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浙江大学学报(医学版)  2014, Vol. 43 Issue (3): 257-264    DOI: 10.3785/j.issn.10089292.2014.05.001
专题报道     
孟鲁司特和HAMI 3379对体外大鼠皮层神经元
徐栋敏1,张霞燕1,王晓蓉1,陈璐2,张丽慧2,石巧娟1,方三华1,卢韵碧1,张纬萍1,魏尔清1
1.浙江大学医学院药理学系,浙江 杭州 310058
2.杭州师范大学基础医学部药理学教研室 杭州市医学神经生物学重点实验室,浙江 杭州 310036
Antioxidative effects of cysteinyl leukotriene receptor antagonists montelukast and HAMI 3379 on ischemic injury in rat cortical neurons in vitro
XU Dong-min1, ZHANG Xia-yan1, WANG Xiao-rong1, CHEN Lu2, ZHANG Li-hui2, SHI Qiao-juan1, FANG San-hua1, LU Yun-bi1, ZHANG Wei-ping1, WEI Er-qing1
1 Department of Pharmacology, Zhejiang University School of Medicine, Hangzhou 310058, China;2Department of Pharmacology, School of Basic Medicine, Hangzhou Normal University; Hangzhou Key Laboratory of Neurobiology, Hangzhou 310036, China
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摘要: 

目的:在体外培养大鼠皮层神经元缺血性损伤模型中探讨两种半胱氨酰白三烯受体(CysLT1R和CysLT2R)拮抗剂——孟鲁司特和HAMI 3379的抗氧化作用。方法:以CysLT1R拮抗剂孟鲁司特和CysLT2R拮抗剂HAMI 3379预处理后,对大鼠皮层神经元进行缺糖缺氧/恢复或过氧化氢处理,观察神经元活性氧产生、线粒体膜电位、神经元存活率变化以及两种药物的作用。同时,以RNA干扰抑制CysLT1R和CysLT2R表达,观察以上变化。结果:缺糖缺氧处理1 h后神经元活性氧产生增加(2700%±68%,P<001),在缺糖缺氧恢复30 min时达高峰(3560%±174%,P<001);001~1 μmol/L孟鲁司特和HAMI 3379均能轻度抑制缺糖缺氧及恢复诱导的活性氧产生(均P<005)。缺糖缺氧及恢复诱导神经元线粒体膜电位下降,001~1 μmol/L孟鲁司特一定程度抑制该作用(P<005),而HAMI 3379无明显作用。外源性活性氧过氧化氢呈浓度和时间依赖性损伤神经元,01~1 μmol/L的孟鲁司特和0001~01 μmol/L的HAMI 3379能轻度减轻过氧化氢诱导的损伤(均P<005)。CysLT1R小分子干扰RNA和CysLT2R短发卡RNA对以上反应均无显著影响。结论:孟鲁司特和HAMI 3379在神经元缺血性损伤中均有轻度抗氧化作用,且此作用可能不依赖受体。

关键词 白三烯拮抗剂/药理学;神经元/药物作用创伤和损伤;乙酸盐类/治疗应用;喹啉类/治疗应用;氧化性应激    
Abstract

Objective: To investigate the antioxidative effects of two cysteinyl leukotriene receptors antagonists (CysLT1R and CysLT2R) montelukast and HAMI 3379 on ischemic injury of rat cortical neurons in vitro. Methods: Cultured rat cortical neurons were pretreated with CysLT1R antagonist montelukast and CysLT2R antagonist HAMI 3379, and then exposed to oxygenglucose deprivation/recovery (OGD/R) or H2O2. Reactive oxygen species (ROS), mitochondrial membrane potential (MMP) depolarization, neuronal viability and lactate dehydrogenase (LDH) release were determined. Meanwhile, RNA interference was used to inhibit the expression of CysLT1R and CysLT2R, and the effects were observed. Results: ROS production in neurons was significantly increased after 1 h OGD, which reached the peak at 30 min and lasted for 15 h after recovery. Montelukast and HAMI 3379 at 0011 μmol/L moderately decreased OGD/Rinduced ROS production (P<005). Montelukast mildly attenuated OGD/Rinduced MMP depolarization (P<005), but HAMI 3379 had no effect. H2O2 reduced neuronal viability and increased LDH release, namely inducing neuronal injury. Montelukast and HAMI 3379 at 011 μmol/L moderately attenuated H2O2induced neuronal injury (P<005). However, both CysLT1R siRNA and CysLT2R shRNA did not significantly affect the responses mentioned above. Conclusion: In ischemic neuronal injury, montelukast and HAMI 3379 exert a moderate antioxidative effect, and this effect may be receptorindependent.

Key wordsLeukotriene antagonists/pharmacology    Neurons/drug effects    Wounds and injuries    Acetates/therapeutic use    Quinolines/therapeutic use    Oxidative stress
收稿日期: 2013-12-01     
基金资助:

国家自然科学基金(81273491,81072618,81173041)

通讯作者: 魏尔清(1952-),男,博士,教授,博士生导师,从事神经药理学研究;Email: weieq2006 @zju.edu.cn   
Corresponding author: WEI Erqing, Email: weieq2006 @zju.edu.cn   
作者简介: 徐栋敏 (1989-),男,硕士研究生,从事神经药理学研究;Email∶xudongminxu@163.com
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引用本文:

徐栋敏,张霞燕,王晓蓉,等. 孟鲁司特和HAMI 3379对体外大鼠皮层神经元[J]. 浙江大学学报(医学版), 2014, 43(3): 257-264.
XU Dong-min, ZHANG Xia-yan, WANG Xiao-rong, CHEN Lu, ZHANG Li-hui, SHI Qiao-juan, FANG San-hua, LU Yun-bi, ZHANG Wei-ping, WEI Er-qing. Antioxidative effects of cysteinyl leukotriene receptor antagonists montelukast and HAMI 3379 on ischemic injury in rat cortical neurons in vitro. Journal of ZheJiang University(Medical Science), 2014, 43(3): 257-264.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.10089292.2014.05.001      或      http://www.zjujournals.com/xueshu/med/CN/Y2014/V43/I3/257

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