p62-Keap1-Nrf2信号通路在减轻NOD/SCID小鼠心肺复苏后心肌损伤中的作用

doi:doi:10.3969/j.issn.1002-1949.08.020

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摘要目的探讨p62-Keap1-Nrf2抗氧化信号通路在严重联合免疫缺陷小鼠心脏骤停（CA）心肺复苏(CPR)后心肌缺血-再灌注（I/R）损伤过程中的作用机制。方法应用免疫功能正常的雄性C57BL/6小鼠（80只）和严重联合免疫缺陷(NOD/SCID)小鼠（80只）复制高钾合并窒息心脏骤停心肺复苏模型，根据复苏后取材时间点不同将两种品系小鼠分别分为复苏前组和复苏后2 h组、复苏后12 h组、复苏后24 h组及复苏后48 h组。观察各组小鼠自主循环恢复率（ROSC）、自主呼吸恢复率及48 h存活率。Western blot检测不同时间点两种小鼠心肌细胞中p62-Keap1-Nrf2信号通路相关蛋白的表达水平。透射电镜观察复苏后不同时间点线粒体形态。结果 ①两种小鼠心脏骤停心肺复苏后ROSC率和自主呼吸恢复率差异无统计学意义（P>0.05），但NOD/SCID 小鼠较C57BL/6小鼠心脏骤停心肺复苏后48 h存活率明显降低（ P ＜0.05）。②与复苏前比较，C57BL/6小鼠心肌细胞p62和P-p62（Ser351）蛋白表达在复苏后2 h时降低（P＜0.01 ），复苏后12 h时升高（ P ＜0.01），复苏后24 h和48 h时差异均无统计学意义（P＞0.05），而在复苏后各时间点，NOD/SCID小鼠心肌细胞p62和P-p62（Ser351）蛋白表达呈逐渐增高趋势（ P ＜0.01）。并且，NOD/SCID小鼠在复苏后2、24、48 h时心肌细胞中p62和P-p62（Ser351）蛋白表达均较C57BL/6小鼠明显增高（ P ＜0.01）。与复苏前比较，C57BL/6小鼠和NOD/SCID小鼠在心脏骤停心肺复苏后各时间点心肌细胞Keap1蛋白和细胞核Nrf2蛋白表达均明显增高。此外，与C57BL/6小鼠比较，NOD/SCID小鼠在复苏后各时间点心肌细胞Keap1蛋白表达均明显增高，而在复苏后2、12、48 h时NOD/SCID小鼠心肌细胞细胞核Nrf2蛋白均呈明显高表达（ P ＜ 0.01 ）。③C57BL/6小鼠在心肺复苏后心肌I/R损伤2 h和12 h可见心肌组织中线粒体明显肿胀，大量自噬小体形成；而NOD/SCID小鼠在复苏后2 h和12 h也可见到线粒体明显肿胀，但自噬体形成却发生在复苏后24 h和复苏后48 h，并且自噬小体数目较少。结论 NOD/SCID小鼠由于存在线粒体自噬缺陷，其在心脏骤停心肺复苏后心肌I/R损伤早期并未及时启动线粒体自噬机制，机体主要通过p62-Keap1-Nrf2抗氧化信号通路来发挥保护作用。

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	谢吐秀
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	魏捷
	叶璐

关键词 ：免疫缺陷, 心肺复苏(CPR), 氧化应激, p62-Keap1-Nrf2, 自主循环恢复率（ROSC）, 心脏骤停后综合征（PCAS）

通讯作者: 吕菁君（1973-），女，博士后，主任医师，急诊科副主任，E-mail：Lvjingjun@whu.edu.cn。

作者简介: 谢吐秀（1990-），女，硕士研究生，E-mail：1649594326@qq.com。

引用本文:

谢吐秀，吕菁君，魏捷，叶璐. p62-Keap1-Nrf2信号通路在减轻NOD/SCID小鼠心肺复苏后心肌损伤中的作用[J]. 中国急救医学, 2019, 39(8): 802-808.
Xie Tu-xiu, Lyu Jing-jun, Wei Jie, Ye Lu. Effect of p62-Keap1-Nrf2 signaling pathway on reducing myocardial injury after cardiopulmonary resuscitation in NOD/SCID mice. Chinese Journal of Critical Care Medicine, 2019, 39(8): 802-808.

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http://www.cnemergency.org.cn/CN/doi:10.3969/j.issn.1002-1949.08.020 或 http://www.cnemergency.org.cn/CN/Y2019/V39/I8/802

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