NMDA受体NR2A、NR2B亚基在戊四氮诱导慢性癫痫大鼠中的作用

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摘要目的：探讨NMDA受体NR2A、NR2B亚基在戊四氮（PTZ）诱导慢性癫痫大鼠惊厥发作中的作用及机制。方法：48只雄性SD大鼠随机分为：Control组、PTZ组、PTZ+MK-801组、PTZ+NVP组、PTZ+Ifenprodil组；连续腹腔注射35 mg/kg PTZ 28 d，注射PTZ 0.5 h前，各组分别注射等量0.9%氯化钠溶液、0.1 mg/（kg·d）MK-801、5.0 mg/（kg·d） NVP、10 mg/（kg·d） Ifenprodil。按照Racine分级标准每天记录行为学评分。第29天处死动物，尼氏染色观察大鼠海马神经元丢失情况，Western blot法测定大鼠海马NR2A、NR2B亚基水平。结果：与PTZ组比，MK-801组和NVP组戊四氮诱导慢性癫痫大鼠的惊厥发作减轻，而Ifenprodil组无减轻。与Control组比，PTZ组海马神经元大量丢失；与PTZ组比，MK-801、NVP和Ifenprodil均能减少海马神经元丢失；其中，MK-801、NVP减轻神经元损伤效果显著。与对照组比，各组惊厥发作后海马NR2A、NR2B亚基水平均下降，PTZ组最明显；NR2A亚基水平，PTZ+NVP组高于PTZ组；NR2B亚基水平，各药物干预组均高于PTZ组。结论：非选择性抑制NMDA受体和选择性抑制NR2A亚基能够减轻慢性癫痫大鼠发作强度和海马神经元损伤，而选择性抑制NR2B亚基仅能减轻海马神经元损伤，这提示惊厥发作可能与NR2A亚基有关，而神经元损伤可能与NR2A、NR2B亚基均有关。

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	徐惠琴
	杜延茹
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	丁思琦
	郑荣远
	王新施
	王莉
	叶朦倩

关键词 ：癫痫, 戊四氮, NMDA受体, NR2A亚基, NR2B亚基, 大鼠

Abstract：Objective: To explore the role and mechanism of N-methyl-D-aspartic acid (NMDA) receptor NR2A and NR2B subunits in pentetrazol (PTZ) induced chronic epileptic rats. Methods: Forty-eight male SD rats were randomly divided into control group, PTZ group, PTZ+MK-801 group, PTZ+NVP group, and PTZ+Ifenprodil group. Rats were daily kindled by intraperitoneal injection of 35 mg/(kg·d) PTZ for 28 days. Half an hour before PTZ kindle, rats were given intraperitoneal injection of 3.5 mL/(kg·d) saline, 0.1 mg/(kg·d) MK-801, 5.0 mg/(kg·d) NVP, and 10 mg/(kg·d) Ifenprodil, respectively. Seizure scores according to Racine standard were recorded for 28 consecutive days. All rats were sacrificed on the 29th day. Nissl’s staining was used to evaluate neuron loss of hippocampal CA1, CA3, and DG areas. Western blot was used to detect the expression of NR2A, NR2B subunits. Results: Compared with the PTZ group, MK-801 and NVP could reduce the intensity of convulsion attack in pentetrazol induced chronic epileptic rats, while Ifenprodil produced no such effect. Compared with the control group, hippocampal neurons in the PTZ group lost significantly; while compared with the PTZ group, hippocampal neuron losing in MK-801, NVP, and Ifenprodil groups were light, of which, MK-801 and NVP were the lightest. Compared with the control group, the levels of both NR2A and NR2B subunits in hippocampal decreased after convulsion attack and the decline was most pronounced in the PTZ group. Level of NR2A subunit in PTZ+NVP group was higher than PTZ group, while levels of NR2B subunit in PTZ+MK-801, PTZ+NVP and PTZ+Ifenprodil groups were higher than PTZ group. Conclusion: Non-selective inhibition of NMDA receptor and selective inhibition of NR2A subunit could reduce the intensity of convulsion attack and improve hippocampal neuron losing in pentetrazol induced chronic epileptic rats, while selective inhibition of NR2B subunit could only improve hippocampal neuron losing. This study suggested that the convulsion attack could be related to NR2A subunit, and the damage of neurons could be related to both NR2A and NR2B subunit.

Key words： epilepsy pentetrazol NMDA receptors NR2A subunit NR2B subunit rats

收稿日期: 2018-06-10

基金资助:浙江省自然科学基金面上项目（LY15H090015）。

作者简介: 徐惠琴（1972-），女，浙江温州人，副教授，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2018/V48/I12/866

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