| 摘要: |
| 目的 研究激活α7烟碱型乙酰胆碱受体 (α7 nicotinic acetylcholine receptor, α7nAchR) 对大脑皮质神经元氧糖剥夺损伤的作用及可能机制。方法 取体外培养7 d的原代大脑皮质神经元, 随机分为3组:对照组、氧糖剥夺组、PNU-282987(α7nAchR激动剂)组。氧糖剥夺组细胞氧糖剥夺12 h;PNU-282987组用PNU-282987预处理细胞24 h, 然后氧糖剥夺12 h。采用CCK-8测定3组大脑皮质神经元存活率, 比色法检测上清液乳酸脱氢酶 (lactate dehydrogenate, LDH) 含量, 流式细胞术检测细胞凋亡率和活性氧(reactive oxygen species, ROS)产量, 蛋白质印迹法检测血红素氧化酶 (hemeoxygenase-1, HO-1)、缺氧诱导因子1α (hypoxia inducible factor-1α, HIF-1α) 表达量。结果 与氧糖剥夺组相比, PNU-282987可提高大脑皮质神经元存活率(P<0.05), 降低LDH含量(P<0.05),抑制细胞凋亡(P<0.05), 降低ROS产量(P<0.05), 同时使HO-1蛋白表达升高、HIF-1α蛋白表达减少(P<0.05)。结论 激活α7nAchR具有抗大脑皮质神经元氧糖剥夺损伤作用, 该作用可能与抗氧化应激有关。 |
| 关键词: α7烟碱型乙酰胆碱受体 大脑皮质神经元 氧糖剥夺 氧化性应激 |
| DOI：10.3724/SP.J.1008.2015.00472 |
| 投稿时间：2015-01-13修订日期：2015-03-02 |
| 基金项目:上海市自然科学基金青年项目(13ZR145930010). |
|
| Activation of α7 nicotinic acetylcholine receptor alleviates cerebral cortical neuron injury induced by oxygen-glucose deprivation |
| XIN Ru-juan1,ZHAO Ting1,NI Min2,LI Dong-jie2,SHEN Fu-ming1* |
(1. Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai 200433, China;
2. Department of Clinical Pharmacy, the 10th People's Hospital of Shanghai, Tongji University, Shanghai 200072, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect of activating α7 nicotinic acetylcholine receptor (α7nAchR) on cerebral cortical neurons injury induced by oxygen-glucose deprivation (OGD) and the possible mechanism. Methods Cerebral cortical neurons cultured for 7 d were randomly divided into three groups: control group, OGD group (Cells experienced a 12 h oxygen-glucose deprivation) and OGD group treated with PNU-282987 (Cells experienced a 12 h oxygen-glucose deprivation with PNU-282987 pretreatment for 24 h). Cell viability was determined by CCK-8 assay, lactate dehydrogenase (LDH) was examined to reflect cell injury, apoptosis and reactive oxygen species (ROS) production were analyzed by flow cytometry, and expression of hemeoxygenase-1 (HO-1) and hypoxia inducible factor-1α (HIF-1α) were detected by Western blotting analysis. Results OGD resulted in cell death, LDH increase, and cell apoptosis. Compared with the OGD group, PNU-282987 pretreated group had significantly increased cell survival (P<0.05), significantly decreased LDH level and ROS production (P<0.05), and significantly inhibited cell apoptosis (P<0.05). Meanwhile, HO-1 protein expression was significantly increased and HIF-1α protein expression was significantly reduced in PNU-282987 pretreated group compared with the OGD group (P<0.05). Conclusion Activation of α7nAchR can protect cerebral cortical neurons against OGD-induced injury, which may be related to the anti-oxidative stress. |
| Key words: α7 nicotinic acetylcholine receptor cerebral cortical neurons oxygen-glucose deprivation oxidative stress |
