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中华损伤与修复杂志(电子版) ›› 2023, Vol. 18 ›› Issue (04) : 285 -292. doi: 10.3877/cma.j.issn.1673-9450.2023.04.003

论著

高温高湿环境暴露对重度放创复合伤小鼠损伤恢复的影响
汪国建, 谭雨龙, 龙爽, 吕晓凡, 赵娜, 冉新泽, 王军平, 王涛†()   
  1. 400038　重庆，陆军军医大学（第三军医大学）军事预防医学系全军复合伤研究所　创伤、烧伤与复合伤国家重点实验室
    400038　重庆，陆军军医大学（第三军医大学）热带医学教研室
  • 收稿日期:2023-04-27 出版日期:2023-08-01
  • 通信作者: 王涛
  • 基金资助:
    国家自然科学基金面上项目(82172219); 军队后勤科研项目(ALJ19J002、ZLJ22J020)

Effects of hot and humid environments exposure on recovery of mice subjected to severe radiation injury combined with skin-wound trauma

Guojian Wang, Yulong Tan, Shuang Long, Xiaofan Lv, Na Zhao, Xinze Ran, Junping Wang, Tao Wang†()   

  1. Institute of Combined Injury of PLA, State Key Laboratory of Trauma, Burn and Combined Injury
    Department of Tropical Medicine, College of Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing 400038, China
  • Received:2023-04-27 Published:2023-08-01
  • Corresponding author: Tao Wang
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引用本文：

汪国建, 谭雨龙, 龙爽, 吕晓凡, 赵娜, 冉新泽, 王军平, 王涛. 高温高湿环境暴露对重度放创复合伤小鼠损伤恢复的影响[J/OL]. 中华损伤与修复杂志(电子版), 2023, 18(04): 285-292.

Guojian Wang, Yulong Tan, Shuang Long, Xiaofan Lv, Na Zhao, Xinze Ran, Junping Wang, Tao Wang. Effects of hot and humid environments exposure on recovery of mice subjected to severe radiation injury combined with skin-wound trauma[J/OL]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2023, 18(04): 285-292.

目的

研究重度放创复合伤(R-W-CI)小鼠暴露于高温高湿环境(HHE)不同时间对小鼠存活率、造血损伤恢复、创面愈合及其病理学的影响。

方法

HHE模拟实验舱参数设定为温度(36±0.5) ℃，相对湿度(70±5)%，通过肛温测评小鼠HHE暴露的效果。采用60Co γ射线6.0 Gy全身照射，建立小鼠重度骨髓型急性放射病模型；照射后1.0 h内用刀片制成体表面积2.0%皮肤创伤模型。两种模型小鼠均于致伤前1.0 h放入模拟舱，转运与照射均由便携式高温高湿模拟舱维持HHE，致伤完毕后置于高温高湿模拟舱1.0 h和7.0 h后取出按常规条件饲养。急性放射损伤小鼠分为4组，即正常对照组(NC组)、放射损伤组(R组)、放射损伤HHE暴露1.0 h组(R+HHE 1.0 h组)、放射损伤HHE暴露7.0 h组(R+HHE 7.0 h组)。观察不同HHE暴露时间(1.0 h和7.0 h)对损伤后小鼠肛温、存活率、体重及外周血变化的影响。重度放创复合伤小鼠于伤后给予常规补液、创面包扎或左氧氟沙星(LEV)抗菌治疗，分为7组，即正常对照组(NC组)、放创复合伤组(R-W-CI组)、放创复合伤左氧氟沙星治疗组(R-W-CI+LEV组)、放创复合伤HHE暴露1.0 h组(R-W-CI+HHE 1.0 h组)、放创复合伤HHE暴露7.0 h组(R-W-CI+HHE 7.0 h组)、放创复合伤HHE暴露1.0 h左氧氟沙星治疗组(R-W-CI+HHE 1.0 h+LEV组)、放创复合伤HHE暴露7.0 h左氧氟沙星治疗组(R-W-CI+HHE 7.0 h+LEV组)。观察不同HHE暴露时间小鼠存活率、体重、外周血、创面愈合以及病理学指标变化。

结果

模拟实验舱设定HHE条件对小鼠的体温能够产生显著影响，达到HHE效果。HHE不同暴露时间对重度骨髓型急性放射病小鼠的体重恢复有不同影响，但对存活率无影响。在无左氧氟沙星抗菌治疗时，R-W-CI+HHE 1.0 h组小鼠存活率(40%)显著优于R-W-CI组(20%)(χ2＝5.496，P＝0.019)，但与R-W-CI+HHE 7.0组(30%)无显著差异。左氧氟沙星抗菌治疗能显著提高R-W-CI+LEV组与R-W-CI+HHE 1.0+LEV组小鼠存活率，但对R-W-CI+HHE 7.0+LEV组无明显影响。致伤后28 d，R-W-CI+HHE1.0 h+LEV组白细胞恢复显著优于R-W-CI+LEV组(F＝6.549，P＝0.021)和R-W-CI+HHE 7.0 h+LEV组(F＝11.901，P＝0.05)。R-W-CI+HHE 7.0 h+LEV组小鼠创面愈合显著延迟，而R-W-CI+HHE 1.0 h+LEV组小鼠创面愈合显著加快。

