol·L^-1) on the morphological changes of HCT116 cells were observed by observation under optical microscope.Hoechst staining and Annexin V/PI double staining flow cytometry were used to observe the effect of oxymatrine on apoptosis of HCT116 cells.JC-1 staining was used to observe the changes of mitochondrial membrane potential.Western blotting method was used to detect the effect of oxymatrine on expressions of cleaved Caspase-3, B lymphocytoma-2(Bcl-2), BCL2-associated X protein(Bax), cytochrome C(Cyt-C), gastrointestinal dermatin E(GSDME), Cyclin 1(cyclin-D1), myeloma virus oncogene(c-Myc), poly adenosine diphosphate-ribose polymerase(PARP), β-catenin, survivin in colon cancer cells.Results Compared with control group, oxymatrine could significantly reduce the survival rate of HCT116 cells, and there was a dose and time correlation.The half inhibition concentration(IC50) of 24 and 48 h after treatment was 18.80 and 10.75 mmol·L^-1, respectively.In oxymatrine group, HCT116 cells with normal shape were significantly reduced, with some spherical like cells, and the refractive index was significantly reduced.After Hoechst staining, the blue fluorescence of HCT116 cells in oxymatrine group was significantly deepened and concentrated.Flow cytometry showed that the apoptosis rate was significantly increased(P<0.05, 0.01).JC-1 staining showed that oxymatrine could reduce the mitochondrial membrane potential of HCT116 cells.In oxymatrine group, the expression levels of Bcl-2, β-catenin, c-Myc, cyclin D1 and survivin protein decreased significantly(P<0.05, 0.01), while the expression levels of Bax, cleaved caspase-3, cleaved PARP, Cyt-C, GSDME protein and Bax/Bcl-2 protein increased significantly(P<0.05, 0.01).Conclusion Oxymatrine may play an anti-tumor effect by downregulating the Wnt/β-catenin signaling pathway and promoting mitochondria-dependent endogenous apopto sis and pyroptosis.

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江苏卫生健康职业学院院级重点课题（JKB202005）；江苏省大学生创新创业训练计划项目（202014255002Y）；江苏卫生健康职业学院院级重点课题（JKB2021005）；江苏卫生健康职业学院重大项（JKA202004）