动态灌注与静态保存对犬离体肺组织中AQP1表达的影响

Effect of dynamic perfusion and static preservation on expression of AQP1 in vitro dog lung tissue

目的通过比较体外循环机持续灌注、间断压力灌注、单存低温保存离体肺的水通道蛋白1(AQP1)表达变化,探索离体肺保存的最佳保存方法。方法将30只Mongrel犬分成体外循环组、压力灌注组、低温保存组,每组10只。在保持机械通气的条件下,完整摘取双肺。进行体外循环机灌注、间断压力灌注、单存灌注离体肺循环后低温浸泡保存,并按时间点留取标本;HE染色法观察犬离体肺组织形态学改变,免疫组织化学法、蛋白免疫印迹法(Western blot)检测犬离体肺组织AQP1的表达。结果 HE染色发现,随着离体时间的延长,肺泡组织结构逐渐塌陷,炎性细胞及渗出物增多,肺泡间隔也逐渐增宽,肺泡腔内可见渗出物;在同一时间点,体外循环组的组织结构损伤变化比压力灌注组和低温保存组轻。在各实验组随着时间推移AQP1的表达量呈现下降趋势;在每个时间点上,体外循环组AQP1的表达量高于压力灌注组,压力灌注组又高于低温保存组。结论体外循环机械灌注对离体肺的保护作用显著优于间断压力灌注和单存低温保存。

Objective By comparing the change of expression of AQPI in vitro lung preserved by the continuous infusion of a heart-lung machine,continuous pressure perfusion and single low temperature,to explore the best method of vitro lung preservation. Methods Thirty Mongrel dogs were randomly divided into 3 groups,and both lungs were completely resected under the condition Of keeping mechanical ventilation. The vitro lungs were preserved by the way of the continuous infusion of a heart lung machine,continuous pressure perfusion and single low temperature,and collecting specimens according to the time point. HE staining was used to observe the morphological changes of vitro lung tissue. Immunohistochemistry and Western blot were used to detect the expression of AQP1 in vitro lung . Results HE staining found that as the time went by alveolar structure gradually collapsed,in flammatory cells increased,alveolar interval also gradually broadened and exudation could be seen in the alveolar cavity;at the same time point,organization structure of extraeorporeal circulation group changed lighter than pressure perfusion group and low-temperature preservation group. In each experimental group, the expression of AQP1 showed a trend of decline;at each time point, the expression of AQP1 in extracorporeal circulation group was higher than pressure perfusion group, and pressure infusion group was higher than that of low-temperature preservation group. Conclusion The protective effect of the continuous infusion of a heart-lung machine on vitro lung was better than continuous pressure perfusion and single low temperature.