结论

HHE短时间(1.0 h)暴露有利于重度放创复合伤恢复，HHE长时间(7.0 h)暴露与之相反；抗生素治疗是有助于改善放创复合伤预后的基础支持疗法。

关键词: 高温高湿环境, 放创复合伤, 造血, 创面愈合
Objective

To investigate the effects of hot and humid environments (HHE) exposure with different time on the recovery of mice suffered from radiation injury combined with skin-wound trauma (R-W-CI) by evaluating survival rate, hematopoietic injury recovery and cutaneous wound healing.

Methods

The parameters of the HHE simulation experiment module were set as 36 ℃±0.5 ℃ in temperature and 70%±5% in relative humidity, which were confirmed by detecting rectal temperature after mice were put in the module. The animal model of severe hematopoietic acute radiation sickness was established by mice with 6.0 Gy 60Co γ-rays whole body irradiation. The animal model of severe R-W-CI was established with mice exposed to 60Co γ-rays (6.0 Gy) followed by a 2.0% total-body-surface-area wounds within 1 h after irradiation. Mice from both models were set into HHE for 1 h before irradiation and transferred to one portable HHE simulation chamber during transportation and irradiation. The injured mice were continued to be put into HHE module for another 1.0 h or 7.0 h followed by maintaining conventional ambient temperature and humidity conditions. For hematopoietic acute radiation sickness model, the mice were divided into four groups: normal control (NC group), radiation injury (R group), radiation injury exposed to HHE for 1.0 h(R+ HHE1.0 h group), radiation injury exposed to HHE for 7.0 h(R+ HHE7.0 h group). Indexes including rectal temperature, survival rate, body weight and complete blood count were measured to estimate the effects of different HHE exposure time on the recovery of radiation injury. For R-W-CI model, routine fluid supplement and wound bandaging or levofloxacin (LEV) were implemented as routine supportive treatments. The mice were divided into seven groups: normal control(NC group), combined radiation-trauma injury (R-W-CI group), combined radiation-trauma injury treated with levofloxacin (R-W-CI+ LEV group), combined radiation-trauma injury exposed to HHE for 1.0 h(R-W-CI+ HHE 1.0 h group), combined radiation-trauma injury exposed to HHE for 7.0 h(R-W-CI+ HHE 7.0 h group), combined radiation-trauma injury exposed to HHE for 1.0 h and treated with levofloxacin (R-W-CI+ HHE 1.0 h+ LEV group), combined radiation-trauma injury exposed to HHE for 7.0 h and treated with levofloxacin (R-W-CI+ HHE 7.0 h+ LEV group). The HHE exposure effects on mice suffered from R-W-CI were evaluated by detecting survival rate, body weight, complete blood count and wound healing. In addition, the effects of antibacterial therapy with LEV in R-W-CI models were also evaluated.

Results

The simulation experiment module conducted the HHE with well effects, confirmed by significant increased rectal temperature of mice after put into the module. In model of severe hematopoietic acute radiation sickness, HHE exposure for 1.0 h and 7.0 h impacted body weight recovery but had no effect on survival rate. For R-W-CI model, in the absence of LEV treatment, survival rate in R-W-CI+ HHE 1.0 h group was significantly higher than that in R-W-CI group (χ2=5.496, P=0.019). LEV treatment could significantly increase the survival rate in both R-W-CI+ LEV group and R-W-CI+ HHE 1.0 h+ LEV group, but not in R-W-CI+ HHE 7.0 h+ LEV group. 28 days after injury, the recovery of white blood cells in R-W-CI+ HHE 1.0 h+ LEV group was significantly better than that in R-W-CI+ LEV group (F=6.549, P=0.021) and R-W-CI+ HHE 7.0 h+ LEV group (F=11.901, P=0.05). The wound healing in R-W-CI+ HHE 7.0 h+ LEV group was significantly delayed, while the wound healing in R-W-CI+ HHE 1.0 h+ LEV group was significantly accelerated.

Conclusion

Long-term (7.0 h) exposure to HHE after R-W-CI was detrimental, while short-term (1.0 h) exposure to HHE was beneficial to recovery of severe R-W-CI. The antibiotic therapy is important supportive treatment for R-W-CI exposed to HHE.

