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Immune recovery after fluid resuscitation in rats with severe hemorrhagic shock

严重失血性休克大鼠液体复苏后的免疫修复

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Abstract

Objective

To investigate the effects of resuscitation with normal saline (NS), hypertonic saline (HTS), and hydroxyethyl starch (HES) on regulatory T cells (Tregs), helper T 1 (Th1)/Th2 and cytotoxic T 1 (Tc1)/Tc2 profiles in the treatment of hemorrhagic shock.

Methods

Rats subjected to severe hemorrhagic shock were resuscitated for 30 min with NS (n=8), HTS (n=8), or HES (n=8); sham (n=8) and naive control (n=8) groups were used for comparison. Following fluid resuscitation, the whole shed blood was reinfused for 30 min, and the rats were observed with continuous hemodynamic monitoring for 120 min. CD4+CD25+Foxp3+ Treg proportions, Th1/Th2 and Tc1/Tc2 profiles in spleen were analyzed by three-color flow cytometry.

Results

The proportion of CD4+CD25+Foxp3+ Tregs and ratios of Th1/Th2 and Tc1/Tc2 did not differ among control, sham, and HTS groups, but were significantly lower in NS and HES groups (both P<0.05 vs. sham); NS and HES levels were similar. The level of Tc1 was significantly increased in HTS (P<0.05 vs. sham), and levels of Tc2 were increased in NS, HES, and HTS groups compared to sham (all P<0.05), but did not differ from each other.

Conclusions

HTS resuscitation has a greater impact on immune system recovery than NS or HES by preserving the proportion of Tregs and maintaining the balance between Th1/Th2 and Tc1/Tc2 cells in the spleen. Thus, HTS resuscitation provides potential immunomodulatory activity in the early stage after hemorrhagic shock.

摘 要

目 的

严重失血性休克大鼠模型早期阶段使用不同的液 体复苏, 比较脾脏组织中调节性T 细胞(Tregs)、 辅助性T 细胞1(Th1)/Th2 以及细胞毒性T 细 胞1(Tc1)/Tc2 的不同变化, 初步探讨其免疫修 复机制。

创新点

(1)脾脏为机体重要免疫器官, 检测其中的免 疫细胞变化, 比外周血更具敏感性和特异性; (2)将免疫反应中多环节的免疫细胞变化进行 协同分析, 结果更具创新性和科学性, 为临床上 形成规范的救治方案提供了科学的实践资料。

方 法

将SD雄性大鼠随机分成5 组, 其中对照组和Sham 组(假手术)仅作为比较, 其余三组在建立严重 失血性休克大鼠模型后, 采用不同的液体复苏: 等渗盐水(NS 组)、高渗盐水(HTS 组)和羟 乙基淀粉(HES 组)。然后再灌注30 分钟, 并 持续监测血液动力学120 分钟, 最后心脏穿刺, 取脾脏组织, 通过三色荧光标记流式细胞术进一步分析CD4+CD25+Foxp3+ Treg 细胞含量, 以及 Th1/Th2 和Tc1/Tc2 的比值。

结 论

液体复苏后大鼠脾脏中CD4+CD25+Foxp3+ Tregs 细胞含量、Th1/Th2 和Tc1/Tc2 的比值在对照组、 Sham 组和HTS 组中无差异, 并都显著高于NS 组和HES 组。与Sham 组比较, HTS 组中Tc1 水 平明显升高, 而NS 组、HES 组和HTS 组中Tc2 水平均有升高, 且三组之间Tc2 水平无差别。因 此, 对于维持脾脏中Treg 细胞含量、Th1/Th2 和 Tc1/Tc2 平衡的作用上, HTS 液体复苏对免疫系 统的影响大于NS 和HES。综上所述, 在失血性 休克后的早期阶段HTS 复苏可提供潜在的免疫 修复作用。

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Authors and Affiliations

  1. Department of Emergency Medicine, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310003, China

    Feng Yao, Yuan-qiang Lu & Jiu-kun Jiang

  2. Cancer Institute, Zhejiang Tumor Hospital, Hangzhou, 310022, China

    Lin-hui Gu & Han-zhou Mou

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  1. Feng Yao
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  2. Yuan-qiang Lu
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Correspondence to Yuan-qiang Lu.

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Project supported by the National Natural Science Foundation of China (No. 81272075)

ORCID: Yuan-qiang LU, http://orcid.org/0000-0002-9057-4344

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Yao, F., Lu, Yq., Jiang, Jk. et al. Immune recovery after fluid resuscitation in rats with severe hemorrhagic shock. J. Zhejiang Univ. Sci. B 18, 402–409 (2017). https://doi.org/10.1631/jzus.B1600370

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  • DOI: https://doi.org/10.1631/jzus.B1600370

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