目的：探讨新补骨脂异黄酮（NBIF）对肝细胞癌（HCC）Huh-7细胞焦亡的影响及其分子机制。方法：体外培养Huh-7细胞，用CCK-8法检测不同浓度的NBIF处理48 h时对细胞存活率的影响，光学显微镜下观察NBIF处理后Huh-7细胞的形态变化，乳酸脱氢酶（LDH）释放实验检测细胞的LDH 释放量，WB 实验检测细胞中GSDME、caspase-3 的蛋白水平变化。采用siRNA干扰Huh-7细胞中caspase-3、GSDME表达后，CCK-8法检测NBIF处理对细胞存活率的影响，WB实验检测GSDME蛋白表达水平，观察NBIF处理对细胞形态的影响，并检测细胞LDH释放量。结果：60 μmol/L以上的NBIF均能显著抑制Huh-7细胞的增殖（均P<0.01），光学显微镜下观察到NBIF处理后的细胞出现肿胀、吐泡现象，且LDH释放增加（P<0.01）；WB实验结果表明，NBIF能够激活caspase-3蛋白并切割GSDME蛋白，增加GSDME-N的表达（均P<0.01）。干扰caspase-3、GSDME表达后，NBIF对细胞的抑制作用减弱（均P<0.01），GSDME-N蛋白表达受到抑制（P<0.01），显微镜下细胞肿胀、吐泡现象几乎消失，LDH释放明显减少（P<0.05）。结论：NBIF能够通过caspase-3/GSDME途径诱导Huh-7细胞发生焦亡，从而抑制HCC细胞的增殖，为HCC的治疗提供一种新思路。

Objective: To explore the effect of neobavaisoflavone (NBIF) on pyroptosis of hepatocellular carcinoma (HCC) Huh-7 cells and its molecular mechanism. Methods: Huh-7 cells were cultured in vitro, and the effects of different concentrations of NBIF on cell survival rate was detected by CCK-8. Morphological changes of Huh-7 cells treated with NBIF were observed under optical microscope.Lactate dehydrogenase (LDH) release test was used to detect the LDH release of cells. The protein levels of gasdermin E (GSDME) and caspase-3 in the cells were detected by WB assay. After silencing caspase-3 and GSDME in Huh-7 cells, the effect of NBIF treatment on cell survival rate was detected by CCK-8. The expression level of GSDME protein was detected by WB assay. The effect of NBIF treatment on cell morphology was observed, and the release of LDH was detected. Results: NBIF above 60 μmol/L could significantly inhibit the proliferation of Huh-7 cells (all P<0.01). The swelling and bubbling phenomenon of NBIF treated cells were observed under optical microscope, and the release of LDH was increased (all P<0.01). WB results showed that NBIF could activate caspase-3 protein, cleave GSDME protein and increase the expression of GSDME-N (P<0.01). After silencing caspase-3 and GSDME genes, the inhibition of NBIF on cells was weakened (all P<0.01), and the expression of GSDME-N protein was inhibited (P<0.01). Under the microscope, the phenomenon of cell swelling and bubbling almost disappeared, and the release of LDH decreased significantly (P<0.05). Conclusion: NBIF can cause pyroptosis of Huh-7 cells through caspase3-GSDME pathway, thus inhibiting the proliferation of Huh-7 cells, which provides a new idea for the treatment of HCC.

吉林省教育厅科学技术研究项目（No. JJKH20220873KJ）；“杏林学者工程”青年科学家培养计划项目（No. QNKXJ2-2021ZR13）；吉林省青年科技人才托举工程项目（No. QT202111）