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中华损伤与修复杂志(电子版)

中华损伤与修复杂志(电子版) ›› 2019, Vol. 14 ›› Issue (02) : 118 -123. doi: 10.3877/cma.j.issn.1673-9450.2019.02.008

所属专题： 文献；

论著

Nrf2/HO-1在远隔缺血后处理减轻大鼠心肺复苏脑损伤中的作用
李娟1, 白卫平2, 刘永飞1, 韩瑞丽1, 孙绪德1, 高昌俊1,†()   
  1. 1. 710038　西安，空军军医大学唐都医院麻醉科
    2. 714000　渭南市骨科医院麻醉科
  • 收稿日期:2019-01-15 出版日期:2019-04-01
  • 通信作者: 高昌俊
  • 基金资助:
    国家自然科学基金(81571183)

Effect of Nrf2 / HO-1 in remote ischemic post-conditioning alleviating rat brain injury induced by cardiopulmonary resuscitation

Juan Li1, Weiping Bai2, Yongfei Liu1, Ruili Han1, Xude Sun1, Changjun Gao1,†()   

  1. 1. Department of Anesthesiology, Tangdu Hospital, Air Force Military Medical University, Xi’an 710038, China
    2. Department of Anesthesiology, Weinan Orthopaedic Hospital, Weinan 714000, China
  • Received:2019-01-15 Published:2019-04-01
  • Corresponding author: Changjun Gao
  • About author:
    Corresponding auther: Gao Changjun, Email:
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引用本文：

李娟, 白卫平, 刘永飞, 韩瑞丽, 孙绪德, 高昌俊. Nrf2/HO-1在远隔缺血后处理减轻大鼠心肺复苏脑损伤中的作用[J]. 中华损伤与修复杂志(电子版), 2019, 14(02): 118-123.

Juan Li, Weiping Bai, Yongfei Liu, Ruili Han, Xude Sun, Changjun Gao. Effect of Nrf2 / HO-1 in remote ischemic post-conditioning alleviating rat brain injury induced by cardiopulmonary resuscitation[J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2019, 14(02): 118-123.

目的

探讨核因子E2相关因子2(Nrf2)/血红素氧化酶-1 (HO-1)在远隔缺血后处理减轻大鼠心肺复苏脑损伤过程中的表达变化及意义。

方法

采用随机数字表法将45只成年雄性SD大鼠分为3组：假手术组(Sham组)、模型组(Control组)、远隔缺血后处理组(RIPost组)，每组15只。Control组大鼠建立窒息性心肺复苏脑损伤模型，RIPost组大鼠在自主循环恢复后行肢体远隔缺血后处理。在自主循环恢复后24 h分别采用苏木精-伊红染色、酶联免疫吸附试验检测血清神经元特异性烯醇化酶(NSE)，观察脑组织的损伤情况；Western Blotting、免疫组织化学染色、实时荧光定量PCR检测脑组织Nrf2、HO-1蛋白及其mRNA表达情况。

结果

与Sham组相比，Control组呈急性缺血性改变，海马区神经元有明显的损伤，血清NSE含量显著升高，差异有统计学意义(P<0.05)，而RIPost组病理学改变轻于Control组，血清NSE含量也较Control组明显降低，差异有统计学意义(P<0.05)；Control组、RIPost组的Nrf2核蛋白和HO-1蛋白表达均高于Sham组，且RIPost组HO-1蛋白表达显著高于Control组，差异均有统计学意义(P值均小于0.05)。实时荧光定量PCR显示Control组及RIPost组Nrf2 mRNA表达水平差异无统计学意义(P>0.05)；RIPost组HO-1 mRNA的表达较Control组升高更显著，差异有统计学意义(P<0.05)。

结论

远隔缺血后处理可减轻大鼠心肺复苏脑损伤，其作用机制可能与上调大鼠Nrf2 /HO-1通路有关。

关键词: 心肺复苏术, 脑损伤, 血红素加氧酶-1, 远隔缺血后处理, 转录因子核因子E2相关因子2
Objective

To investigate the expression alternation and the effect of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in rats with remote ischemic post-conditioning alleviating brain injury induced by cardiopulmonary resuscitation.

