Establishment Of The Cerebral Ischemia And Reperfusion Model And Study On Protective Effect Of C3a C5a Receptor Inhibition After Murine Ischemic-reperfusioninjury

Part one：Establishment of the cerebral ischemia and reperfusionmodelObjective：To establish stable and repeatable focal cerebral ischemia reperfusion modelmice．Methods：Male, ripe C57Bl/6mice were selected to establish the instant middle cerebralartery occlusion (MCAO) model by intraluminal occlusion using monofilament. The timesof MCAO were60min respectively.24h after operation, we assessed nervous functiongrades of mice, and observed the reliability of the models.Results: Model group showed nervous function obstacle and TTC staining clearlyindicated the brain infarct.Conclusion: We successfully established cerebral ischemia-reperfusion mice model, andthey are suitable for ischemia-reperfusion injury study. Part two：Study on protective effect of C3a C5a receptor inhibitionafter murine ischemic-reperfusion injuryObjective: In this study, we inhibite the C3a, C5a receptor level of activity to investigatethe role of C3a, C5a receptor in ischemia-reperfusion injury. Explore C3aR, C5aR inhibitorin cerebral ischemia and reperfusion injury in mice.Methods: C3a and C5a receptor antagonist or vehicle were administered to mice beforetemporary middle cerebral artery occlusion.24h after operation, we assessed the results ofischemia-reperfusion injury，and TTC staining of2mm brain slice was used to calculatedthe infarct volumes.Results: Animals treated with C3a and C5a receptor antagonist experienced significantlydecreased infarct volumes and demonstrated an improving trend in neurological function.Conclusion: These findings demonstrate that modulation of C3a and C5a receptor activitysignificantly alters the degree of neurological damage after experimental reperfused stroke. Part three：Study on protective effect of C3a C5a receptor defects inmurine ischemic-reperfusion injuryObjective: In this study, through the making C3a, C5a receptor-deficient mice with focalcerebral ischemia reperfusion model, we explore C3a, C5a in cerebral ischemia andreperfusion injury in mice.Methods: Five groups were divided and they are normal BALB/c mice group, shamoperation group, normal BALB/c mice PBS group, C3aR-deficient mice group andC5aR-deficient mice group. Every group have6mice. Temporary occlusion the middlecerebral artery of mice.24h after operation, we assess the result of ischemia-reperfusioninjury，and nervous function grades of mice were carried out. TTC staining of2mm brainslice was used to calculate the infarct volumes.Results: C3aR-deficient mice group, C5aR-deficient mice group, neurological functionhas improved, significant reduction in infarct volume.Conclusion: The study results show the C3aR, C5aR defects group miceischemia-reperfusion injury severity significantly improved.