慢性血栓栓塞性肺动脉高压实验动物模型的建立

Establishment of the experimental animal models of chronic thromboembolic pulmonary hypertension

目的:建立操作简便,成功率高,接近人体生理学的慢性血栓栓塞性肺动脉高压的动物模型。方法:①实验于2004-12/2005-03在北京大学第一医院动物实验中心完成。选用健康杂种犬12只。②给予健康杂种犬戊巴比妥静脉麻醉后,取血50mL,加入凝血酶静置制备成血栓。切开颈外静脉插入7F导管,经导管向颈外静脉内注射自体血栓适量,同时监测生理指标。③达到实验标准后,缝合静脉切口及皮肤切口,给予抗生素,动物饲养15,30,60d后,重复上述麻醉和注栓实验步骤。所有动物分别于第一次注栓前和注栓后第90天记录平均肺动脉压、肺毛细血管嵌压、心输出量、呼吸频率、潮气量、气体流速、胸内压。肺血管阻力=(平均肺动脉压-肺毛细血管嵌压)/心输出量。采用Amdur和Mead方法利用呼吸频率、潮气量、呼吸流速和胸内压计算肺气道阻力和肺动态顺应性。内皮素1水平测定用放射免疫非平衡法。④采用实验动物模型建立前后平均肺动脉压、肺血管阻力、肺气道阻力和肺动态顺应性,以及血浆内皮素1水平作为判定模型建立成功与否的标准。⑤显著性比较用配对t检验。结果:①动物模型建立成功率为83%(10/12)。②成活动物饲养第90天血浆内皮素1水平、肺动脉平均压、肺血管阻力、肺气道阻力明显高于注栓前犤(35.3±7.2)pg/L,(28±5)mmHg,(5.71±0.68)(mmHg·min)/L,(128.47±34.32)Pa/(L·s);(20.6±6.3)pg/L,(12±2)mmHg,(2.71±0.23)(mmHg·min)/L,(61.78±5.88)Pa/(L·s),P<0.05犦,肺动态顺应性明显低于注栓前犤(1.23±0.36),(1.68±0.20)mL/kPa,P<0.05犦。结论:用自体血栓连续注射法建立慢性血栓栓塞性肺动脉高压动物模型有较高的可行性。

AIM： To establish the experimental animal models with chronic thromboembolic pulmonary hypertension （CTEPH）, which has simple operation, high successful rate and similar to human physiology. METHODS： ① The experiment was done at the Center of Animal Experiment, First Hospital, Peking, University from December 2004 to March 2005. Twelve healthy hybrid dogs were selected. ② The 50 mL blood was gained after intravenous anesthesia with pentobarbital. The thrombus was made statically after adding the thrombase. The 7 F tube was inserted into external jugular vein after cutting the vein, and through the tube the suitable autologous thrombus was injected into the external jugular vein, meanwhile, to monitor the physiological index. ③ After reaching the experimental standard, the vein cut and skin cut were sutured, and gave the antibiotic. After raising 15, 30 and 60 days, the experimental procedures of anesthesia and injecting thrombus were repeated. The mean pulmonary artery pressure （MPAP）, pulmonary capillary wedge pressure （PCWP）, cardiac output （CO）, respiratory rate （RR）, tidal volume, gas flow-rate and chest pressure were recorded at the 90 days before and after the first injected with thrombus in all the animals. Pulmonary vascular resistance （mm Hg/L per minute） = （MPAP- PCWP）/CO. Lung resistance （RL） and dynamic compliance of the respiratory system （Cdyn） were calculated by Amdur and Mead method according to respiratory rate, tidal volume, respiration flow-rate and chest pressure. The level of endothelins 1 was detected by non-balance method of radio-immunity. ④ The MPAP, PVR, RL Cdyn and the level of plasm endothelins 1 were taken as the standard to establish the successful model or unsuccessful model before and after the establishment of experimental animal models. ⑤ The difference was compared by the pairing t-test. RESULTS： ① The successful rate of the establishment of animal models was 83%（10/12）. ②The level of plasm endothelins 1, MPAP, PVR and RL of the living animal at the 90 days after raising were significantly higher than those before injecting the thrombus [（35.3±7.2） pg/L,（ 28 ±5） mm Hg, （5.71±0.68） mm Hg/L per minute, （128.47±34.32） Pa/（L.s）; （20.6±6.3） pg/L, （12±2） mm Hg, （2.71±0.23） mm Hg/L per minute, （61.78±5.88） Pa/（L·s）, P 〈 0.05]. The Cdyn was significantly lower than that before injecting the thrombus [（1.23±0.36）, （1.68±0.20） mL/kPa, P 〈 0.05]. CONCLUSION： The establishment of the experimental animal models with CTEPH by successive injection of autologous thrombus has very high feasibility.