引用本文 信如娟, 赵婷, 倪敏, 李冬洁, 沈甫明. 激活α7烟碱型乙酰胆碱受体减轻大脑皮质神经元氧糖剥夺损伤[J]. 第二军医大学学报, 2015, 35(5): 472-476 XIN Ru-juan, ZHAO Ting, NI Min, LI Dong-jie, SHEN Fu-ming. Activation of α7 nicotinic acetylcholine receptor alleviates cerebral cortical neuron injury induced by oxygen-glucose deprivation[J]. Acta Petrologica Sinica, 2015, 35(5): 472-476 (in Chinese with English abstract)

激活α7烟碱型乙酰胆碱受体减轻大脑皮质神经元氧糖剥夺损伤

1 , 赵婷1, 倪敏2, 李冬洁2, 沈甫明1 信如娟, 赵婷, 倪敏, 李冬洁, 沈甫明

1. 第二军医大学药学院药理学教研室, 上海 200433;

2. 同济大学附属上海第十人民医院药学部, 上海 200072 基金项目：上海市自然科学基金青年项目(13ZR145930010). 作者简介：信如娟,硕士生.E-mail:xinrujuan1112@126.com 通信作者(Corresponding author)：沈甫明,Tel:021-81871276,E-mail:fumingshen@126.com

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

Activation of α7 nicotinic acetylcholine receptor alleviates cerebral cortical neuron injury induced by oxygen-glucose deprivation

1 , ZHAO Ting1, NI Min2, LI Dong-jie2, SHEN Fu-ming1 XIN Ru-juan, ZHAO Ting, NI Min, LI Dong-jie, SHEN Fu-ming

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

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