利用实时能量代谢检测技术研究贮存单采血小板的线粒体呼吸功能  

Study on mitochondrial respiratory function in stored apheresis platelets using real-time energy metabolism detection technology

作　　者：赵明 张琛 朱立国 ZHAO Ming;ZHANG Chen;ZHU Liguo(Department of Blood Transfusion,Qilu Hospital(Qingdao),Cheeloo College of Medicine,Shandong University,Qingdao 266035,China;Department of Medical Experimental Center,Qilu Hospital(Qingdao),Cheeloo College of Medicine,Shandong University,Qingdao 266035,China)

机构地区：[1]山东大学齐鲁医院(青岛)输血科,山东青岛266035 [2]山东大学齐鲁医院(青岛)医学实验中心,山东青岛266035

出　　处：《中国输血杂志》2024年第9期1013-1017,1041,共6页Chinese Journal of Blood Transfusion

基　　金：山东省自然科学基金(ZR2021QH345);山东大学齐鲁医院(青岛)院内科研基金(QDKY2020BS07)。

摘　　要：目的探索利用实时能量代谢检测技术检测单采血小板(AP)中线粒体呼吸功能(MRF)的最佳条件参数,并研究AP在贮存过程中MRF的变化。方法收集AP标本,利用Seahorse XF检测其中MRF,通过调整AP的上样浓度、洗涤条件、线粒体呼吸调节药物浓度等多项参数,最后分别研究贮存1 d和5 d的AP中MRF。结果AP上样浓度为5×10^(7)/100μL时,基础呼吸值316±65为最优;洗涤组的基础呼吸值329±120比未洗涤组的57±7更优(P<0.05);FCCP浓度以2μmol/L为最优。贮存1 d AP的基础呼吸值为683±161,而贮存5 d AP的基础呼吸值为140±23(P<0.05);贮存1 d AP的最大呼吸值为1044±82,而贮存5 d AP最大呼吸值<200(P<0.05)。2组的MRF动力曲线有明显差异。结论实时能量代谢检测技术可以实现对AP中MRF的检测。AP在贮存过程中MRF明显受损,提示这可能是血小板贮存损伤的重要新机制。Objective To explore the optimal parameters for detecting mitochondrial respiratory function(MRF)in apheresis platelets(AP)using real-time energy metabolism detection technology,and to study the change of MRF in AP during storage.Methods AP samples were collected and detected for MRF using the Seahorse XF device.Multiple parameters such as AP loading concentrations,washing conditions and mitochondrial respiratory regulatory drug concentrations were adjusted.Finally,the MRF in AP stored for 1 and 5 days were studied.Results The optimal parameters for detecting MRF in AP were as follows:The loading concentration was 5×10^(7)/100μL and basal respiration was 316±65;Basal respiration in washing group was 329±120 compared with 57±7 in unwashing group;The concentration of FCCP was 2μmol/L.The basal respiration in AP for 1 day was 683±161,while was 140±23 for 5 days,P<0.05;The maximum respiration in AP for 1 day was 1044±82,while was less than 200 for 5 days,P<0.05.There was a significant difference in MRF dynamic curves between the two groups.Conclusion The real-time energy metabolism detection technology was able to detect MRF in AP effectively.The MRF significantly damaged during AP storage,indicating that it may be one new mechanism of platelet storage lesion.

关 键 词：单采血小板 线粒体 呼吸功能 实时能量代谢检测 贮存 

分 类 号：R457.1[医药卫生—治疗学] R331.143[医药卫生—临床医学]