Key words: Hot and humid environments, Combined radiation-trauma injury, Hematopoiesis, Wound healing
图1 模拟实验舱内HHE暴露对正常小鼠和6.0 Gy全身照射小鼠体温变化的影响(**P<0.001)
图2 HHE暴露对6.0 Gy全身照射小鼠体重恢复的影响
表1 放创复合伤小鼠致伤后HHE不同暴露时间及左氧氟沙星治疗对小鼠存活率的影响(Generalized Wilcoxon Test)
组别   LEV(－)
  R-W-CI R-W-CI+HHE 1.0 h R-W-CI+HHE 7.0 h
  χ2值 P值 χ2值 P值 χ2值 P值
LEV(－) R-W-CI     5.495 0.019 0.120 0.729
  R-W-CI+HHE 1.0 h 5.495 0.019     3.007 0.083
  R-W-CI+HHE 7.0 h 0.120 0.729 3.007 0.083    
LEV(+) R-W-CI 11.33 0.001 11.33 0.001 8.633 0.003
  R-W-CI+HHE 1.0 h 11.33 0.001 6.374 0.012 4.499 0.034
  R-W-CI+HHE 7.0 h 0.055 0.814 0.641 0.423 0.014 0.907
组别   LEV(+)
  R-W-CI R-W-CI+HHE 1.0 h R-W-CI+HHE 7.0 h
  χ2值 P值 χ2值 P值 χ2值 P值
LEV(－) R-W-CI 11.33 0.001 11.33 0.001 0.055 0.814
  R-W-CI+HHE 1.0 h 11.33 0.001 6.374 0.012 0.641 0.423
  R-W-CI+HHE 7.0 h 8.633 0.003 8.633 0.003 0.014 0.907
LEV(+) R-W-CI     0.005 0.942 4.499 0.034
  R-W-CI+HHE 1.0 h 0.005 0.942     8.633 0.003
  R-W-CI+HHE 7.0 h 4.499 0.034 8.633 0.003    
图3 放创复合伤小鼠致伤后HHE不同暴露时间及左氧氟沙星治疗对小鼠存活率的影响
表2 放创复合伤小鼠致伤后HHE不同暴露时间及左氧氟沙星治疗对小鼠体重变化的影响(P值)
组别   NC LEV(－) LEV(+)
  R-W-CI R-W-CI+HHE 1.0 h R-W-CI+HHE 7.0 h R-W-CI R-W-CI+HHE 1.0 h R-W-CI+HHE 7.0 h
  NC   0.000 0.000 0.000 0.000 0.000 0.000
LEV(－) R-W-CI 0.000   0.437 0.834 0.055 0.041 0.420
  R-W-CI+HHE1.0 h 0.000 0.437   0.261 0.156 0.113 0.997
  R-W-CI+HHE7.0 h 0.000 0.834 0.261   0.013 0.009 0.239
LEV(+) R-W-CI 0.000 0.055 0.156 0.013   0.826 0.128
  R-W-CI+HHE1.0 h 0.000 0.041 0.113 0.009 0.826   0.090
  R-W-CI+HHE7.0 h 0.000 0.420 0.997 0.239 0.128 0.090  
图4 放创复合伤小鼠致伤后HHE不同暴露时间及左氧氟沙星治疗对小鼠体重变化的影响
表3 放创复合伤致伤后28 d小鼠外周血象的变化(±s)
组别 只数 白细胞(×109/L) 红细胞(×1012/L) 血小板(×109/L)
R-W-CI+LEV 9 14.43±2.94 11.01±0.91b 1805.33±380.95
R-W-CI+HHE 1.0 h+LEV 9 18.03±3.05ab 11.27±0.99b 1878.67±218.25
R-W-CI+HHE 7.0 h+LEV 4 12.36±1.60 9.81±0.63 1748.00±18.33
表4 放创复合伤致伤后28 d小鼠外周血白细胞分类计数的变化(±s)
组别 只数 淋巴细胞(×109/L) 中性粒细胞(×109/L) 单核细胞(×109/L)
R-W-CI+LEV 9 7.29±3.03 4.40±1.02 2.10±0.89
R-W-CI+HHE 1.0 h+LEV 9 9.71±1.56a 5.60±2.48 1.99±0.88
R-W-CI+HHE 7.0 h+LEV 4 6.84±1.51 2.00±0.54 1.98±1.10
图5 HHE不同暴露时间对放创复合伤创面愈合的影响。A示代表性图片；B示创面面积量化分析曲线；F＝885.441，**P<0.01
图6 HHE不同暴露时间对放创复合伤创面愈合后瘢痕组织影响的病理观察(HE)。A示各组代表性图片；B示瘢痕宽度量化比较；C示瘢痕面积量化比较；F＝9.913，*P＝0.012
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