Butylphthalide Inhibits RIP3 In Necroptosis Pathway Of Neuronal Ischemia/reperfusion Injury In Vitro

Objective Through the construction of ischemia and hypoxia model by rat primary neurons,to observe the mechanism of butylphthalide(NBP)on rat primary neuron ischemia-reperfusion injury.To observe the effect of butylphthalide on the expression of RIP3 gene in neurons,explore the mechanism of Glyoxalase 1(GLO1)changes after necroptosis,and provide a new treatment strategy for ischemic stroke.Methods The oxygen-glucose deprivation(OGD)model was constructed using primary cultured rat cortical neurons in vitro.Neurons were divided into drug-treated group,OGD group and Control group,and the drug-treated group was treated with 10 ?mol/L butylphthalide or positive drug after OGD treatment.The degree of necrosis of neurons was measured using necrosis kit 48 hours after ischemia-reperfusion.The expression of RIP3 mRNA in cells was measured at different time points after reperfusion(3h/6h/12h/24h/48h).The activity of different groups of GLO 1 was detected using ELISA kit.Results The neuron necrosis rate of the OGD group was significantly higher than that of the Control group.The neuron necrosis after butylphthalide intervention was significantly lower than that of the model control group(P<0.05).The mRNA of RIP3 at each time point of pretreatment of neurons with butylphthalide was significantly lower than that of the OGD group(P<0.05).The OGD group down-regulated the expression of GLO1,while the butylphthalide pretreatment increased GLO1 activity(P<0.05).Conclusion Over-activated RIP3 inhibits GLO1 activity after neuronal ischemia-reperfusion injury,while butylphthalide can inhibit RIP3 expression and restore GLO1 activity.Butylphthalide may regulate neuronal necrosis by inhibiting the expression of RIP3.