An Ning, Wang Xiaoru, Qin Jia, Cheng Meng, Bai Mei, Cheng Jingcai, Xu Qiang, Wu Xuefeng
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing Drum Tower Hospital, School of Life Sciences, Nanjing University, Nanjing, China.
Drug R&D Institute, JC (Wuxi) Company, Inc., Wuxi, China.
Front Pharmacol. 2025 Mar 25;16:1539934. doi: 10.3389/fphar.2025.1539934. eCollection 2025.
Drug-induced liver injury (DILI) represents a distinct form of hepatic damage resulting from exposure to pharmacological agents. The pathological mechanisms underlying DILI are multifaceted and remain incompletely elucidated. However, emerging evidence suggests that cell pyroptosis, a form of programmed cell death associated with inflammation, may serve as a common mechanistic pathway in DILI pathogenesis. To investigate the role of pyroptosis in DILI, we established a murine model of DILI using triptolide and evaluated the therapeutic potential of phenethyl isothiocyanate (PEITC), a naturally occurring compound, in mitigating liver injury through the modulation of hepatocyte pyroptosis. Mice were administered PEITC at doses ranging from 5 to 20 mg/kg. Cytokine expression was measured using quantitative polymerase chain reaction or biochemical indicator analyzer. Cell signalings were assayed by western blot and immunohistochemistry. The AML12 hepacytes were cultured to investigate the effects. PEITC treatment markedly attenuated hepatic tissue damage, restored normal liver architecture, and significantly reduced serum levels of transaminases (AST and ALT), while normalizing hepatic metabolic function. These protective effects were mechanistically linked to the suppression of hepatocyte pyroptosis, as PEITC effectively reversed the upregulation of the NLRP3 inflammasome, Caspase-1 cleavage, and Gasdermin D (GSDMD) in triptolide-exposed livers. studies using cultured hepatocytes further demonstrated that PEITC inhibited the expression and activation of NLRP3, Caspase-1, GSDMD, and other key proteins involved in the pyroptosis pathway. Ultrastructural analysis via electron microscopy corroborated these findings, revealing that PEITC prevented pyroptosis-induced membrane pore formation in hepatocytes. PEITC exerts hepatoprotective effects against DILI by targeting the pyroptosis pathway, thereby highlighting its potential as a novel therapeutic strategy for liver injuries. Our results further implicate cell pyroptosis as a novel target for the attenuation of DILI.
药物性肝损伤（DILI）是由接触药理制剂引起的一种独特的肝损伤形式。DILI的病理机制是多方面的，仍未完全阐明。然而，新出现的证据表明，细胞焦亡作为一种与炎症相关的程序性细胞死亡形式，可能是DILI发病机制中的一个共同机制途径。为了研究焦亡在DILI中的作用，我们使用雷公藤内酯醇建立了DILI小鼠模型，并评估了天然化合物异硫氰酸苯乙酯（PEITC）通过调节肝细胞焦亡减轻肝损伤的治疗潜力。给小鼠施用5至20mg/kg剂量的PEITC。使用定量聚合酶链反应或生化指标分析仪测量细胞因子表达。通过蛋白质印迹和免疫组织化学分析细胞信号传导。培养AML12肝细胞以研究其作用。PEITC治疗显著减轻了肝组织损伤，恢复了正常的肝脏结构，并显著降低了转氨酶（AST和ALT）的血清水平，同时使肝脏代谢功能正常化。这些保护作用在机制上与抑制肝细胞焦亡有关，因为PEITC有效地逆转了雷公藤内酯醇暴露肝脏中NLRP3炎性小体、半胱天冬酶-1裂解和Gasdermin D（GSDMD）的上调。使用培养的肝细胞进行的研究进一步表明，PEITC抑制了焦亡途径中涉及的NLRP3、半胱天冬酶-1、GSDMD和其他关键蛋白的表达和激活。通过电子显微镜进行的超微结构分析证实了这些发现，表明PEITC可防止肝细胞中焦亡诱导的膜孔形成。PEITC通过靶向焦亡途径对DILI发挥肝保护作用，从而突出了其作为肝损伤新治疗策略的潜力。我们结果进一步表明细胞焦亡是减轻DILI的新靶点。
