缺氧缺血新生大鼠神经元自噬基因表达节律及调控机制

发布时间：2018-06-15 07:24

  本文选题：缺氧缺血 + 自噬　； 参考：《中国当代儿科杂志》2017年08期

【摘要】：目的通过新生大鼠缺氧缺血脑损伤后神经元自噬基因和节律基因的表达,探讨缺氧缺血引起神经损伤的新机制。方法将12只Sprague-Dawley大鼠随机分为缺氧缺血组和假手术组,每组6只。采用结扎并切断大鼠右侧颈总动脉并给予低氧处理的方法建立体内缺氧缺血脑损伤模型。Western blot检测两组大鼠大脑皮层和海马组织节律蛋白Clock表达情况。体外培养大鼠神经元细胞,随机分为氧糖剥夺(OGD)组和对照组,OGD组加入无糖无血清DMEM培养基模拟细胞缺血状态,并给予低氧处理建立体外缺氧缺血脑损伤模型。采用Western blot检测两组不同时间点自噬相关蛋白Beclin1和LC3,以及节律基因Clock蛋白表达情况。应用si RNA技术抑制神经元Clock蛋白表达后,检测Beclin1和LC3的表达变化。结果体外培养神经元Beclin1和LC3Ⅱ的表达呈现节律性波动;OGD处理后,体外培养神经元Beclin1和LC3Ⅱ的表达随着时间的延长逐渐升高,不再呈现节律性波动;与假手术组相比,缺氧缺血引起大鼠皮层和海马组织Clock表达降低(P0.05);体外培养神经元经OGD处理后,Clock表达也较对照组显著降低(P0.05);与阴性对照组相比,抑制神经元节律基因Clock表达后,自噬相关蛋白Beclin1和LC3Ⅱ的表达均显著下降(P0.05)。结论缺氧缺血引起神经元Beclin1和LC3Ⅱ表达节律紊乱,其机制可能与Clock参与调控Beclin1和LC3Ⅱ的表达有关。
[Abstract]:Objective to investigate the expression of autophagy gene and rhythm gene after hypoxic-ischemic brain injury in neonatal rats and to explore the new mechanism of hypoxia-ischemia induced neuronal injury. Methods Twelve Sprague-Dawley rats were randomly divided into hypoxic-ischemic group and sham-operation group with 6 rats in each group. The rat model of hypoxic-ischemic brain injury was established by ligating and cutting off the right common carotid artery and treated with hypoxia. Western blot was used to detect the expression of circadian rhythm protein clock in cerebral cortex and hippocampus of rats in both groups. Rat neuronal cells were cultured in vitro and randomly divided into two groups: the OGD group and the control group. The model of hypoxic-ischemic brain injury was established by hypoxia-induced hypoxia in the control group (OGD group) and the control group (OGD group) by adding the glucose free serum-free DMEM medium to simulate the ischemic state of the cells. The expression of Beclin1 and LC3 and clock protein were detected by Western blot at different time points. The expression of Beclin1 and LC3 was detected by si RNA technique after inhibiting the expression of clock protein in neurons. Results the expression of Beclin1 and LC3 鈪，

本文编号：2021206