RIP1/3对创伤性脑损伤后神经元的作用及其机制研究

摘要

目的 探讨RIP1/3对创伤性脑损伤(TBI)后神经元的影响及其作用机制.方法 将体外培养的皮层神经元分为4组:siRIP组、Nec-1预处理组、阴性质粒转染组和对照组.通过慢病毒转染法分别干涉RIP1、RIP3表达,建立离体TBI损伤模型,通过流式细胞学检测划伤后神经元存活情况.对体外培养的皮层神经元敲除RIP1/3,建立谷氨酸诱导神经元损伤模型,通过流式细胞学检测谷氨酸刺激后神经元存活情况.结果 siRIP组及Nec-1预处理组机械性损伤后神经元存活率高于阴性质粒转染组和对照组.Nec-1预处理组谷氨酸损伤后神经元存活率高于对照组,而siRIP组存活率与对照组和阴性转染组比较未见显著差异.结论 RIP1和RIP3可能对TBI诱导后神经元死亡有作用,而RIP1抑制剂Nec-1可能对TBI具有脑保护作用.RIP1/3与谷氨酸兴奋毒性诱导细胞死亡无关,而Nec-1对于谷氨酸损伤具有潜在保护作用,并可能存在特异性靶点.%Objective To investigate the effect and mechanism of RIP1/3 on neurons after traumatic brain injury (TBI).Methods The cultured cortical neurons were divided into four groups.:siRIP group,Nec-1 pretreatment group,negative plasmid transfection group and control group.The TBI damage model was established by interfering with the expression of RIP1 and RIP3 through lentiviral transfection,respectively.The RIP3 damage model was established,and the survival rate of neurons was detected by flow cytometry.Results After mechanical injury,the survival rate of neurons in the siRIP group and in the Nec-1 pretreatment group was higher than that in the negative plasmid transfection group and in the control group.After glutamate stimulation,the survival rate of neurons in the Nee-1 pretreatment group was higher than that in the control group.siRIP3 group or siRIP1 group showed no significant difference in the survival rate of neurons when contrasted withthe negative control transfection group.Conclusion RIP1 and RIP3 may play a role in neuronal death induced by TBI,RIP1 inhibitor Nec-1 may have a function in brain protection for TBI.RIP1 and RIP3 have no effect on neuron cell death induced by glutamate excitotoxicity.Nec-1 has a potential protective effect on glutamate injury,probably suggesting a specific target.