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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 investigate the role of Notch1 / JNK signaling pathway in hypoxic-induced neuronal injury. Methods: Primary cultured neonatal SD rat cortical neurons were cultured under hypoxic conditions to establish hypoxia model. The activity of neurons and the activity of neurons after siRNA interference were detected by CCK-8 assay at different times of hypoxia. The expression of NICD and p-JNK in control group and hypoxia group were detected by RT-q PCR and Western Blot Caspase-3mRNA and Caspase-3mRNA were detected by Western Blot after transfection of Notch1 siRNA or JNK siRNA into neurons by siRNA transfection. 3 protein expression. Results: Under hypoxia, the activity of neurons was low and time-dependent. In hypoxia, the expression of NICD, p-JNK, Caspase-3 mRNA and protein in neurons were significantly increased. Inhibition of Notch1 neurons in the activity, can further inhibit the expression of JNK, Caspase-3, improve neuronal activity. CONCLUSION: Hypoxia leads to the activation of Notch1 signal in neurons, Notch signaling to regulate the phosphorylation of JNK, and phosphorylation of JNK further promotes the activation of Caspase-3, thereby decreasing the activity of nerve cells or promoting the death of nerve cells.