Methods

According to the random number table method, a total of 45 adult male SD rats were divided in to Sham-operation group (Sham group), asphyxial cardiac arrest resuscitation group (Control group) and remote ischemic post-conditioning group (RIPost group), with 15 cases in each group. The rats in Control group were induced by 8-min asphyxiation and resuscitated with a standardized method, The remote ischemic post-conditioning models were established in RIPost group on the basis of the Control group. At the 24th hour after return of spontaneous circulation, pathomorphological changes in brain tissues were detected by hematoxylin-eosin staining, and the contents of neuron specific enolase(NSE) in serum were assessed by enzyme linked immunosorbent assay, the protein and mRNA expression of Nrf2, HO-1 in brain tissues were detected by Western blotting, immunohistochemistry and real-time(RT) PCR.

Results

Compared with Sham group, acute ischemic changes were found in Control group, which hippocampal neurons were obviously damaged, and NSE concentration in serum was increased significantly (P<0.05); while the pathological changes and serum NSE concentration in RIPost group were milder than those in Control group (P<0.05); the protein expressions of nuclear Nrf2 and HO-1 in Control group and RIPost group were higher than those in Sham group, and the expressions in RIPost group were higher than those in Control group (with P values below 0.05). In addition, RT-PCR analysis showed that there were no significant differences in Nrf2 mRNA levels in Control group and RIPost group (P>0.05), but HO-1 mRNA expression in RIPost group was significantly higher than that in Control group(P<0.05).

Conclusion

Remote ischemia post-conditioning can relieve brain injury induced by cardiopulmonary resuscitation, and the neuroprotective mechanism of RIPost may be related with the Nrf2/HO-1 signaling pathway.

Key words: Cardiopulmonary resuscitation, Brain injuries, Heme oxygenase-1, Remote ischemic post-conditioning, Nuclear factor E2-related factor 2
图1 各组大鼠海马CA1区神经元形态(苏木精-伊红染色，×400)。A示Sham组脑细胞胞膜完整，核仁清晰，苏木精染色均匀，无明显病理学改变；B示Control组脑细胞呈急性缺血性改变，细胞核固缩或溶解，细胞浆出现"嗜伊红"染色或正常细胞结构消失甚至出现空泡；C示RIPost组脑组织病理学改变较Control组明显减轻；Sham组：假手术组；Control组：模型组；RIPost组：远隔缺血后处理组
表1 各组大鼠自主循环恢复后24 h海马神经元存活率、血清NSE含量和Nrf2 mRNA、HO-1蛋白及mRNA的相对表达量比较(n＝5, ±s)
组别 神经元存活率(%) NSE含量((μg/L) Nrf2 mRNA HO-1 HO-1 mRNA
Sham组 0.90±0.03 36.91±4.40 2.21±1.26 0.33±0.36 0.88±0.27
Control组 0.49±0.08a 196.20±66.74a 1.73±0.91 0.46±0.35 1.13±0.25
RIPost组 0.67±0.07b 75.55±37.09b 1.15±0.41 1.00±0.32b 2.60±1.90b
图2 各组大鼠损伤脑组织中Nrf2蛋白的表达
图3 各组大鼠损伤脑组织中Nrf2表达(免疫组织化学染色，×400)。A示Sham组脑组织Nrf2表达(±)；B示Control组脑组织Nrf2表达较Sham组增强(＋)；C示RIPost组较Control组Nrf2表达增加更明显，且多位于细胞核中(＋＋)；Nrf2为核因子E2相关因子2；Sham组：假手术组，Control组：模型组，RIPost组：远隔缺血后处理组
图4 Western Blotting检测各组大鼠损伤脑组织中HO-1蛋白的表达。HO-1为血红素氧合酶-1；Sham组：假手术组，Control组：模型组，RIPost组：远隔缺血后处理组
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