The Effects And Mechanisms Of Lipids On Neurons In Mouse Cerebral Ischemia-reperfusion Model

Research objective At present,stroke has become one of the diseases with high mortality and disability,which has a serious impact on the life and quality of life of human beings.However,the most effective treatment is the previous thrombolysis.The MCAO model of the animal stroke model,in this experiment,used to study the changes of lipids in the cerebral cortex damaged by cerebral ischemia/reperfusion in mice using the highly sensitive UPLC-MS mass spectrometry platform.Therefore,this experiment used lipidomics to find Lipid molecules of significant changes of brain ischemia-reperfusion injury in mice and those lipids effects on neuronal activity and molecular mechanisms of action were investigated.Research methods 1.Establish a mouse MCAO stroke model,and then use the ultra-high performance liquid chromatography to screen out the lipid molecules of significant differences.2.Take the fetal mice and the cerebral cortex.Use it as the primary neuron culture,and use immunofluorescence staining(MAP-2)to identify whether it is a neuron.3.The cultured primary neurons were seeded on 96-well plates.When the primary neurons were treated with the lipids screened in the early stage,Using CCK 8 kit to observe the effect of lipids on cultured neuronal viability.4.Several inhibitors of cell death and lipids were purchased to incubate primary neurons to measure cell viability by CCK 8 in order that explore the way cell death.5.The cultured primary neurons were seeded in a 35 mm culture dish.Then add different concentrations of lipids to detect intracellular calcium ion signal changes to explore whether lipids can cause neuronal excitability.Research results 1.Establish a mouse MCAO stroke model,then use the ultra-high performance liquid chromatography to quantitatively analyze the mouse cerebral cortex.A total of 578 lipids were screened,and then the differential lipids between the groups were analyzed.A total of 84 significant differences in lipid molecules were screened.2.Some significant changes in lipid molecules after stroke can significantly reduce the viability of primary neurons(lipid concentrations from 25 ?M to 50 ?M),leading to neuronal death.3.To explore the ways of lipid-induced neuronal death,it was found that some cell death inhibitors can significantly rescue the vitality of neurons,and MK-801 can also block the excitotoxicity caused by some lipids.Conclusion During the cerebral ischemia-reperfusion of mice,some significant changes in lipids play an important regulatory role in the recovery of stroke.During stroke recovery,some of these lipids,such as Phosphatidylcholine,Lysophosphatidylcholine,Phosp-hatidylethanolamine,Ceramides,Lysophosphatidylethanolamine can be induced microglia activation,next causing inflammatory responses and excitotoxicity of neurons,leading to neuronal death.