Notch/JNK信号在低氧致大鼠皮层神经元损伤中的作用研究

目的:探讨Notch1/JNK信号通路在缺氧引起的神经元损伤中的作用。方法:原代培养新生SD大鼠皮层神经元,利用低氧条件培养建立缺氧模型。利用CCK-8实验检测在低氧诱导的不同时间,神经元的活性及siRNA干扰后神经元的活性;利用RT-q PCR与Western Blot检测对照组与低氧组神经元中NICD,p-JNK,Caspase-3mRNA与蛋白的表达情况;利用siRNA转染技术转染Notch1 siRNA或JNK siRNA至神经元中后,利用Western Blot检测低氧组与转染组神经元中NICD,p-JNK,Caspase-3蛋白的表达情况。结果:缺氧情况下神经元活性较低,具有时间依赖性;缺氧情况下,神经元的NICD,p-JNK,Caspase-3 mRNA及蛋白的表达均显著升高。抑制神经元中Notch1的活性,可进一步抑制JNK,Caspase-3的表达,改善神经元活性。结论:缺氧导致神经元Notch1信号被激活,Notch信号调节JNK的磷酸化,磷酸化的JNK进一步促进Caspase-3的激活,从而降低神经细胞的活性或促进神经细胞的死亡。

The imvolvment of Notch/JNK signaling pathways in the rat cerebral neuronal injury induced by hypoxia

Objective:To study the imvolvment of Notch1/JNK signaling pathway in neuron injury caused by hypoxia.Methods:The primary cortical neurons were cultured and hypoxia model was setted up using hypoxia conditions.The viability of neurons was detected at different time by CCK-8 assay.The expression of NICD,p-JNK and Caspase-3 mRNA and protein in neurons of hypoxia group and control group was detected by RT-qPCR and Western Blot.Using siRNA transfection technique to transfect Notch1 siRNA or JNK siRNA into neurons,then the expression of NICD,p-JNK and Caspase-3 protein in neurons of hypoxia group and siRNA group was detected by Western Blot.Results:The viability of neurons in hypoxia was low,and the viability of neurons decreased gradually with time dependence.Under hypoxia conditions,the expression of NICD,p-JNK,Caspase-3 mRNA and protein increased,but inhibition Notchl could further inhibit the expression of JNK and Caspase-3,inhibited the activity of neuron.Conclusion:Under hypoxia condition,the activation of Notch signaling pathways might regulate phosphorylation of JNK,and then imduce the activation of Caspase3,there by reducing the viability of neurons or promoting neuronal death.