输血相关急性肺损伤动物模型的建立及可溶性CD40配体在其发生机制中的作用

Establishment of An Animal Model of Transfusion-related Acute Lung Injury and the Role of Soluble CD40 Ligand in the Development of Transfusion-related Acute Lung Injury

目的建立更符合临床实际情况的输血相关急性肺损伤动物模型并在该动物模型中探究可溶性CD40配体(s CD40L)在输血相关急性肺损伤(TRALI)发生中的作用。方法采用创伤-失血-大量输血的方法建立TRALI动物模型,通过大鼠肺组织病理学改变、伊文斯兰染液漏出量、支气管肺泡灌洗液蛋白漏出量判断大鼠是否发生肺水肿以判断建模是否成功。检测大鼠红细胞储存前后及受血大鼠血浆中s CD40L含量以探讨s CD40L在TRALI发生中的作用。结果 7 d浓缩红细胞(PRBC)组大鼠肺组织病理学改变示肺泡上皮细胞增生、肺泡间隔增厚及间质大量炎性细胞浸润。7 d PRBC组大鼠支气管肺泡灌洗液蛋白漏出量(13.17±5.76)mg明显高于正常对照组(1.21±0.66)mg及生理盐水(NS)对照组(4.94±2.15)mg(F=17.605,P<0.001)。7 d PRBC组大鼠伊文斯兰染液漏出量(0.0109±0.0067)%/min显著高于NS对照组(0.0026±0.0006)%/min(t=2.998,P=0.03)。储存7 d PRBC中s CD40L含量(451.58±73.28)pg/ml显著高于0 d PRBC(277.94±98.18)pg/ml(t=2.834,P=0.03)。7 d PRBC组大鼠血浆s CD40L含量(878.21±125.30)pg/ml明显高于正常对照组(289.78±62.60)pg/ml和NS对照组(418.07±47.68)pg/ml(F=78.715,P<0.001)。结论采用创伤-失血-大量输血的方法成功建立大鼠TRALI模型,接受大量输血的TRALI大鼠血浆s CD40L含量明显高于正常对照组及NS对照组,提示s CD40L可能在TRALI的发生中发挥一定作用。

Objective To establish an animal model of transfusion-related acute lung injury(TRALI)and investigate the role of soluble CD40 ligand(sCD40L)in the development of TRALI.Methods The TRALI animal model established by trauma-hemorrhage-transfusion.Lung edema was evaluated by histopathological examination and the protein and Evans blue dye accumulation in bronchoalveolar lavage fluid.The concentration of sCD40L in storage packed red blood cell(PRBC)and rat’s plasma was measured by enzyme-linked immunosorbent assay(ELISA).Results There were obvious epithelial hyperplasia and inflammatory cell infiltration in the lung tissue of 7 d-PRBC-treated group.The accumulation of protein in bronchoalveolar lavage fluid of 7 d-PRBC-treated group[(13.17±5.76)mg]was significantly higher than that in normal controls[(1.21±0.66)mg]and normal saline(NS)-treated group[(4.94±2.15)mg](F=17.605,P<0.001).The leakage amount of Evans blue dye in 7 d-PRBC-treated group[(0.0109±0.0067)%/min]was significantly higher than that in NS-treated group[(0.0026±0.0006)%/min](t=2.998,P=0.03).The concentration of sCD40L of the 7 d PRBC[(451.58±73.28)pg/ml]was significantly higher than 0 d PRBC[(277.94±98.18)pg/ml](t=2.834,P=0.03).The concentration of sCD40L in the plasma of 7 d-PRBC-treated group[(878.21±125.30)pg/ml]was significantly higher than those in normal controls[(289.78±62.60)pg/ml]and NS-treated group[(418.07±47.68)pg/ml](F=78.715,P<0.001).Conclusions The TRALI animal model was successfully established with trauma-hemorrhage-transfusion.The concentration of sCD40L in plasma of rats with massive transfusion is remarkably increased,suggesting sCD40L may play a role in the development of TRALI.