小鼠深Ⅱ度烫伤创面模型建立及创面缺氧情况检测
소서심Ⅱ도탕상창면모형건립급창면결양정황검측
Reproduction of a mouse model of deep partial-thickness scald and determination of hypoxia in the wound
  • 万方数据
    中华烧伤杂志 중화소상잡지
    16
    2013年 3期 277-280 ,共4页

    张月%白晓智%芦帅%李娜%王耘川%李小强%胡晓龙%胡大海

    장월%백효지%호수%리나%왕운천%리소강%호효룡%호대해

    烧伤%模型,动物%缺氧 燒傷%模型,動物%缺氧
    소상%모형,동물%결양
    Burns%Models,animal%Anoxia
    目的 建立稳定的小鼠深Ⅱ度烫伤创面模型并了解创面缺氧情况. 方法 (1)取80只体质量为(20.0±1.0)g的雄性BALB/c小鼠,将自制蒸汽烫伤仪喷头垂直置于小鼠背部表面直径为2 cm环形模具中心上方约2 cm处,用稳定的92℃蒸汽致伤2、4、6、8 s(每种致伤时间20只小鼠)制作烫伤模型.伤后即刻裸眼观察创面大体情况.于伤后0、12、24、48 h分别切取5只各致伤时间小鼠皮肤样本,HE染色行组织学观察,筛选深Ⅱ度创面致伤时间及取材时间.(2)另取5只BALB/c小鼠,按照前述方法与筛选致伤时间制成深Ⅱ度烫伤创面模型,伤后72 h用免疫组织化学染色法观察皮下组织缺氧情况.取20只BALB/c小鼠,按随机数字表法分为2组:正常对照组5只,不行处理；深Ⅱ度烫伤组15只,按照前述方法与筛选致伤时间制成深Ⅱ度烫伤创面模型.伤后72 h采用激光多普勒经皮氧分压仪检测创面中心、创面边缘和创面周边正常皮肤皮下氧含量,每个检测部位5只小鼠；正常对照组小鼠行相同检测.对数据行单因素方差分析. 结果 (1)伤后即刻,各种致伤时间小鼠创面苍白干净,无渗出.(2)组织学观察显示,烫伤深度随着致伤时间和取材时间的延长逐渐加深,至伤后24 h趋于稳定.烫伤4 s小鼠伤后24 h皮肤真皮深层受损,毛囊残留,呈现深Ⅱ度烫伤病理结果.(3)伤后72 h,小鼠深Ⅱ度烫伤创面可见哌莫硝唑高密度染色,创面边缘部位染色最明显.深Ⅱ度烫伤组小鼠创面中心、创面边缘、创面周边正常皮肤皮下氧分压相近,分别为(36.2±3.2)、(37.0±1.4)、(37.4±2.7)mm Hg(1 mm Hg=0.133 kPa,F=74.705,P＞0.05),均显著低于正常对照组的(53.1±2.4)mm Hg(F值分别为82.377、91.375、100.531,P值均小于0.05). 结论 采用体质量为(20.0±1.0)g的雄性BALB/c小鼠,经92℃蒸汽烫伤4 s,24 h后可建立相对稳定的深Ⅱ度烫伤创面模型.深Ⅱ度烫伤创面为缺氧环境,创面边缘部位缺氧最明显.
    목적 건립은정적소서심Ⅱ도탕상창면모형병료해창면결양정황. 방법 (1)취80지체질량위(20.0±1.0)g적웅성BALB/c소서,장자제증기탕상의분두수직치우소서배부표면직경위2 cm배형모구중심상방약2 cm처,용은정적92℃증기치상2、4、6、8 s(매충치상시간20지소서)제작탕상모형.상후즉각라안관찰창면대체정황.우상후0、12、24、48 h분별절취5지각치상시간소서피부양본,HE염색행조직학관찰,사선심Ⅱ도창면치상시간급취재시간.(2)령취5지BALB/c소서,안조전술방법여사선치상시간제성심Ⅱ도탕상창면모형,상후72 h용면역조직화학염색법관찰피하조직결양정황.취20지BALB/c소서,안수궤수자표법분위2조:정상대조조5지,불행처리；심Ⅱ도탕상조15지,안조전술방법여사선치상시간제성심Ⅱ도탕상창면모형.상후72 h채용격광다보륵경피양분압의검측창면중심、창면변연화창면주변정상피부피하양함량,매개검측부위5지소서；정상대조조소서행상동검측.대수거행단인소방차분석. 결과 (1)상후즉각,각충치상시간소서창면창백간정,무삼출.(2)조직학관찰현시,탕상심도수착치상시간화취재시간적연장축점가심,지상후24 h추우은정.탕상4 s소서상후24 h피부진피심층수손,모낭잔류,정현심Ⅱ도탕상병리결과.(3)상후72 h,소서심Ⅱ도탕상창면가견고막초서고밀도염색,창면변연부위염색최명현.심Ⅱ도탕상조소서창면중심、창면변연、창면주변정상피부피하양분압상근,분별위(36.2±3.2)、(37.0±1.4)、(37.4±2.7)mm Hg(1 mm Hg=0.133 kPa,F=74.705,P＞0.05),균현저저우정상대조조적(53.1±2.4)mm Hg(F치분별위82.377、91.375、100.531,P치균소우0.05). 결론 채용체질량위(20.0±1.0)g적웅성BALB/c소서,경92℃증기탕상4 s,24 h후가건립상대은정적심Ⅱ도탕상창면모형.심Ⅱ도탕상창면위결양배경,창면변연부위결양최명현.
    Objective To reproduce a stable mouse model of deep partial-thickness scald and to determine the hypoxia status in the wound.Methods (1) A homemade scald-producing apparatus with constant steam (92 ℃) emission was used to reproduce scald injury on the back (2 cm in diameter) in 80male BALB/c mice for different duration (2,4,6,and 8 s),with 20 mice for each scald duration.The nozzle was aligned perpendicularly to the back of mice,2 cm above the skin surface.The gross condition of wound was observed with naked eyes immediately after injury.Skin samples of 5 mice with different burn duration were harvested 0,12,24,and 48 h after scald for histopathological observation with hematoxylin and eosin staining,to screen the scalding time and time for biopsy of scalded skin to determine proper scalding time for the experiment.