Olson E B, Orbeck H, Graven S N, Zachman R D
J Appl Physiol Respir Environ Exerc Physiol. 1977 Sep;43(3):557-62. doi: 10.1152/jappl.1977.43.3.557.
To achieve an isolated, ventilated, perfused newborn rabbit lung preparation, newborn rabbits are anesthetized and ventilated. The pulmonary vasculature is perfused with a Krebs-Ringer bicarbonate solution containing glucose and plasma expanders, and the lungs are isolated in a 37 degrees C chamber at 100% humidity. The success of a perfusion is evaluated on three levels. 1) Visual. 2)mechanical: a, drop in flow of greater than 20% indicates failure; b, an increase of more than 25% of the initial lung weight indicates fluid accumulation and failure. 3) Biochemical: a, the endogenous ATP concentration after perfusion; b, the level of the enzyme choline phosphotransferase (CPT) after perfusion. Isolated, perfused, ventilated newborn rabbit lungs maintained for 4 h with no changes in the monitored physical and mechanical parameters have an endogenous ATP level of 1.06 +/- 0.06 (2 SD) mumol ATP/mg wet weight and a CPT level of 1.34 +/-0.15 (2 SD nmol [14C]CDP-choline per milligram protein in 30 min. These values are stable at a level lower than observed in intact, hypoxic, newborn rabbit's lungs. In constrast, if perfusion is maintained after any of the monitored criteria indicate failure, the endogenous ATP concentration and CPT activity are significantly depressed.
为制备出孤立、通气、灌注的新生兔肺标本,需对新生兔进行麻醉和通气。用含有葡萄糖和血浆扩容剂的 Krebs - Ringer 碳酸氢盐溶液灌注肺血管,然后将肺置于湿度为 100%、温度为 37 摄氏度的腔室内进行孤立处理。灌注成功与否从三个层面进行评估。1) 视觉层面。2) 力学层面:a,流量下降超过 20% 表明失败;b,肺初始重量增加超过 25% 表明有液体蓄积且失败。3) 生化层面:a,灌注后的内源性 ATP 浓度;b,灌注后的胆碱磷酸转移酶(CPT)水平。孤立、灌注、通气的新生兔肺在监测的物理和力学参数无变化的情况下维持 4 小时,其具有的内源性 ATP 水平为 1.06±0.06(2 个标准差)微摩尔 ATP/毫克湿重,CPT 水平为 1.34±0.15(2 个标准差)纳摩尔[14C]CDP - 胆碱/毫克蛋白质(30 分钟内)。这些值稳定在低于完整、低氧新生兔肺所观察到的水平。相反,如果在任何监测标准表明失败后仍维持灌注,内源性 ATP 浓度和 CPT 活性会显著降低。