离体血管环张力系统标准化测定大鼠肺动静脉张力的方法学研究

Method for recording tension changes of small pulmonary artery and vein using the isolated vessel tension measurement system

目的 对大鼠离体血管环进行标准化处理,建立肺动、静脉离体血管环标准化实验方法,从而建立更科学的肺血管疾病体外实验研究模型.方法 SPF级雄性SD大鼠3只,麻醉后测定右心室压力,分离肺动、静脉并制备内径200~400μm及长2 mm的肺动、静脉血管环,用2根内径40μm的金属丝将血管环固定在浴槽中;以100μm/次逐步增加血管环内周长,使用DMT张力测定仪器和Acknowledge数据采集分析系统记录张力变化,做被动长度-张力指数曲线,根据测得的血管压力计算出相应的血管张力及螺旋测微尺的读数;调整初始张力并平衡30 min后用60 mmol/L高钾预刺激3次达最佳收缩活性,用去氧肾上腺素（PE）和乙酰胆碱检测肺动脉环的舒缩活性和内皮完整性,用血栓烷素类似物（U46619）和罂粟碱检测肺静脉环的舒缩活性.结果 使用1μmol/L PE刺激肺动脉环收缩达0.39 mg,加入10μmol/L乙酰胆碱舒张肺动脉环其舒张度为92%.使用1μmol/L U46619刺激肺静脉环收缩力达0.13 mg,加入10μmol/L罂粟碱舒张肺静脉环其舒张度为84%.结论 本研究成功制备舒缩活性良好的肺动、静脉血管环,为建立标准化的肺动、静脉离体血管环张力系统提供了基础.

Objective To establish a standardized method for isolated pulmonary artery and vein rings with different diameter , pressure and length , which could provide a more scientific method for in vitro study of pulmonary vessel diseases .Methods Male SD rats were anesthetized , and the right ventricular systolic pressure were measured .Small pulmonary artery and vein rings with 200-400μm in diameter and 2 mm in length were prepared by dissecting pulmonary arteries and veins .The pulmonary vessel rings were mounted in the organ bath by 2 stainless steel wires with diameter of 40μm.Then the internal circumference of the vessel rings was increased gradually with 100μm per step.At the same time the vascular tension was recorded by the Myograph System and Acknowledgement data acquisition system , and subsequently the passive length-tension exponential curve was made .The initial tension of the rings was set , equilibrated for another 30 min, and then stimulated with 60 mmol/L KCl 3 times, and the best contractile reactivity was achieved.The contractile reactivity of pulmonary artery rings and endothelial integrity were detected by exposure to 1 μmol/L phenylephrine （ PE ） and 10 μmol/L acetylcholine （ Ach ） , while the contractile reactivity of pulmonary vein rings was detected by exposure to 1 μmol/L U46619 and 10 μmol/L papaverine.Results The contraction and relaxation effects of the pulmonary artery rings reached 0.39 mg and 92%when they were stimulated by 1 μmol/L PE and 10 μmol/L Ach.The contraction and relaxation effects of pulmonary vein rings were up to 0.13 mg and 84%when they were exposed to 1 μmol/L U46619 and 10 μmol/L papaverine , respectively.Conclusion Pulmonary artery and vein rings with appropriate basal tension and optimal vasodilator activity were prepared , and a standardized method of tension experiment for isolated pulmonary artery and vein rings established.