摘要
目的探讨侵袭性肺曲霉病(IPA)小鼠的凝血功能异常及低分子肝素的治疗作用。方法采用环磷酰胺免疫抑制、烟曲霉孢子悬液滴鼻法构建中性粒细胞减少小鼠IPA模型。(1)凝血功能测定:72只小鼠随机分为正常免疫非接种组(A组)、正常免疫接种组(B组)、免疫抑制非接种组(C组)、模型组(D组),每组18只。烟曲霉孢子接种量为1.5×105个孢子/小鼠,接种后每24 h每组随机取6只小鼠进行出血时间、凝血时间、血小板计数及抗凝血酶Ⅲ(AT-Ⅲ)活性测定,共3次。(2)低分子肝素对IPA小鼠生存时间的影响:118只IPA模型小鼠随机分为模型对照组(E组,n=29)、低分子肝素治疗组(F组,n=30,1 000 IU/kg,背部皮下注射,qd×7 d)、两性霉素B治疗组(G组,n=29,1 mg/kg,腹腔注射,qd×7 d)、低分子肝素加两性霉素B治疗组(H组,n=30)。治疗均从接种后24 h开始。烟曲霉孢子接种量为6×105个孢子/小鼠。接种孢子后每天观察生存情况1次,至21 d。结果D组接种烟曲霉孢子后24 h即出现血浆AT-Ⅲ活性降低;48 h后出、凝血时间缩短,血浆AT-Ⅲ活性较24 h时降低;72 h后出现明显的血小板减少,出血时间进一步缩短,凝血时间较48 h延长,但仍低于正常,血浆AT-Ⅲ活性进一步降低;上述各指标变化D组与同时间点其他各组比较均有显著性差异(P<0.01),在各时间点A组、B组及C组之间比较均无显著性差异(P均>0.05)。生存分析结果表明,单独应用低分子肝素未提高IPA小鼠21 d存活率,亦未延长平均生存时间(F组与E组比较,P>0.05)。联合应用低分子肝素与两性霉素B疗效优于单独应用两性霉素B(G组与E组及F组比较,P均<0.05;H组与E组及F组比较,P均<0.01;H组与G组比较,P<0.05)。结论小鼠发生IPA后可出现凝血功能紊乱,血浆AT-Ⅲ活性可以作为IPA出现凝血功能紊乱的早期监测指标。与单独应用两性霉素B比较,低分子肝素与两性霉素B联用能够延长IPA小鼠的平均生存时�
Objective To investigate the blood clotting dysfunction of invasive pulmonary aspergillosis（IPA） and the therapeutic effect of low molecular heparin in a mouse model. Methods The neutropenic IPA mouse model was constructed by being given cyclophosphamide to depress immunologic function, and then intranasally challenged with Aspergillus fumigatus conidia. （ 1 ） Blood clotting function were assessed by bleeding time, clotting time, platelet count and antithrombase-Ⅲ （AT-Ⅲ） activity. Seventy-two mice were randomly assigned into 4 groups. Group A received only normal saline. Group B received normal saline to substitute the cyclophosphamide, and the rest equal to group D. Group C received normal saline to substitute the AspergiUus fumigatus conidia suspension, and the rest equal to group D. Group D （ model group） received cyclophosphamide （ intraperitoneally, 150 mg/kg, d4, d1 ） and Aspergillus fumigatus conidia suspension （ intranasally, 40 μL/mouse, 1.5 × 10^5/mL, dO ）. Six mice were randomly sacrificed in each group for analysis of blood clotting function per 24 h after inoculation for 3 times. （2） Therapeutic effect of low molecular heparin was determined by survival time of IPA mice. One hundred and eighteen mice were randomly assigned into 4 groups after challenged with 6 × 10^5 conidia/ mouse and received one of the following regimens daily from d1 to d7 after challenge,vehicle（ group E ,n = 29）, low molecular heparin （ group F, n = 30, subcutaneous injection, 1000 IU/kg, qd × 7 d）, amphotericin B （ group G, n = 29, intraperitoneal, 1 mg/kg, qd × 7 d）, low molecular heparin plus amphotericin B （ group H, n = 30）. Mice survivals were recorded once daily to d21 after innoculation. Results （ 1 ） AT-Ⅲ activity of group D decreased significantly 24 h after innoculation. Bleeding time and clotting time decreased significantly and AT-Ⅲ activity decreased sequentially 48 h after innoculation. The platelet decreased significantly 72 h after innoculation,and bl
出处
《中国呼吸与危重监护杂志》
CAS
2008年第6期440-444,共5页
Chinese Journal of Respiratory and Critical Care Medicine
基金
上海市卫生局科技发展基金资助项目(编号:044019)
关键词
侵袭性肺曲霉病
凝血功能
低分子肝素
弥散性血管内凝血
Invasive pulmonary aspergillosis
Blood clotting function
Low molecular heparin
Disseminated intravascular coagulation
