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目的 观察顺铂对类毛细胞House耳研究所-耳蜗1型细胞（House Ear Institute-Organ of Corti 1,HEI-OC1）的损伤效应及毒性机制。方法 培养HEI-OC1细胞系，以顺铂构建损伤模型，以Caspase-1抑制剂VX-765联合干预，进行细胞活性检测，应用流式细胞术、乳酸脱氢酶释放检测、吖啶橙/溴化乙锭双荧光染色判断细胞膜通透性变化，观察细胞形态变化，分别以酶联免疫吸附试验试剂盒、蛋白质印迹法和免疫荧光法检测焦亡相关靶蛋白的定量及定位表达，定量逆转录聚合酶链反应法检测细胞内相应的mRNA的表达量。结果 与对照组比较，顺铂刺激组的HEI-OC1细胞存活率降低，细胞膜通透性增加，部分细胞呈现“气球样变”，细胞膨大变形，细胞膜破裂，焦亡相关靶蛋白及基因mRNA表达量升高（P<0.05），给予细胞焦亡抑制剂Caspase-1抑制剂（Belnacasan,VX-765）刺激后，细胞存活率增高，焦亡相关靶蛋白及基因mRNA表达量降低（P<0.05）。结论 顺铂以浓度依赖的方式诱导HEI-OC1细胞死亡，Caspase-1介导的细胞焦亡在顺铂诱导的耳毒性过程中发挥重要作用，这一作用可能是通过调节NLRP3/Caspase-1/GSDMD路径活化实现的。
Abstract：Objective To investigate damaging toxic effects of cisplatin on House Ear Institute-Organ of Corti 1(HEIOC1) cells and potential mechanisms. Methods In this study, a model of HEI-OC1 cell injury by cisplatin was established. The Caspase-1 inhibitor VX-765 was used as intervention. The CCK-8 method was used to identify cell viability. Flow cytometry, LDH release assay, and AO/EB fluorescence staining were used to examine cellular membrane integrity. ELISA, WesternBlot, immunofiuorescence and qRT-PCR were used to demonstrate expression of pyroptosis-related proteins and its mRNAs. Results Compared with the control, cisplatin treated cells demosntrated decreased survival, with some cells showing increased membrane permeability, swelling and deformation or cell membrane rupture. Expression of pyroptosis-related proteins and mRNAs was increased(P<0.05). Compared with cisplatin treatment only, cells treated with VX-765+cisplatin showed increased survival with decreased expression of pyroptosis-related proteins and mRNAs(P<0.05). Conclusions Cisplatin induced death of HEI-OC1 cells in a concentration-dependent manner. Caspase-1-mediated cellular pyroptosis plays an important role in cisplatin-induced ototoxicity, potentially by regulating the NLRP3/Caspase-1/GSDMD pathway.
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基本信息:
中图分类号:R969
引用信息:
[1]符昌学,张亚男,田美慧,等.基于NLRP3/Caspase-1/GSDMD通路探讨顺铂耳毒性的焦亡机制[J].中华耳科学杂志,2025,23(09):1045-1052.
基金信息:
吉林省卫生健康委员会项目（2022JC034）; 国家级大学生创新创业训练计划项目（202310199040）
2025-12-02
2025-12-02