文章摘要
异氟醚通过上调NMDA受体导致发育期大鼠海马神经元凋亡
Isoflurane induces apoptosis of developing hippocampal neurons by up-regulating NMDA receptor
  
DOI：10.12089/jca.2018.09.013
中文关键词: 异氟醚  NMDA受体  凋亡  新生  海马
英文关键词: Isoflurane  NMDA receptor  Apoptosis  Newborn  Hippocampus
基金项目:国家自然科学基金(81500982)
作者单位E-mail
赵以林 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科  
张雪 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科 xuezhang@hust.edu.cn 
李世勇 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科  
方茜 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科  
严婧 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科  
唐晓乐 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科  
罗爱林 430030,武汉市,华中科技大学同济医学院附属同济医院麻醉科  
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中文摘要:
      
目的 探讨异氟醚麻醉对新生大鼠海马神经元N-甲基-D-天冬氨酸（NMDA）受体亚基及凋亡的影响。

方法 新生1 d的SD大鼠36只,雌雄不拘,取海马组织进行原代培养。采用随机数字表法将培养海马神经元5 d的培养皿随机分为三组: 对照组(C组)、异氟醚组(I组)和 AP5+异氟醚组(A组)，每组12只。I组和A组培养的海马神经元放置在密闭箱中, 吸入1.5％异氟醚和纯氧, 吸入时间6 h, C组不做任何处理。于异氟醚麻醉结束后2、4、6、24 h(C组于相应时点), 提取海马神经元总mRNA和总蛋白, 采用RT-PCR法测定NR2A mRNA、NR2B mRNA和caspase-3 mRNA的表达量, 采用Westen blot法测定caspase-3蛋白含量。

结果 与C组比较, 异氟醚麻醉结束后2、4和6 h I组海马神经元NR2A mRNA和NR2B mRNA的表达量明显上调(P<0.05)。与C组和A组比较,I组异氟醚麻醉结束后2、4和6 h海马神经元caspase-3 mRNA的表达量、异氟醚麻醉后4和6 h海马caspase-3蛋白含量明显升高(P<0.05)。

结论 吸入1.5％异氟醚可能通过上调新生大鼠海马神经元NMDA受体导致发育神经元凋亡, 阻断NMDA受体可以预防异氟醚导致的发育期海马神经元凋亡。
英文摘要:
      
Ojective To investigate the effects of isoflurane anesthesia on NMDA receptor subunits and cleaved caspase-3 in hippocampal of neonatal rats.

Methods Thirty-six primary culture neurons were randomly divided into 3 groups (n = 12 each): control groups (group C), isoflurane group (group I) and AP5+isoflurane (group A). 1.5％ isoflurane with 100％ O2 was inhaled for 6 h in group I and group A. Group C received no treatment. The total mRNA and protein were extracted 2, 4, 6 and 24 h after isoflurane anesthesia. RT-PCR was employed to detect NR2A mRNA, NR2B mRNA and caspase-3 mRNA expression, and Western blot was used to detect cleaved caspase-3 level.

Results Compared with group C, the expressions of NR2A mRNA, NR2B mRNA were increased at 2, 4, 6 h after isoflurane anesthesia (P < 0.05). Compared with group C and group A, the level of caspase-3 mRNA 2, 4, 6 h after isoflurane anesthesia and cleaved caspase-3 4 h and 6 h after isoflurane anesthesia were up-regulated in group I (P < 0.05).

Conclusion Inhaled isoflurane may induce the apoptosis of developmental neurons by up-regulating the NMDA receptor of hippocampal neurons in neonatal rats, and blocking the NMDA receptor can prevent the apoptosis of hippocampal neurons induced by isoflurane.
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