Sofren Injection Protects Against Ferroptosis In HT22 Cells After Oxygen-glucose Deprivation/Reoxygenation Based On Nrf2/HO-1 Signaling Pathway

BackgroundAfter cardiopulmonary resuscitation(CPR),Ischemia/reperfusion injury(I/R)will lead to obvious neurological impairment,patients often have a poor prognosis.Therefore,early intervention to improve the anti-hypoxia ability of neurons is of great significance to improve the level of cardiopulmonary resuscitation and improve the quality of life of patients.There have been a lot of researches over the years,but little effect has been achieved.Ferroptosis is a hot topic of research on cerebralischemia reperfusioninjury(CIRI).The occurrence of ferroptosis is mainly regulated by three pathways,including antioxidant pathway,iron metabolism pathway and ROS metabolism pathway.GPX4 is a key gene in the antioxidant pathway.Multiple studies have shown that GPX4 is the only gene mediated by the Nrf2 transcriptional pathway.Based on the severe oxidative stress injury and cell apoptosis after cerebral ischemia/reperfusion injury,and the important role of Nrf2/HO-1 signaling pathway in anti-oxidative stress injury and cell apoptosis,more and more studies have focused on the role of Nrf2/HO-1 signaling pathway in cerebral ischemia/reperfusion injury.Rhodiola Rosa L.(Rho)has beneficial effects of warming Yang,strengthening positive body and removing body,and its effect on cerebral ischemia reperfusion injury has been paid more and more attention in recent years,but its clinical application has not been effectively promoted.This study aims to clarify the ameliorating effect of Rho in CIRI,provide data support for Rho as an anti-brain injury after cardiopulmonary resuscitation,and clarify its mechanism of action.MethodIn this study,HT22 was used as the research object.Oxygen-glucose deprivation/reoxyge-nation(OGD/R)model and Erastin induced ferroptosis model were used.Ferrostatin-1(FER-1)was used as the positive control.The details are as follows:Experiments in vitro:The hippocampal neuronal cell line(HT22)were divided into:(1)Control group(2)OGD/R group(3)OGD/R+Rho low-dose group(4)OGD/R+Rho high-dose group(5)OGD/R+Fer-1 group(6)Erastin group(7)Erastin+Rho low-dose group(8)Erastin+Rho high-dose group(9)Erastin+Fer-1 group,cell morphological changes were observed under the microscope,cell survival rate was detected by CCK-8 method,the relative quantification of iron-dead cells by C11 Bodipy 581/591 staining,the nucleation rate of Nrf2 and the fluorescence intensity of HO-1 were detected by immunofluorescence staining.The expressions of Nrf2,HO-1,FACL4,Tf,COX2 and GPX4 were determined by Western Blot.ResultsIn the OGD/R model of HT22 hippocampal neurons in mice,Sofren injection could reduce ferroptosis,and Ferrostatin-1,an ferroptosis inhibitor,could also have the same effect.In the HT22 model of ferroptosis in hippocampal neurons,20 ml/1 Sofren injection was effective in reducing ferroptosis in cells,which was not significantly different from Ferrostatin-1.In the HT22 hippocampal neuron OGD/R model and iron death model,Sofren injection can increase the nucleation rate of Nrf2,up-regulate the expressions of HO-1,GPX4 and FACL4,down-regulate the expressions of Tf and COX2,thereby reducing ferroptosis of cells.There was no significant difference compared with Ferrostatin-1 positive controls.ConclusionSofren injection can inhibit ferroptosis through Nrf2/HO-1 pathway and improve hypoxia and reoxygenation injury of HT22.