【摘要】 目的分析间断灌注保存中灌注容量对离体猪心冠状动脉血管舒缩因子和炎性介质的影响,探寻最佳的移植供心血管保存方法。方法以巴拿马小香猪为对象,每组6头。根据保存技术不同分为浸泡保存组和灌注保存组(每隔2 h间断灌注一次,灌注容量分别为50、100、150、200、250 ml)。灌注液为30μmol/L二氮嗪强化Celsior液,保存温度为4℃,时间10 h。通过免疫组化和RT-PCR方法分别从蛋白水平和基因分子水平上比较各组间冠状动脉血管中内皮型一氧化氮合成酶(e NOS)、内皮素1(ET-1)、细胞黏附分子1(ICAM-1)、血管性血友病因子(v WF)表达情况和电镜血管超微结构。结果 e NOS蛋白和mRNA表达在浸泡保存组最低,灌注保存组中随着灌注容量的增多而增高;ET-1、ICAM-1、v WF蛋白和mRNA表达在浸泡保存组最高,灌注保存组中随着灌注容量的增多而降低;其中200 ml灌注组和250 ml灌注组各指标除ICAM-1蛋白外免疫组化测定值差异均无统计学意义;电镜超微结构上均较好地保留内皮细胞相对完整的覆盖以及较少的空泡出现,且内皮下层较厚。结论在长时间心脏保存中,间断灌注保存能更好地保护冠状动脉血管舒缩因子,减少血管炎性介质的释放,内皮结构保存较完整;提示间断灌注保存技术对冠状动脉血管的保护作用优于单纯低温浸泡保存技术;且间断2 h灌注一次,每次灌注容量为200 ml的间断灌注保存技术较优越。更多还原

【Abstract】 Objective To compare the protective effect of different perservation on coronary artery in isolated heart of pigs. Methods A total of 36 Bama-miniature-pigs’ donor hearts were arrested with Celsior cardioplegia solution enhanced with Diazoxide( DE,30 μmol / L). Isolated hearts were stored in this solution in immersion storage group or place in a device that providing intermittent perfusion with 50,100,150,200,250 ml solution respectively every 2 hours in the intermittent perfusion group at 4 ℃ for 10 hours( n = 6 per group). After storage,left anterior descending coronary artery samples were frozen,extracted,and analyzed the endothelial nitric oxide synthase( e NOS),endothelin-1( ET-1),intercellular adhesion molecule-1( ICAM-1),von Willebrand factor( v WF) and ultrastructure by immunohistochemisty and RT-PCR. Results The expression of e NOS protein and mRNA in the immersion preservation group was minimum,in perfusion preservation group,e NOS protein and mRNA increased with the increasing of capacity; the expression of ET-1,ICAM-1,v WF protein and mRNA were highest in immersion preservation,in perfusion preservation group,they increased with the reducing of capacity; in 200 ml and250 ml perfusion group,there was no significant difference in indexes except ICAM-1 protein. Both group retained relatively complete coverage better endothelial cells ultrastructure and fewer vacuoles and subendothelial layer was thicker. Conclusion These results indicate that the intermittent perfusion is optimal means on coronary artery in isolated heart preservation with the support of Celsior cardioplegia solution enhanced with DE( 30 μmol / L). They can better protect the coronary vasomotor factors,reduce vascular inflammatory mediators released and protect coronary artery endothelial cells’ structure. And it is more superior protection when the heart is perfused 200 ml solution every 2 hours.更多还原