CAJ | 학술논문

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50μl含1×106、1×108和1×1010菌落形成单位(colony forming unit,CFU)/ml可发光耐甲氧西林金黄色葡萄球菌(methicillin- resistant staphylococcus aureus,MRSA)菌株悬液.观察接种后每组小鼠的饮食活动,并记录12,3,5,7,9,11,14 d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 - couplddevice,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24 h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24 h HE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1×108 CFU/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.
목적 의건립일충은정、가련속동태직접관찰피부창구감염적동물모형. 방법 제작소서배부전층피부절할상모형,즉각여이피하봉합후안수궤수자표법분위사조:A조소서접충50μl무균PBS액(PBS대조조),B、C、D조(감염조)소서분별접충50μl함1×106、1×108화1×1010균락형성단위(colony forming unit,CFU)/ml가발광내갑양서림금황색포도구균(methicillin- resistant staphylococcus aureus,MRSA)균주현액.관찰접충후매조소서적음식활동,병기록12,3,5,7,9,11,14 d적평균체중급기사망솔；접충후각시상점채용이전하우합기건(charge - couplddevice,CCD)위기출적성상계통기록소서창구적형광소발광강도；접충후24 h절취창구국부조직HE염색,관찰각조소서창구염증반응정황；기록매조소서창구유합시간. 결과 (1)평균체중:A、B조소서무명현파동；C조축점감경,제3천후축점증가,제14천증지접충전수평；D조축점감경직지사망.(2)사망솔:A、B조위령,C조제14천소서사망솔위10％,D조소서지접충후제3천사망솔위100％.(3)창구형광소발광강도:A、B조자접충당천기균축점감약,분별지제5,7천접근완전소실；C조자접충당천기지제14천무명현증강화감약적상；D조자접충당천기축점증강,차범위축점확대직지사망.(4)접충후24 h HE염색:사조소서창구균유염증세포침윤,C、D조침윤경명현.(5)창구유합시간:A、B조분별우접충후제5,7천유합；C조지제14천잉미유합,차절구장도급범위미견명현확대여감소；D조창구범위자접충당천기축점확대직지사망. 결론 소서피부전층절할상창구접충50μl함1×108 CFU/ml가발광MRSA균주현액,가직접실시동태련속관측창구감염적발생여발전,해감염모형역우제작,은정화중복성고.
Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,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 methicillin-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 preinoculation 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 inoculation: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 Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.