Protective mechanism of Yiqi Jiedu formula against ferroptosis in cerebral ischemia via GPX4 pathway
Objective To determine the protective mechanism of Yiqi Jiedu formula(YQ)against ferroptosis in acute ischemic stroke.Methods The rat model of acute cerebral ischemia was established by middle cerebral artery occlusion(MCAO).The PC12 cell glucose and oxygen deprivation(OGD)model was also established.The neurobehavioral scores and infarct volume were measured 24 h after the ischemia.The content of short-chain fatty acids(SCFA)in the colonic content of MCAO rats was detected by ELISA.The Fe2+in the ischemic brain tissue was observed by Prussian blue staining.The mitochondrial membrane potential and activity in the ischemic brain tissue were detected by JC-1 and resazurin,respectively.The content of ferrous ion,glutathione(GSH),malondialdehyde(MDA),superoxide dismutase(SOD)in the ischemic brain tissue and triglyceride(TG),high density lipoprotein cholesterol(HDL-C),and low density lipoprotein cholesterol(LDL-C)in the serum were measured.Molecular docking was used to predict YQ binding to the GPX4 protein.Western blot was used to detect the protein expression levels of G protein-coupled receptor 41(GPR41),glutathione peroxidase 4(GPX4)and p53 in the ischemic brain tissue of rats.Flow cytometry and Western blot were used to detect the protein expression level of GPX4 in the cells.Results Compared with the model group,the neurobehavioral scores and cerebral infarction volume of the middle and high dose YQ groups were decreased(P＜0.01),SCFA content in colon contents was increased(P＜0.01),iron particle deposition in the ischemic brain tissue was decreased,and the mitochondrial membrane potential increased(P＜0.05),while the mitochondrial viability was increased(P＜0.01).The levels of GSH,SOD,GPR41 and GPX4 were increased(P＜0.05 or P＜0.01),and those of Fe2+,MDA and p53 in the ischemic brain tissue were decreased(P＜0.01).The serum TG and LDL-C were significantly decreased(P＜0.05,P＜0.01),while HDL-C increased(P＜0.05).Compared with the OGD group,the expression of GPX4 protein in both the YQ group and Fer-1 group was increased(P＜0.01).Conclusion YQ can inhibit ferroptosis,another key mechanism for its protective effect against acute cerebral ischemia,whose action may be closely related to the GPX4-related pathway.
万方数据
维普
