[1]石　磊　周银芳　方　圆.褪黑激素对体外培养神经元七氟醚损害的保护作用[J].中国临床神经外科杂志,2021,26(05):354-359.[doi:10.13798/j.issn.1009-153X.2021.05.012]
　SHI Lei,ZHOU Yin-fang,FANG Yuan..Protective effect of melatonin on sevoflurane injury to cultured neonatal rat cortical neurons[J].,2021,26(05):354-359.[doi:10.13798/j.issn.1009-153X.2021.05.012]
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褪黑激素对体外培养神经元七氟醚损害的保护作用()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
26
期数:
2021年05期
页码:
354-359
栏目:
实验研究
出版日期:
2021-05-25

文章信息/Info

Title:
Protective effect of melatonin on sevoflurane injury to cultured neonatal rat cortical neurons
文章编号:
1009-153X（2021）05-0354-06
作者:
石　磊　周银芳　方　圆
725000 陕西，安康市中心医院麻醉科（石　磊、方　圆）；725000 陕西，安康市人民医院检验科（周银芳）
Author(s):
SHI Lei1;  ZHOU Yin-fang2;  FANG Yuan1.
1. Department of Anesthesiology, Ankang Central Hospital, Ankang 725000, China; 2. Department of Laboratory, Ankang People’s Hospital, Ankang 725000, China
关键词:
皮质神经元; 褪黑激素; miR-130a; ROCK2; 七氟醚; 大鼠乳鼠
Keywords:
Neonatal rat cortical neuron;  Melatonin;  miR-130a;  Sevoflurane
分类号:
R 36; R 614
DOI:
10.13798/j.issn.1009-153X.2021.05.012
文献标志码:
A
摘要:
目的 探讨褪黑激素（MLT）对体外培养神经元七氟醚（SEV）损伤的保护作用及其机制。方法 取SD大鼠乳鼠全脑皮层组织，分离皮质神经元进行体外培养；MLT预处理24 h后，4% SEV作用神经元；采用CCK-8法检测神经元存活率，流式细胞术检测神经元凋亡率，qPCR检测神经元miR-130a-3p和ROCK2 mRNA水平；免疫印迹法检测凋亡相关蛋白表达；应用在线预测软件TargetScan分析预测miR-130a-3p与ROCK2靶向关系并验证。结果 SEV明显降低神经元存活率以及miR-130a-3p和ROCK2 mRNA水平（P<0.05），明显增加神经元凋亡率及凋亡相关蛋白cle-caspase3和cle-caspase9表达水平（P<0.05）；MLT明显抑制SEV的作用（P<0.05）。软件TargetScan分析显示，miR-130a-3p与ROCK2的3’非编码区第846~852碱基处存在结合位点，PCR和免疫印迹法检测显示，miR-130a-3p与ROCK2存在靶向关系。ROCK2过表达明显逆转MLT预处理的效果（P<0.05）。结论 MLT预处理明显改善SEV对体外培养的大鼠乳鼠皮质神经元的损害，机制可能是上调miR-130a，进而靶向抑制ROCK2表达。
Abstract:
Objective To explore the protective effect of melatonin (MLT) on the sevoflurane (SEV) injury to cultured neonatal rat cortical neurons. Methods The neonatal rat cortical neurons were isolated from the whole cerebral cortex tissues of SD neonatal rats and cultured in vitro. After pretreatment with MLT for 24 h, the cortical neurons were treated with 4% SEV. The survival and apoptosis rates of neurons were detected using CCK-8 method and flow cytometry, respectively. The levels of miR-130a-3p and ROCK2 mRNA were detected by qPCR. The expression levels of apoptosis-related proteins were detected by Western blot. The targeted relationship between miR-130a-3p and ROCK2 was predicted using TargetScan software and verified. Results SEV significantly decreased the neuron survival rate and the levels of miR-130a-3p and ROCK2 mRNA (P<0.05), and significantly increased the neuron apoptosis rate and expression levels of apoptosis-related proteins including cle-caspase3 and cle-caspase9 (P<0.05). MLT significantly inhibited the effect of SEV on the cultured neurons (P<0.05). Software TargetScan analysis showed that miR-130a-3p and ROCK2 had a binding site at base 846~852 of the 3’non-coding region. PCR and Western blotting results showed that miR-130a-3p and ROCK2 had a targeting relationship. ROCK2 overexpression significantly reversed the effect of MLT pretreatment on the injured neurons induecd by SEV (P<0.05). Conclusions MLT pretreatment significantly improve the SEV injury to the cultured neonatal rat cortical neurons, which may be possibly through up-regulation miR-130a, and then targeted inhibition of ROCK2 expression.

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备注/Memo

备注/Memo:
基金项目：陕西省科技厅社会发展科技攻关项目（2016SF-246）
通讯作者：方　圆，E-mail：fangyuan110@163.com
更新日期/Last Update: 2021-05-25