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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。
OBJECTIVE: Different fluid resuscitation was performed in the early stage of severe hemorrhagic shock in rats. The levels of Tregs, Th1 / Th2 and Tc1 / Tc2 in spleen tissues were compared. Of the different changes, preliminary study of its immune repair mechanism. Innovative points: (1) The spleen is an important immune organ of the body to detect changes in immune cells which are more sensitive and specific than peripheral blood; (2) the coordinated analysis of immune cell changes in the immune response is more Innovative and scientific, clinically standardized treatment programs provide a scientific and practical information. Methods: SD male rats were randomly divided into 5 groups. Control group and Sham group (sham operation) were compared only. The other three groups were given different liquid resuscitation after establishment of severe hemorrhagic shock rat model: isotonic saline NS group), hypertonic saline (HTS group) and hydroxyethyl starch (HES group). Thirty minutes later, the hemodynamics were monitored continuously for 120 minutes. The final cardiac puncture was taken and the spleen tissues were taken for further analysis of the contents of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells by Thymus fluorescence labeling flow cytometry, Th2 and Tc1 / Tc2 ratio. Conclusion: The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Tregs, Th1 / Th2 and Tc1 / Tc2 in the spleen of rats after liquid resuscitation were not significantly different between the control group, Sham group and HTS group Group and HES group. Compared with Sham group, Tc1 level in HTS group increased significantly, while Tc2 level increased in NS group, HES group and HTS group, and there was no difference in Tc2 level between the three groups. Thus, the effect of HTS fluid resuscitation on the immune system is greater than that of NS and HES for maintaining the balance of Treg cell content in the spleen, Th1 / Th2 and Tc1 / Tc2 balance. Taken together, HTS resuscitation provides potential immune repair in the early stages after hemorrhagic shock.