Propofol Reduces Ferroptosis In Glutamate-and Erastin-induced Neuron Injury

Background: With the development of the aging society,the incidence of aging diseases is getting higher and higher,which also brings huge burden and loss to the society and families.The pathological mechanisms of neurodegenerative diseases are complex,which including oxidative stress,apoptosis of neuron,and excitatory toxicity of glutamate.Ferroptosis is a newly form of regulatory necrosis characterized by intracellular iron ion accumulation and lipid oxidation.In recent years,many studies have shown that ferroptosis and neurodegenerative diseases are closely related.Propofol is a short-acting intravenous anesthetic commonly used in clinical practice.Propofol not only has anesthetic effect,but also neuroprotective effect.Therefore,this experiment used glutamate toxicity model to simulate the excitability toxicity of neurodegenerative disease factors,using ferroptosis specificity inducers(erastin)to simulate the ferroptosis of neurodegenerative disease caused by factors,explore propofol in neurodegenerative disease excitability toxicity factors and influences of die factors of ferroptosis.Methods: HT22 cells was selected randomly,using propofol pretreatment for 2 hours,respectively,treat with 5 m M glutamate and 0.5 μM Erastin for eight hours,we observed the cell morphology,measured cell survival rate,intracellular reactive oxygen species,intracellular lipid oxidation,intracellular iron ion change situation,and PTGS2(specific markers of ferroptosis)transcription level change.Western blot was used to detect the levels of p-erk,ERK,p(S663)-ALOX5 and ALOX5 protein levels.Next,we transfected plasmids which encodes WT-ALOX5,S663A-ALOX5 to build transient overexpression cell lines.p-ERK,ERK,p(S663)-ALOX5,ALOX5 protein levels,cell viability,intracellular lipid oxidation levels,and intracellular iron ion changes were detected after 8 hours of treatment with 0.25μM Erastin,respectively.To compare the protein levels of p-erk,ERK,p(S663)-alox5 and ALOX5 between WT-ALOX5 and S663A-ALOX5,as well as the cell survival rate,intracellular lipid oxidation level and intracellular iron ion change level.Results: compared with the glutamate and Erastin groups,the cell survival rate was significiantly increased and intracellular reactive oxygen species,lipid oxidation and iron ion levels in propofol group were significantly decreased.The expression level of ferroptosis marker PTGS2 decreased significantly in the propofol group compared with the Erastin group,while no significant decrease occurred in the propofol group compared with the glutamate group.The ratio of p-erk /ERK protein levels in the propofol group showed no significant change compared with the glutamate group and Erastin group,while the ratio of p-ALOX5 /ALOX5 protein levels in the propofol group was significantly lower than that in the Erastin group.After transfection of overexpressed plasmids encodes WT-ALOX5 and S663A-ALOX5,the cells transfected with S663A-ALOX5 lost the inhibitory effect of propofol on the ferroptosis of Erastin compared with those transfected with WT-ALOX5.Conclusion: Propofol can significantly inhibit the neurotoxicity of glutamate and Erastin,and reverse the rise of intracellular ROS,intracellular lipid oxidation and iron ion accumulation caused by glutamate and Erastin,and inhibit ferroptosis caused by glutamate and Erastin.After overexpression of WT-ALOX5 and S663A-ALOX5,propofol was not able to eliminate ferroptosis caused by Erastin in S663A-ALOX5 transient transfected cells compared with WT-ALOX5.