安徽医科大学学报2025，Vol.60Issue(1)：31-40,10.DOI:10.19405/j.cnki.issn1000-1492.2025.01.005
参附注射液通过抑制铁死亡发挥对缺氧缺血性脑损伤的保护作用
Protective effect of Shenfu injection against neonatal hypoxic-ischemic brain injury by inhibiting the ferroptosis
摘要
Abstract
Objective To observe the brain tissue injury during hypoxia-ischemia,as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI),and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD)by inhibiting ferroptosis.Methods An animal model of HIBD in SD rats was constructed and intervened with SFI.Pathologic changes in brain tissue were ob-served by HE staining methods.Nissen staining was used to observe neuron survival.Glutathione Peroxidase 4(GPX4)and Divalent Metal Transporter 1(DMT1)expression were detected in brain tissue by Western blot,im-munohistochemistry and immunofluorescence.Reduced Glutathione(GSH),Lactate Dehydrogenase(LDH),Ma-londialdehyde(MDA),Superoxide Dismutase(SOD)and tissue iron content were determined with the kits.BV-2 microglial cell line(BV2)cells were cultured in vitro and divided into control group(Ctrl group),oxygen-glucose deprivation group(OGD group),iron ferroptosis-inducing group(Erastin group),iron ferroptosis-inhibiting group(Fer-1 group),Shenfu injection group(SFI group),and Erastin+Shenfu injection group(Erastin+SFI group).2',7'-Dichlorodihydrofluorescein diacetate(DCFH-DA)reactive oxygen species(ROS)fluorescent probe was used to detect the ROS release level;Immunofluorescence was used to observe intracellular GPX4,DMT1 expression.Results Compared with the Sham group,rats in the HIBD group showed significant neuronal cell damage in brain tissue,decreased GPX4 expression(P＜0.01),increased DMT1 expression(P＜0.01),decreased GSH and SOD levels(P＜0.01),and increased LDH,MDA and tissue iron levels(P＜0.05,P＜0.05,P＜0.01).In contrast,after the intervention of SFI,GPX4 expression was elevated(P＜0.01),DMT1 expression decreased(P＜0.01),GSH and SOD levels were elevated(P＜0.01),and LDH,MDA,and tissue iron levels decreased(P＜0.05,P＜0.05,P＜0.01).The cells experiments showed that compared with the Ctrl group,the OGD group had a signifi-cantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity,and an increase in the fluorescence intensity of DMT1(P＜0.01),compared with the OGD group,the ROS content was reduced in the SFI group,while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P＜0.01).Con-clusion Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats,and their brain tis-sues show decreased expression of GPX4 and increased expression of DMT1.The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats.In contrast,Shenfu injection has a protective effect on HIBD experi-mental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.关键词
缺氧缺血性脑损伤/铁死亡/氧化应激/氧糖剥夺/参附注射液/小胶质细胞Key words
hypoxic-ischemic brain damage/ferroptosis/oxidative stress/oxygen glucose deprivation/Shenfu in-jection/BV2 cells分类
医药卫生引用本文复制引用
张晓彤,张萌,李刚,胡阳,荀雅静,丁会,沈栋林,吴铭..参附注射液通过抑制铁死亡发挥对缺氧缺血性脑损伤的保护作用[J].安徽医科大学学报,2025,60(1):31-40,10.基金项目
江苏省卫生健康委员会科研项目(编号:H2019004) （编号:H2019004）
江苏省研究生科研与实践创新计划(编号:KYCX22_2911) （编号:KYCX22_2911）
江苏省大学生创新创业计划(编号:202010313048Y) Scientific Research Project of Jiangsu Health Commission(No.H2019004) （编号:202010313048Y）
Graduate Student Research and Innovation Program of Jiangsu Province(No.KYCX22_2911) （No.KYCX22_2911）
College Student Innovation and En-trepreneurship Training Project of Jiangsu Province(No.202010313048Y) （No.202010313048Y）