(2) Model of deep partial-thickness scald was reproduced with the desired scalding time as shown in the preliminary experiment in another 5 BALB/c mice.The hypoxia status in subcutaneous tissue was observed with immunohistochemical staining 72 h after scald.Another 20 BALB/c mice were divided into normal control group (n =5,without scald) and deep partial-thickness scald group (n =15,scalded for a suitable duration as determined in the preliminary experiment) according to the random number table.The subcutaneous oxygen content in wound center,the margin of the wound,and the normal skin adjacent to the wound was detected with laser Doppler transcutaneous oxygen tension 72 h after scald,with 5 mice in each region.Data were processed with one-way analysis of variance.Results (1) The wound of mice with different scald durations was pale,clean,and no exudate was observed right after injury.(2) The burn depth developed gradually along with the scalding time and sample harvesting time,and it became stable 24 h after scalding.A deep partial-thickness injury was observed in the dermis of mice scalded for 4 s and harvested 24 h after scald,and it was shown that the external hair sheath was still present,and it was determined to be a deep partial-thickness scald.(3) Dense staining of pimonidazole (hypoxia) was found in deep partial-thickness scald wound 72 h after scald,especially in the marginal zones of the wounds.The partial oxygen pressure in the wound center,wound margin,and normal skin around the wound was respectively (36.2±3.2),(37.0±1.4),(37.4 ±2.7) mm Hg (1 mm Hg=0.133 kPa),showing no statistically significant difference among them (F =74.705,P ＞ 0.05),but they were significantly lower than that of the control group [(53.1 ±2.4) mm Hg,with F values respectively 82.377,91.375,100.531,P values all below 0.05].Conclusions Deep partial-thickness scald model can be reproduced in (20.0 ± 1.0) g male BALB/c mice by scalding with 92 ℃ hot steam for 4 s,and the depth of wound becomes stable 24 h after scalding.Hypoxia can be found in the scalded wounds,especially in the marginal zones of the wounds.
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