Protective effect of Shenfu injection against neonatal hypoxic-ischemic brain injury by inhibiting the ferroptosis
10.19405/j.cnki.issn1000-1492.2025.01.005
- Author:
Xiaotong Zhang
1
;
Meng Zhang
1
;
Gang Li
2
;
Yang Hu
3
;
Yajing Xun
3
;
Hui Ding
1
;
Donglin Shen
4
;
Ming Wu
5
Author Information
1. The First Clinical Medical College，Xuzhou Medical University，Xuzhou 221000
2. Dept of Stomatology，The Affiliated Hospital of Xuzhou Medical University，Xuzhou 221000
3. The Second Clinical Medical College， Xuzhou Medical University，Xuzhou 221000
4. Dept of Pediatrics，The Affiliated Hospital of Xuzhou Medical University，Xuzhou 221000
5. The First Clinical Medical College，Xuzhou Medical University，Xuzhou 221000; Dept of Pediatrics，The Affiliated Hospital of Xuzhou Medical University，Xuzhou 221000
- Publication Type:Journal Article
- Keywords:
hypoxic-ischemic brain damage;
ferroptosis;
oxidative stress;
oxygen glucose deprivation;
Shenfu in- jection;
BV2 cells
- From:
Acta Universitatis Medicinalis Anhui
2025;60(1):31-40
- CountryChina
- Language:Chinese
-
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 observed 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, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand 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 significantly 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).
Conclusion :Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues 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 experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.
- Full text:2025030416180237677参附注射液通过抑制铁死亡发...缺氧缺血性脑损伤的保护作用_张晓彤.pdf
