Page 30 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第43卷第4期
·468 · Journal of Nanjing Medical University（Natural Sciences） 2023年4月

·基础研究·

洛伐他汀调节NMDA受体功能减缓NMDA兴奋性毒性损害

李 蓉 ，刘 露 ，朱夕陈 ，马 涛 2*
2
2
1
江南大学附属中心医院药剂科，神经内科，江苏 无锡 214002
1 2

［摘 要］ 目的：检测洛伐他汀（lovastatin，LOV）对N⁃甲基⁃D⁃天门冬氨酸（N⁃methyl⁃D⁃aspartate，NMDA）诱导的兴奋性毒性的
神经保护作用并探讨 LOV 调节 NMDA 受体功能在神经保护中的潜在机制。方法：培养的大鼠原代神经元细胞分未处理组、
LOV 组、NMDA 组、LOV+NMDA 组、谷氨酸（glutamate，Glu）组及Glu+APV（一种特异性NMDA 受体拮抗剂）组。免疫荧光染色
检测神经元形态，TUNEL分析检测神经元凋亡，免疫印迹测定蛋白水平，生物素化法检测细胞表面受体。结果：①与NMDA组
或Glu组少数幸存微管相关蛋白（microtubule⁃associated protein 2，MAP⁃2）阳性神经元相比，LOV+NMDA组和Glu+APV组MAP⁃
2阳性神经元的数量明显增多，神经元树突的数目和长度均明显增加（P < 0.001）；②与NMDA 组或Glu组TUNEL 阳性细胞显
著增多相比，LOV+NMDA组或Glu+APV组TUNEL阳性细胞显著减少（P < 0.001）；③与未处理组相比，NMDA组NMDA受体蛋
白（N⁃methyl⁃D⁃aspartate receptor，NR2B）表达显著减少（P < 0.001），而LOV预处理后增加NR2B蛋白表达（P < 0.05）；④生物素化
法检测细胞表面受体显示，NMDA 处理导致细胞表面大部分 NR2B 丢失（P < 0.001），LOV 预处理能显著减少 NMDA 诱导的
细胞表面 NR2B 丢失（P < 0.05）。进一步研究显示，NMDA 处理后，NR2B 在酪氨酸（tyrosine，Tyr）1472 位点的磷酸化下降
（P < 0.05），LOV预处理显著恢复Tyr1472位点的磷酸化（P < 0.05）。结论：LOV能减轻NMDA诱导的兴奋性毒性损害，这一作
用可能与其影响NMDA受体亚单位NR2B的细胞内吞和/或胞内降解，进而调节NR2B表面分布有关。
［关键词］ 洛伐他汀；NMDA受体；NR2B；兴奋性毒性；神经保护
［中图分类号］ R363.2 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）04⁃468⁃07
doi：10.7655/NYDXBNS20230404

Lovastatin protects neurons from the excitotoxicity of NMDA by regulating the function of
NMDA receptors
2
2
1
LI Rong ，LIU Lu ，ZHU Xichen ，MA Tao 2*
Department of Pharmacy，Department of Neurology，Jiangnan University Medical Center，Wuxi 214002，China
1 2
［Abstract］ Objective：To explore the neuroprotective effect of lovastatin（LOV）on N ⁃ methyl ⁃ D ⁃ aspartate（NMDA）induced
excitotoxicity and the potential mechanism of LOV in regulating the function of NMDA receptors in neuroprotection. Methods：The
primary cultured rat neurons were divided into the vehicle group，LOV group，NMDA group，LOV+NMDA group，glutamate（Glu）
group and Glu + APV（a specific NMDA receptor antagonist）group. Neuronal morphology was detected by immunofluorescence
staining，neuronal apoptosis was detected by TUNEL analysis，protein levels were detected by Western blotting，cell surface receptors
were detected by biotinylation. Results：①Compared with the few surviving microtubule⁃associated protein 2（MAP⁃2）positive neurons
in the NMDA group or the Glu group，the number of MAP⁃2 immunopositive neurons in the LOV+NMDA group and the Glu+APV
group was significantly increased，as well as the number and length of neuronal dendrites were significantly increased（P < 0.001）.
②Compared with the significantly increased TUNEL⁃positive cells in the NMDA group or the Glu group，the TUNEL⁃positive cells in
the LOV+NMDA group or the Glu+APV group were significantly decreased（P < 0.001）. ③Compared with the vehicle group，the
expression of N ⁃ methyl ⁃ D ⁃ aspartate receptor（NR2B）in the NMDA group was significantly decreased（P < 0.001），while LOV
pretreatment could increase the expression of NR2B when compared with the NMDA group（P < 0.05）. ④The cell surface receptor
biotinylation assay showed that NMDA treatment resulted in the loss of the most NR2B on the cell surface（P < 0.001），while LOV
pretreatment could significantly reduce the NMDA⁃induced loss of NR2B（P < 0.05）. Further studies showed that phosphorylation of

［基金项目］ 国家自然科学基金（81000561）
∗
通信作者（Corresponding author），E⁃mail：tmadoc@njmu.edu.cn