目的 探讨桃红四物汤改善硫代乙酰胺（thioacetamide，TAA）诱导的肝细胞新型死亡模式——双硫死亡，从而干预肝纤维化进展的作用机制。方法 利用高效液相色谱仪对桃红四物汤进行成分分析。建立TAA诱导的小鼠肝纤维化模型和AML12肝细胞模型，给予桃红四物汤干预后，采用转录组测序、生物信息学、qRT-PCR和免疫荧光法探究桃红四物汤抑制肝细胞双硫死亡的作用机制。结果 TAA可诱导双硫死亡标志基因如溶质载体家族7成员11（solute carrier family 7 member 11，Slc7a11）、溶质载体家族3成员2（solute carrier family 3 member 2，Slc3a2）、细丝蛋白A（filamin A，Flna）、肌球蛋白重链9（myosin heavy chain 9，Myh9）、肌球蛋白重链10（myosin heavy chain 10，Myh10）、NCK关联蛋白1（NCK associated protein 1，Nckap1）及纤维化相关基因如肌动蛋白α2（actin alpha 2，Acta2）、I型胶原蛋白α1链（collagen type I alpha 1 chain，Col1a1）、金属蛋白酶组织抑制剂1（tissue inhibitor of metalloproteinase 1，Timp1）表达显著上调（P＜0.05、0.01、0.001），伴随F-actin收缩及烟酰胺腺嘌呤二核苷磷酸（nicotinamide adenine dinucleotide phosphate，NADP⁺）/NADPH值升高（P＜0.01）。给予桃红四物汤干预后，细胞骨架基因Flna、Myh9、Myh10、Nckap1及肝纤维化相关基因表达显著下调（P＜0.05、0.01、0.001），NADP⁺/NADPH值显著降低（P＜0.05），细胞骨架的形态恢复。结论 桃红四物汤可通过有效缓解TAA诱导的肝细胞双硫死亡延缓肝纤维化进展，为相关疾病及并发症的临床治疗提供理论依据。

Objective To investigate the mechanism by which Taohong Siwu Decoction (桃红四物汤) ameliorates thioacetamide (TAA)-induced liver fibrosis via intervening in hepatocyte disulfidptosis, a novel model of hepatocyte death. Methods High performance liquid chromatography was used to analyze the components of Taohong Siwu Decoction. TAA-induced mouse liver fibrosis model and AML12 liver cell model were established, and after intervention with Taohong Siwu Decoction, the mechanism of Taohong Siwu Decoction in inhibiting hepatocyte disulfidptosis was investigated using transcriptome sequencing, bioinformatics, qRT-PCR and immunofluorescence methods. Results TAA could induce the up-regulation of disulfidptosis marker genes such as solute carrier family 7 member 11 (Slc7a11), solute carrier family 3 member 2 (Slc3a2), filamin A (Flna), myosin heavy chain 9 (Myh9), myosin heavy chain 10 (Myh10), NCK associated protein 1 (Nckap1) and fibrosis related genes such as actin alpha 2 (Acta2), collagen type I alpha 1 chain (Col1a1), tissue inhibitor of metalloproteinases 1 (Timp1) expressions (P < 0.05, 0.01, 0.001), accompanied by F-actin contraction and nicotinamide adenine dinucleotide phosphate (NADP⁺)/NADPH value increased (P < 0.01). After intervention with Taohong Siwu Decoction, the expressions of cytoskeletal genes Flna, Myh9, Myh10, Nckap1 and liver fibrosis related genes were significantly down-regulated (P < 0.05, 0.01, 0.001), and NADP⁺/NADPH value was significantly reduced (P < 0.05), the morphology of cytoskeleton was restored. Conclusion Taohong Siwu Decoction could effectively alleviate TAA-induced hepatocyte disulfidptosis and delay the progression of liver fibrosis, providing a theoretical basis for the clinical treatment of related diseases and complications.

R285.5

国家自然科学基金青年项目（82404984）；中央高校基本科研业务费专项资金（2024-JYB-JBZD-055）