Notch/JNK信号在低氧致大鼠皮层神经元损伤中的作用研究

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

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。

Objective： To study the imvolvment of Notchl/JNK signaling pathway in neuron injury caused by hypoxia. Methods： The primary cortical neurons were cuhured and hypoxia model was setted up using hypoxia conditions. The via- bility 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 Notchl 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 condi- tions, 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 Caspase- 3, there by reducing the viability of neurons or promoting neuronal death.