海南医科大学学报

目的：探讨6-姜酚(6-gingerol,6-G)调控酰基辅酶A合成酶4(ACSL4)/溶质载体7家族成员11 ,SLC7A11(xCT)/谷胱甘肽过氧化物酶4(GPX4)抑制ApoE~((-/-))动脉粥样硬化(atherosclerosis,AS)小鼠铁死亡的作用及机制。方法：6只普通饲料喂养C57BL/6J雄性小鼠为对照(Control,Con)组，按随机数字表法将18只SPF级别的ApoE~((-/-))小鼠分成AS模型(atherosclerosis Model,Mod)组，6-G治疗(6-Gingerol,6-G)组，铁死亡抑制剂(Ferrostatin-1,Fer-1)组，每组6只。Con组喂养普通饲料，除Con组，其余组别均喂养高脂饲料，持续喂养10周，诱导构建AS动物模型。造模成功后，6-G组给予20 mg/kg的6-G灌胃处理，Fer-1组给予1 mg/kg的Fer-1腹腔注射，除Con组仍饲喂普通饲料外，其余组仍继续饲喂高脂饲料，继续喂养6周。全自动生化分析仪检测血脂水平，油红O染色法检测小鼠动脉组织粥样硬化水平，透射电子显微镜观察各组主动脉线粒体超微形态结构改变，试剂盒检测Fe²⁺水平，ELISA法测定血清还原型谷胱甘肽(GSH)和丙二醛(MDA)的水平。Western blot评估小鼠主动脉ACSL4、xCT和GPX4蛋白表达情况。结果：与Con相比，Mod小鼠主动脉见明显脂质斑块附着，血清TC、TG、LDL-C、Fe²⁺、MDA水平升高，HDL-C、GSH水平降低(P<0.01)，主动脉组织ACSL4蛋白表达明显上升，GPX4、xCT表达明显下降(P<0.01)，线粒体结构形态损伤严重。与Mod组相比，6-G组以及Fer-1组小鼠治疗后主动脉脂质斑块沉积减少，血清TC、TG、LDL-C、Fe²⁺、MDA水平降低，HDL-C、GSH水平升高，差异具有统计学意义(P<0.05)，主动脉组织ACSL4蛋白表达降低，GPX4、xCT的蛋白表达量明显升高，差异具有统计学意义(P<0.05)，线粒体结构趋近正常。结论：6-姜酚可以减轻动脉粥样硬化水平，其作用机制可能是通过调节ACSL4/xCT/GPX4途径抑制铁死亡和脂质过氧化发挥对主动脉的保护作用。

Objective: To investigate the influence and underlying mechanisms of 6-gingerol(6-G) in suppressing ferroptosis in ApoE~((-/-)) atherosclerotic mice by modulating the ACSL4(Acyl-CoA synthetase 4)/xCT(solute carrier 7 family member 11, SLC7A11)/GPX4(glutathione peroxidase 4) signaling pathway. Methods: A total of 6 male C57 BL/6J mice were placed on a normal diet to serve as the control group(Con). A total of 18 SPF grade ApoE~((-/-)) mice were randomly assigned to the AS model group(Atherosclerosis Model group,Mod), the 6-G treatment group(6-Gingerol group,6-G), and the Ferrostatin-1 group(Ferrostatin-1 group,Fer-1), with 6 mice allocated to group. The Con group continued with a normal diet, while all other groups were subjected to a high-fat diet for a duration of 10 weeks to establish the AS animal model. Upon successful modeling, the 6-G group received 20 mg/kg of 6-G via gavage, while the Fer-1 group was administered 1 mg/kg~/of Fer-1 through intraperitoneal injection. Apart from the Con group, which consumed a normal diet, the other groups remained on the high-fat diet for an additional 6 weeks. Blood lipid profiles were assessed using an automatic biochemical analyzer, while atherosclerosis levels in the mice were evaluated through oil red O staining. The ultra-structural alterations of aortic mitochondria across all groups were examined using a transmission electron microscope. Fe²⁺ levels was detected using a reagent kit, while serum levels of GSH and MDA were quantified via ELISA.The protein levels of ACSL4, xCT, and GPX4 in the aortic tissues of the mice from each group were determined through Western blot analysis. Results: Compared to the Con group, the aortic tissues of Mod mice exhibited significant lipid plaque accumulation, along with elevated serum levels of TC,TG, LDL-C, Fe²⁺, and MDA was increased and HDL-C and GSH levels was decreased(P<0.01). The expression of ACSL4 protein in the aortic tissue was notably elevated, whereas the expression of GPX4 and xCT proteins was significantly reduced(P<0.01). Additionally, mitochondrial structure and morphology were severely compromised. Compare to the Mod group, both the 6-G group and the Fer-1 group exhibited a reduction in aortic lipid plaque accumulation following treatment. There was a notable decrease in the levels of serum TC, TG, LDL-C, Fe²⁺, and MDA, while levels of HDL-C and GSH were found to be elevated, with significant differences observed(P<0.05). Additionally, there was a down-regulation of ACSL4 protein expression, while GPX4 and xCT protein expressions were up-regulated, also showing significant differences(P<0.05), with mitochondrial structures nearing normalcy. Conclusion: 6-gingerol can mitigate the level of atherosclerosis, potentially through the modulation of the ACSL4/xCT/GPX4 pathway, which mediates the suppression of ferroptosis and lipid peroxidation, providing protection to the aorta.