一种连续动态直接观察皮肤创口感染的小鼠模型  

A mice model for continuous, dynamic and direct observation of skin wound infection

作　　者：石有振[1] 王莹[1] 顾钏[1] 贾志刚[1] 傅秀军[1] 方勇[1] 俞为荣[1] 姚敏[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院烧伤整形科,201900

出　　处：《中华创伤杂志》2012年第9期854-858,共5页Chinese Journal of Trauma

基　　金：上海市科学技术委员会浦江人才计划基金资助项目（10PJ407000）;上海市卫生局基金资助项目（2009100）

摘　　要：目的拟建立一种稳定、可连续动态直接观察皮肤创口感染的动物模型。方法制作小鼠背部全层皮肤切割伤模型，即刻予以皮下缝合后按随机数字表法分为四组：A组小鼠接种50μl无菌PBS液（PBS对照组），B、C、D组（感染组）小鼠分别接种50斗l含1×10^6、1×10^8和1×10^10菌落形成单位（colonyformingunit，CFU）／ml可发光耐甲氧西林金黄色葡萄球菌（methicil—lin—resistantstaphylococcusaureus，MRSA）菌株悬液。观察接种后每组小鼠的饮食活动，并记录12，3，5，7，9，11，14d的平均体重及其死亡率；接种后各时相点采用以电荷耦合器件（charge—COU—pieddevice，CCD）为基础的成像系统记录小鼠创口的荧光素发光强度；接种后24h切取创口局部组织HE染色，观察各组小鼠创口炎症反应情况；记录每组小鼠创口愈合时间。结果（1）平均体重：A、B组小鼠无明显波动；C组逐渐减轻，第3天后逐渐增加，第14天增至接种前水平；D组逐渐减轻直至死亡。（2）死亡率：A、B组为零，C组第14天小鼠死亡率为10％，D组小鼠至接种后第3天死亡率为100％。（3）创口荧光素发光强度：A、B组自接种当天起均逐渐减弱，分别至第5，7天接近完全消失；C组自接种当天起至第14天无明显增强和减弱迹象；D组自接种当天起逐渐增强，且范围逐渐扩大直至死亡。（4）接种后24hHE染色：四组小鼠创口均有炎症细胞浸润，C、D组浸润更明显。（5）创口愈合时间：A、B组分别于接种后第5，7天愈合；C组至第14天仍未愈合，且切口长度及范围未见明显扩大与减小；D组创口范围自接种当天起逐渐扩大直至死亡。结论小鼠皮肤全层切割伤创口接种50μl含1×10^8CFU／ml可发光MRSA菌株悬液，可直接实时动态连续观测创口感染的发生与发展，该感染模型易于制作，稳定和重复性高。Objective To establish a stable an/maR model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were rep- licated. After immediate subcutaneous suture, the mice were randomly divided into four groups, ic, Group A was inoculated with 50 μl sterile PBS solution）, Groups B, C and D were inoculated with 50 μl suspension containing 1 × 106, 1 × 108 and 1 × 1010 colony forming unit （CFU）/ml bioluminescent methi- cillin-resistant staphylococcus aureus （MRSA） respectively. Then, the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points. The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device （CCD） based imaging system. Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group. Wound healing time of each group was also recorded. Results （ 1 ） Average weight： Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the pre- inoculation level at day 14; Group D lightened gradually until death. （2）Mortality： Groups A and B had no death; Group C had 10% deaths at day 14; Group D had 100% deaths. （3） Bioluminescent intensity of wounds： Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death. （4） HE staining at 24 hours after inocula- tion ： all the four groups showed inflammatory cell infiltration, especially in Groups C and D. （5） Wound healing time： wound healed at days 5 and 7 after inoculation in Gro

关 键 词：模型 动物 感染 耐甲氧西林金葡菌 

分 类 号：R63[医药卫生—外科学] R-332[医药卫生—临床医学]