[摘 要] 目的：探究蟾毒灵（Buf）对卵巢癌细胞A2780和SKOV3的毒性作用及其机制，评估其与顺铂（DDP）联合应用的协同毒性。方法：常规培养A2780和SKOV3细胞。用CCK-8法、克隆形成实验检测Buf对A2780和SKOV3细胞增殖的影响及半数抑制浓度（IC50）。用不同浓度的Buf处理细胞，显微镜下观察细胞形态变化，比色法检测乳酸脱氢酶（LDH）释放水平，流式细胞术分析检测Annexin Ⅴ+PI+细胞比例，qPCR和WB法检测细胞焦亡相关分子mRNA及蛋白表达水平，通过小干扰RNA敲减A2780和SKOV3细胞中的GSDMD并进行后续功能实验进行验证。通过SynergyFinder分析Buf与DDP对于杀伤A2780和SKOV3细胞的协同效应，并通过流式细胞术、qPCR、WB进行机制探索，进一步利用流式细胞术检测Buf与DDP联用对DDP耐药A2780和SKOV3细胞的影响。结果：与对照组相比，Buf显著抑制卵巢癌细胞增殖及迁移能力（均P < 0.05），诱导LDH释放和细胞焦亡（均P < 0.05），上调核苷酸结合寡聚化结构样受体热蛋白结构域相关蛋白3（NLRP3）、半胱氨酸天冬氨酸蛋白水解酶1（CASP1）、gasdermin家族成员D（GSDMD）、白细胞介素-1β（IL-1β）及IL-18的mRNA及蛋白的表达（均P < 0.05），Buf与DDP联用具有协同抑制A2780和SKOV3细胞增殖的作用，并可进一步增强DDP耐药的A2780和SKOV3细胞对DDP的敏感性（P < 0.01或P < 0.001）。结论：Buf可通过激活NLRP3/CASP1/GSDMD信号通路诱导A2780和SKOV3细胞焦亡，并增强A2780和SKOV3细胞及DDP耐药A2780和SKOV3细胞对DDP的敏感性。

ObjectiveTo investigate the clinical effect of Jiedu Liangxue Jianpi prescription in the treatment of patients with hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF) in early and middle stages. MethodsA total of 75 patients who were diagnosed with HBV-ACLF in early and middle stages in Beijing Ditan Hospital from October 2017 to January 2019 were enrolled and randomly divided into integrated traditional Chinese and Western medicine group with 36 patients and control group with 39 patients. The patients in the control group were given Western medicine treatment, and those in the integrated traditional Chinese and Western medicine group were treated with Jiedu Liangxue Jianpi prescription in addition to Western medicine treatment. The course of treatment was 8 weeks for both groups. Cumulative response rate was observed for both groups, and the two groups were compared in terms of liver function, coagulation function, and routine blood test results before and after treatment. A univariate or multivariate analysis of variance with repeated measures was used for comparison between groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsAt week 4 of treatment, the integrated traditional Chinese and Western medicine group had a significantly higher cumulative response rate than the control group (77.7% vs 53.8%, χ2=4.734, P＜0.05). Compared with the control group, the integrated traditional Chinese and Western medicine group had a significant increase in albumin at weeks 1 and 2 of treatment (all P＜0.05), a significant increase in prothrombin activity at week 1 of treatment (P＜0.05), and significant reductions in neutrophil count and neutrophil-lymphocyte ratio at weeks 3 and 8 of treatment (all P＜0.05). ConclusionJiedu Liangxue Jianpi prescription can improve the treatment response, liver function, and coagulation function and reduce neutrophil-lymphocyte ratio in patients with HBV-ACLF in early and middle stages.