全血血栓与白色血栓诱导的肺栓塞小鼠模型的比较研究

Comparative study on pulmonary embolism in mice induced by whole blood thrombus or white thrombus

目的对采用全血血栓及白色血栓诱导的肺栓塞小鼠模型进行评估,为选择简便、经济、成功率高、接近真实病理状态的小鼠急性肺栓塞模型提供依据。方法健康雄性清洁级C57BL/6J小鼠122只,随机分为正常对照组(n=6)、假手术组(n=12)、全血血栓栓塞组(H组,n=30)及白色血栓栓塞组(W组,n=30)。小鼠麻醉后,分离暴露右侧颈静脉,栓塞组穿刺并注入自体全血血栓或白色血栓(每只动物40个栓子,每个栓子长1mm,直径1.05mm),造成急性血栓性肺栓塞;假手术组注入等体积生理盐水。测定各组体循环血压、心率、右心室收缩压(RVSP)、外周血TNF-α水平,并行病理组织学观察了解栓塞效果。结果小鼠肺栓塞建模成功率为100%。注栓后W组、H组及假手术组血压分别为55.5±4.8、58.2±4.2、67.2±2.6mmHg,与正常对照组(71.3±3.0mmHg)比较均明显降低(P<0.01),且W、H组明显低于假手术组(P<0.01),W组与H组比较未见明显差异(P>0.05);注栓后W组、H组及假手术组心率分别为372±38、360±39、370±38次/min,与正常对照组(440±25次/min)比较均明显降低(P<0.01),而前3组间比较差异无统计学意义。栓塞后12h W组、H组RVSP分别为29.6±1.7、30.9±1.9mmHg,显著高于假手术组及正常对照组(分别为19.3±1.9、19.6±1.6mmHg,P<0.01);至栓塞后48h W组RVSP(28.5±3.1mmHg)与栓塞后12h比较无统计学差异(P>0.05),与假手术组(21.1±1.2mmHg)及正常对照组(19.6±1.6mmHg)比较明显升高(P<0.01),H组RVSP(21.0±1.5mmHg)与栓塞后12h比较明显降低(P<0.01),与假手术组及正常对照组比较无统计学差异(P>0.05)。注栓后1、6、12、24h,假手术组、W组、H组外周血TNF-α水平差异无统计学意义(P>0.05)。Mallory磷钨酸苏木精(PTAH)及HE染色显示,栓子大多处于肺动脉段及亚段水平,栓塞切实可靠,且W组栓子48h自溶率(30%)明显低于H组(100%)。结论白色血栓诱导的小鼠急性肺栓塞模型病理状态稳定,优于全血血栓诱导的肺栓塞模型。

Objective To evaluate pulmonary embolism in mice induced by whole blood thrombus or white thrombus,in order to provide a basis for choosing murine model of acute pulmonary embolism,which is easy to reproduce,less expensive,closer to the real pathological state,and with a high success rate.Methods A total of 122 healthy and clean male C57BL/6J mice were divided randomly into normal control group（n=6）,sham-operation group（n=12）,whole blood thrombus group（H group,n=30）,and white thrombus group（W group,n=30）.After the mice were anesthetized,the right jugular vein was isolated and exposed.The prepared white or whole blood autologous thrombi were introduced into the jugular vain（40 thrombi for each animal;each 1mm in length,1.05mm in diameter） to reproduce acute pulmonary thromboembolism.For the sham-operation group physiological saline of the same volume was infused.The systemic blood pressure,heart rate,right ventricular systolic pressure（RVSP）,and TNF-α of the peripheral blood of each animal were then determined,and pathological examination was performed to check the effects of the embolism.Results The success ratio of the reproduction of the murine model of pulmonary embolism is 100%.After the introduction of the embolus,the blood pressure（BP） of W group,H group and sham-operation group was 55.5±4.8,58.2±4.2 and 67.2±2.6mmHg,respectively,which was significantly lower compared with that of the normal control group（71.3±3.0mmHg,P〈0.01）.There was no difference in BP between the W and H group（P〈0.05）.After injecting the emboli,the heart rate of the W,H,and sham-operation group was 372±38,360±39 and 370±38b/min,respectively,and they were significantly lower compared with the normal control group（440±25b/min,P〈0.01）;there was no difference between the first 3 groups.After 12h of embolization,the RVSP of W and H groups（29.6±1.7 and 30.9±1.9mmHg,respectively） was significantly higher than that of the sham-operation group and normal control group（19.3±1.9 and 19.6±1.6mmHg,respectively,P〈0.01）.After 48h of embolization,there was no difference in the RVSP（28.5±3.1mmHg） compared with that of 12h after embolization in the W group（P〈0.05）,However,there was a significant increase compared with the sham-operation group（21.1±1.2mmHg,P〈0.01） and normal control group（19.6±1.6mmHg,P〈0.01）.In H group,RVSP 48h after embolization（21.0±1.5mmHg） showed a significant lowering compared with that of 12h after embolization,but there was no difference（P〈0.05） compared with that of the sham-operation group and normal control group.The TNF-α content in peripheral blood showed no difference among the sham-operation group,W group,and H group at 1,6,12,and 24h after embolization（P〈0.05）.Mallory phosphotungstic acid haematoxylin and HE staining showed that most emboli were localized in the branches or sub-branches of pulmonary arteries,and embolization was authentic.Further,the autolysis rate of the emboli in W group（30%） was obviously lower than that in the H group（100%） after 48h.Conclusions The murine model of acute pulmonary embolism reproduced with white thrombus exhibits a more steady pathological state,and is better than that induced by whole blood thrombus.