目的 探究香芹酚对肝星状细胞（hepatic stellate cell，HSC）铁死亡的影响及其抗肝纤维化的作用机制。方法 体内实验以四氯化碳诱导的小鼠肝纤维化模型为研究对象，设置对照组、模型组和香芹酚低、中、高剂量（25、50、100 mg/kg）组。采用苏木素-伊红（hematoxylin-eosin，HE）和Masson染色观察各组小鼠肝组织病理变化；采用试剂盒检测小鼠血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天冬氨酸氨基转移酶（aspartate aminotransferase，AST）活性和羟脯氨酸（hydroxyproline，HYP）水平；采用Western blotting检测小鼠肝组织谷胱甘肽过氧化物酶4（glutathione peroxidase 4，GPX4）、α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、I型胶原α1（collagen type 1α1，Col1α1）蛋白表达。体外实验以HSC-T6细胞为研究对象，用不同浓度（100、200、300、400、500、600 μmol/L）的香芹酚或铁死亡抑制剂Ferrostatin-1（Fer-1）或缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）稳定剂DMOG（1 mmol/L）进行干预，CCK-8检测细胞活力；试剂盒检测HSC-T6细胞中Fe²⁺、谷胱甘肽（glutathione，GSH）、丙二醛（malondialdehyde，MDA）和活性氧（reactive oxygen species，ROS）含量；Western blotting检测HSC-T6细胞中GPX4、α-SMA、Col1α1、HIF-1α、溶质载体家族7成员11（solute carrier family 7 member 11，SLC7A11）蛋白表达。结果 与模型组比较，香芹酚组小鼠血清中ALT、AST活性和HYP水平显著降低（P＜0.05、0.01），肝脏肿胀、炎性细胞浸润和胶原沉积减少，肝组织α-SMA、COL1α1和GPX4蛋白表达水平显著降低（P＜0.05、0.01）。香芹酚可降低HSC-T6细胞中GSH水平（P＜0.05、0.01），并提高Fe²⁺、MDA、ROS的含量（P＜0.05、0.01），其对HSC-T6细胞活力的抑制作用可被Fer-1逆转（P＜0.05、0.01）；香芹酚对HSC-T6细胞中HIF-1α总蛋白水平无显著影响，但可降低细胞核中的HIF-1α水平以及细胞SLC7A11总蛋白水平（P＜0.01）；HIF-1α稳定剂DMOG可阻断香芹酚对HSC-T6细胞铁死亡的诱导作用（P＜0.05、0.01）。结论 香芹酚能够通过HIF-1α/SLC7A11轴诱导HSC铁死亡，进而发挥抗肝纤维化的作用。

Objective To explore the effect of carvacrol on ferroptosis in hepatic stellate cell (HSC) and its mechanism against liver fibrosis. Methods In vivo experiment was conducted on a mouse liver fibrosis model induced by carbon tetrachloride. The control group, model group, and carvacrol low-, medium-, high-dose (25, 50, 100 mg/kg) groups were set up, hematoxylin-eosin (HE) and Masson staining were used to observe the pathological changes of liver tissue in each group of mice; Kits were used to detect the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hydroxyproline (HYP) level in serum of mice; Western blotting was used to detect the protein expressions of glutathione peroxidase 4 (GPX4), α-smooth muscle actin (α-SMA) and collagen type 1α1 (Col1α1) in liver tissue of mice. In vitro experiments were conducted on HSC-T6 cells, and different concentrations (100, 200, 300, 400, 500, 600 μmol/L) of carvacrol or ferroptosis inhibitor ferrostatin-1 (Fer-1) or hypoxia inducible factor-1α (HIF-1α) stabilizer DMOG (1 mmol/L) were used for intervention. Cell viability was detected by CCK-8; The reagent kits were used to detect the levels of Fe²⁺, glutathione (GSH), malondialdehyde (MDA) and reactive oxygen species (ROS) in HSC-T6 cells; Western blotting was used to detect the protein expressions of GPX4, α-SMA, Col1α1, HIF-1α and solute carrier family 7 member 11 (SLC7A11) in HSC-T6 cells. Results Compared with model group, the activities of ALT, AST and level of HYP in serum of mice in carvacrol group were significantly reduced (P < 0.05, 0.01), while liver swelling, inflammatory cell infiltration and collagen deposition were reduced; The protein expression levels of α-SMA, COL1α1 and GPX4 in liver tissue were significantly reduced (P < 0.05, 0.01). Carvacrol could reduce GSH level in HSC-T6 cells (P < 0.05, 0.01) and increase the contents of Fe²⁺, MDA and ROS (P < 0.05, 0.01); Its inhibitory effect on HSC-T6 cell viability could be reversed by Fer-1 (P < 0.05, 0.01); Carvacrol had no significant effect on the total protein level of HIF-1α in HSC-T6 cells, but could reduce the HIF-1α level in nucleus and the total protein level of SLC7A11 in HSC-T6 cells (P < 0.01); The HIF-1α stabilizer DMOG could block the induction effect of carvacrol on ferroptosis in HSC-T6 cells (P < 0.05, 0.01). Conclusion Carvacrol can induce ferroptosis in HSC through HIF-1α/SLC7A11 axis, thereby exerting an anti-liver fibrosis effect.

R285.5

国家自然科学基金青年科学基金项目（82004010）;安徽省教育厅重点项目（2022AH051163）;安徽省重点研发计划项目（2022e07020061）