Role Of Receptor-interacting Protein 3 And Classical Necroptosis Signaling Pathway In Cerebral Ischemia-reperfusion (I/R) Injury

[Background] Cell death involves multiple aspects of life, the positive effects:embryonic development, cells of the body’s " metabolism", the negative effects:inflammation, cancer and so on. Past view:Cell death except Apoptosis is Active Deathmatch, accompany with Signaling molecules associated and energy consumption eventually leading to cell death by gene。 2005 Degterev put forward a novel cell death:necroptosis.This way of cell death like structure having characteristics of membrane integrity damage surrounding tissue inflammation, but the process is subject to a series of intracellular signaling molecules regulating. Cerebral ischemia induced brain damage is one of the main causes of disability and death in Neurology. Through in-depth study of the biochemical mechanism of necroptosis pathway proven its role in various disease models, not only beneficial to the necroptosis way to deepen understanding and awareness, but also to a neurologist ischemia-reperfusion injury related diseases therapeutic targets Research confirmed and treatment drugs is important.[Purpose] To study variation in vitro neuronal cells after ischemia-reperfusion injury of RIP3. The study inspired by cerebral ischemic penumbra area after clinical appearance.To study and explore the necroptosis distributed in ischemic penumbra area, furtherly reveal necroptosis Mechanism in cerebral ischemia-reperfusion injury in rats.[Methods] Primary neurons culture, neuronal cells fluorescent identification, RIP3 fluorescent identification, HT22 cells culture and cryopreservation, cell Hypoxia model Manufacture, animals (I/R) injury model Manufacture, PCR technology, Western Blotting technique, MTT technology, mouse TTC staining brain[Results] expression in ischemia-reperfusion in vivo and in vitro models have changed significantly. In this experiment, the set point, the 12 hours after RIP3 expression in ischemia-reperfusion injury peaked.Consistent source of mouse cell line HT22 hippocampal and cortical neurons in rats after modeling PCR results same trend. The animal modeling infarct zone and ischemic penumbra area Western Blotting consistent same trend, consistent with cell PCR trend. In vivo, the infarct vs the penumbra area, RIP3 expression fluctuate significantly.1 hour hypoxia NEC-1 in the experimental group than in the control group increased cell survival is not obvious, and hypoxia for 4 hours was significantly higher in the experimental group NEC-1 cell viability than the control group.[Conclusions] Cerebral ischemia-reperfusion model, infarct zone and ischemic penumbra are different due to varying degrees of damage, necroptosis participation in infarct zone is more active.