摘 要：为探讨丙泊酚（Pro）调控沉默信息调节因子1（SIRT1）/p53信号通路介导的铁死亡对H9C2心肌细胞缺氧/复氧（H/R）损伤的影响，构建体外H9C2细胞H/R模型，分为对照组、H/R组、H/R+Pro组、H/R+Pro+erastin组、
H/R+Pro+si-NC组和H/R+Pro+si-SIRT1组。用细胞计数试剂盒-8（CCK-8）检测Pro对H9C2细胞的毒性作用；用2', 7'-二氯二氢荧光素二乙酸酯（DCFH-DA）荧光探针染色检测各组细胞活性氧（ROS）的产生水平；用透射电子显微镜检测线粒体的超微结构；用相应的试剂盒检测氧化应激指标；用蛋白质免疫印迹法检测铁死亡相关蛋白的表达。结果显示，与对照组相比，H/R组的细胞活力降低，ROS和丙二醛（MDA）水平升高，过氧化氢酶（CAT）和超氧化物歧化酶（SOD）水平减少，核膜破裂，线粒体膜密度增加，线粒体嵴减少，出现肿胀、断裂的情况，Fe2+含量、乳酸脱氢酶（LDH）活性增加，长链脂酰辅酶A合成酶4（ACSL4）和p53水平升高，谷胱甘肽过氧化物酶4（GPX4）和SIRT1水平降低（P<0.05）。Pro处理使得H/R损伤得到显著改善，细胞活力提升，Fe2+含量增加，ROS、MDA、ACSL4、p53水平降低，SOD、CAT、GPX4、SIRT1水平升高，线粒体结构修复（P<0.05）。erastin可部分逆转Pro的保护作用（P<0.05）。si-SIRT1可完全阻断Pro效应，使GPX4水平降低，Fe2+含量增加，ACSL4水平升高（P<0.05）。本研究揭示了Pro可能通过激活SIRT1/p53通路抑制铁死亡，进而对心肌细胞H/R损伤发挥保护作用。该发现为Pro实现心肌保护提供了关键机制依据，为防治心肌缺血再灌注损伤提供了潜在的新策略。
关键词：丙泊酚；沉默信息调节因子1/p53信号通路；铁死亡；心肌细胞；缺氧/复氧损伤
中图分类号：R542.2+2      文献标志码：A                   DOI：10.3969/j.issn.1007-7146.2025.06.010

Abstract: To investigate the effects of propofol (Pro) on hypoxia/reoxygenation (H/R) injury in H9C2 cardiomyocytes via regulation of the silent information regulator 1(SIRT1)/p53 signaling pathway-mediated ferroptosis, an in vitro H9C2 cell H/R model was established. Cells were divided into the following groups: Control group, H/R group, H/R+Pro group, H/R+Pro+erastin group, H/R+Pro+si-NC group, and H/R+Pro+si-SIRT1 group. The cytotoxic effect of Pro on H9C2 cells was assessed using the cell counting kit-8 (CCK-8) assay. Intracellular reactive oxygen species (ROS) levels were measured using the 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe. Mitochondrial ultrastructure was examined by transmission electron microscopy (TEM). Oxidative stress markers were quantified using corresponding assay kits. Expression levels of ferroptosis-related proteins were detected by Western blot. Results demonstrated that compared to the Control group, the H/R group exhibited significantly reduced cell viability, elevated levels of ROS and malondialdehyde (MDA), decreased levels of catalase (CAT) and superoxide dismutase (SOD), nuclear membrane rupture, increased mitochondrial membrane density, loss of mitochondrial cristae, swelling and rupture of mitochondria, increased Fe2+ content and lactate dehydrogenase (LDH) activity, elevated levels of acyl-CoA synthetase long-chain family member 4 (ACSL4) and p53, and reduced levels of glutathione peroxidase 4 (GPX4) and SIRT1 (P<0.05). Pro treatment significantly ameliorated H/R injury, increasing cell viability and elevating SOD, CAT, GPX4, and SIRT1 levels, while reducing Fe2+ content, ROS, MDA, ACSL4, and p53 levels, and restoring mitochondrial structure (P<0.05). Erastin partially reversed the protective effects of Pro (P<0.05). Conversely, si-SIRT1 completely abolished the effects of Pro, resulting in decreased GPX4 levels and increased Fe2+ content and ACSL4 levels (P<0.05). This study revealed that Propofol likely protects cardiomyocytes against H/R injury by activating the SIRT1/p53 pathway to inhibit ferroptosis. This finding provides crucial mechanistic insight into Propofol-mediated cardioprotection and suggests a potential novel strategy for the prevention and treatment of myocardial ischemia-reperfusion injury.
Key words: propofol; silent information regulator 1/p53 signaling pathway; ferroptosis; cardiomyocytes; hypoxia/reoxygenation injury
（Acta Laser Biology Sinica, 2025, 34(6): 561-568）