FTY720对输血相关急性肺损伤的作用机制初探  

Effect of FTY720 on transfusion-related acute lung injury:a preliminary investigation

作　　者：周亚玲 李玲[1,2] 田力 王珏 何芮[3] 刘忠 ZHOU Yaling;LI ling;TIAN Li;WANG Jue;HE Rui;LIU Zhong(Institute of Blood Transfusion,Chinese Academy of Medical Sciences&Peking Union Medical College,Chengdu 610052,China;Key Laboratory of Transfusion Adverse Reactions,Chinese Academy of Medical Sciences;Department of Blood Transfusion,The First Affiliated Hospital of Anhui Medical University)

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052 [2]中国医学科学院输血不良反应研究实验室 [3]安徽医科大学第一附属医院输血科

出　　处：《中国输血杂志》2023年第5期403-410,共8页Chinese Journal of Blood Transfusion

基　　金：中国医学科学院医学与健康科技创新工程(2021-1-I2M-060);四川省国际科技创新合作/港澳台科技创新合作项目(2020YFH0024)。

摘　　要：目的探讨不同剂量芬戈莫德(Fingolimod,FTY720)对非抗体介导的输血相关急性肺损伤(transfusion-related acute lung injury,TRALI)的作用及机制。方法采用脂多糖(lipopolysaccharide,Lps)预刺激,不同保存天数的血小板(platelet,Plt)进行二次打击的方法构建TRALI小鼠模型,通过肺组织匀浆中蛋白浓度、髓过氧化物酶(myeloperoxidase,Mpo)活性、肺组织湿干重比(wet/dry weight ratio,W/D比值)、肺组织损伤评分及病理切片判断建模是否成功,检测不同保存天数血小板中的神经酰胺和鞘氨醇-1-磷酸(sphingosine-1-phosphate,S1P)含量。LPS注射1h后给予不同剂量的FTY720以探讨FTY720在TRALI中的作用。通过WB分析血管内皮钙黏蛋白(vascular endothelial cadherin,VE-cadherin)、密闭蛋白1(zonula occludens-1,ZO-1)的表达水平以研究FTY720的作用机制。结果输注保存5 d Plt(d5 Plt组)的小鼠表现出TRALI的典型症状,肺组织匀浆蛋白浓度(6546.38±409.50)μg/mL、MPO活性(49.38±4.43)U/L、W/D比值4.79±0.21、肺组织损伤评分7.24±0.38与其余各组的差异均具统计学意义(P<0.05)。随着血小板保存时间的增加,神经酰胺含量逐渐升高,S1P含量逐渐降低,两者的比例失衡。d5 Plt的神经酰胺含量(58.37±5.69)μmol/L、S1P含量(149.81±4.86)nmol/L与其余各组的差异均具统计学意义(P<0.01)。FTY720预防给药后,1 mg/kg FTY720对TRALI小鼠无明显作用,其肺组织匀浆蛋白浓度(6170.26±545.50)μg/mL、MPO活性(45.97±4.79)U/L、W/D比值4.88±0.25、肺组织损伤评分7.92±0.65均显著高于正常组和LPS对照组(P<0.01)。低剂量(0.5、0.2、0.1mg/kg)FTY720组则可缓解肺损伤,其蛋白浓度、MPO活性、W/D比值、肺组织损伤评分均显著低于d5 Plt组(P<0.05)。病理切片也呈现相似的结果。在内皮细胞间连接蛋白表达方面,1 mg/kg FTY720组的VEcadherin表达水平显著低于正常组和LPS对照组(P<0.05),低剂量(0.5、0.2、0.1mg/kg)FTY720组VE-cadherin、ZO-1的表达水Objective To investigate the effects and mechanisms of different doses of fingolimod(FTY720)on non-antibody-mediated transfusion-related acute lung injury(TRALI).Methods A TRALI mouse model was constructed using lipopolysaccharide(LPS)pre-stimulation and platelets(Plt)of different storage days for second strike.The success of the modeling was determined by protein concentration in lung tissue homogenates,myeloperoxidase(MPo)activity,lung wet/dry weight ratio(W/D ratio),lung tissue damage score and pathological sections.Ceramide and sphingosine-1-phosphate(S1P)contents in platelets of different storage days were detected.FTY720 was administered 1 h after LPS injection to investigate the role of FTY720 in TRALI.The expression levels of vascular endothelial cadherin(VE-cadherin)and zonula occludens-1(ZO-1)were analyzed by WB.Results Mice infused with stored 5-day Plt(d5Plt group)exhibited typical signs of TRALI,and the differences in lung tissue homogenate protein concentration(6546.38±409.50)μg/mL,MPO activity(49.38±4.43)U/L,W/D ratio 4.79±0.21,and lung tissue damage score 7.24±0.38 from the rest of the groups were statistically significant(P<0.05).With the increase of platelet storage time,the ceramide content gradually increased and S1P content gradually decreased,and the ratio of the two was imbalanced.d5Plt showed statistically significant differences(P<0.01)in ceramide content(58.37±5.69)μmol/L and S1P content(149.81±4.86)nmol/L from the rest of the groups.After preventive administration of FTY720,1 mg/kg FTY720 had no significant effect on TRALI mice,whose lung tissue homogenate protein concentration(6170.26±545.50)μg/mL,MPO activity(45.97±4.79)U/L,W/D ratio 4.88±0.25,and lung tissue damage score 7.92±0.65 were significantly higher than those of the normal and LPS control groups(P<0.01).The low-dose(0.5,0.2,and 0.1 mg/kg)FTY720 group alleviated lung injury,and its protein concentration,MPO activity,W/D ratio,and lung tissue injury score were significantly lower than those of the d5Plt group(P<0.0

关 键 词：FTY720 输血相关急性肺损伤 神经酰胺 S1P 

分 类 号：R457.13[医药卫生—治疗学]