持续灌注和静态保存对中性粒细胞凋亡的影响

Effect of Dynamic Perfusion and Static Preservation on Apoptosis of Neutrophil

为研究体外循环机持续灌注、间断压力灌注、单纯低温保存犬离体肺对支气管肺泡灌洗液中中性粒细胞凋亡变化以及对肺泡巨噬细胞吞噬凋亡中性粒细胞的影响,探索最佳的离体肺保存方式,将30只Mongrel犬随机分成3组,为体外循环组、压力灌注组和低温保存组,每组10只。在保持机械通气的条件下,完整摘取双肺,3种不同方式保存离体肺,并按时间点留取支气管肺泡灌洗液标本。流式细胞仪检测犬离体肺支气管肺泡灌洗液中中性粒细胞凋亡,免疫组织化学法检测犬离体肺肺泡巨噬细胞对凋亡中性粒细胞吞噬。随着犬肺离体时间的推移,各实验组离体肺支气管肺泡灌洗液中凋亡的中性粒细胞明显减少,在每个时间点上,体外循环组中性粒细胞的凋亡最多,压力灌注组中次之,低温保存组中最少。各实验组吞噬凋亡中性粒细胞的巨噬细胞随着离体时间的延长呈逐渐减少的趋势,在肺离体各时间点上,体外循环组中巨噬细胞对凋亡中性粒细胞的吞噬清除优于间断压力灌注组和单纯低温保存组。采用体外循环机持续灌注犬离体肺,能增加中性粒细胞凋亡,促使吞噬细胞吞噬凋亡的中性粒细胞,避免发生凋亡延迟,从而减轻离体肺组织的缺血再灌注损伤,具有较好的肺保护作用。

In order to study the effect of dynamic perfusion of extracorporeal circulation, intermittent pressure perfusion and simple hypothermic preservation of canine vitro lungs on changes of neutrophil apoptosis in BALF and alveolar macrophage phagocytosis of apoptotie neutrophils to explore the best way to preserve vitro lung, thirty Mongrel dogs were randomly divided into 3 groups： extracorporeal circulation perfusion group, intermittent pressure perfusion group and simple low temperature preservation group, and in each group there were 10 dogs. Both lungs were completely resected under the condition of keeping mechanical ventilation, which were preserved in three ways, and the samples of BALF were collected by time point. The apoptosis of the neutrophils in BALF in vitro lungs were assayed by flow cytometry and immunohistoehemistry was used to detect the macrophages phagocytosis of the apoptotic neutrophils in vitro lungs were detected by immunohistochemical method. With the pass of the time of canine lung in vitro, the apoptosis of the neutrophils in BALF showed an obvious trend of decline in each experimental group, and at each time point, the apoptosis of the neutrophils in extracorporeal circulation group was higher than pressure perfusion group, and that of low-temperature preservation group was the lowest. In each experimental group, the macrophages phagocytosis of apoptotic neutrophils displayed a trend of decline with the pass of time in vitro, and at the same time point, the macrophages phagocytosis of apoptotic neutrophils in extracorporeal circulation group was better than the other two groups. 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. The research showed that the extracorporeal circulation perfusion of canine vitro lungs could increase neutrophil apoptosis to promote the phagoeytosis of neutrophil phagocytosis of apoptotic cells and avoid the occurrence of delayed apoptosis so as to reduce the ischemia-reperfusion injury of isolated lung tissue and protect the lung.