体外循环对血浆内皮细胞来源外泌体含量及外泌体中热休克蛋白表达的影响  

Influence of cardiopulmonary bypass on the content of plasma endothelium-derived exosomes and the expression of heat shock protein 70 in exosomes

作　　者：朱耀斌[1] 李志强[1] 丁楠[1] 伊寒露 沈磊[1] 赵宇东[1] 叶赞凯[2] 刘扬[2] 樊星[2] ZHU Yao-bin;LI Zhi-qiang;DING Nan;YIN Han-lu;SHEN Lei;ZHAO Yu-dong;YE Zan-kai;LIU Yang;FAN Xing(Pediatric Cardiac Surgery, Beijing Children＇s Hospital Affiliated to Capital Medical University , National Children＇s Medical Center, Beijing 100045, China)

机构地区：[1]国家儿童医学中心首都医科大学附属北京儿童医院小儿心脏外科,北京100045 [2]首都医科大学附属北京安贞医院小儿心脏中心,北京100029

出　　处：《中华实用诊断与治疗杂志》2018年第5期437-439,共3页Journal of Chinese Practical Diagnosis and Therapy

基　　金：北京市优秀人才培养资助(2014000021469G233);北京市卫生系统高层次卫生技术人才-学科骨干(2015-3-048);北京市卫生系统高层次卫生技术人才-学科骨干(2015-3-051);北京市卫生系统高层次卫生技术人才-学科骨干(2014-3-043);北京市自然科学基金(7152045);北京市自然科学基金(7184204);北京市医管局培育计划(PX2016046);北京市科学技术委员会首都临床特色应用研究(Z171100001017048);国家自然科学基金(81371443);国家自然科学基金(81400305)

摘　　要：目的探讨体外循环(cardiopulmonary bypass,CPB)下行先天性心脏畸形矫治术患儿血浆内皮细胞来源外泌体含量及外泌体中热休克蛋白(heat shock protein 70,HSP70)表达变化。方法先天性心脏畸形患儿60例,均于CPB下行先天性心脏畸形矫治术,分别于CPB前及再灌注1、6、12h采集桡动脉血,采用双抗体夹心法检测血浆心肌肌钙蛋白T(cardiac troponin,cTnT)水平,采用密度梯度离心法分离血浆中内皮细胞来源外泌体,Western blot法检测外泌体中HSP70表达。结果 CPB前内皮细胞来源外泌体直径40~100nm,呈椭圆形或圆形;再灌注6h,内皮细胞来源外泌体直径、形态与CPB前无明显变化;再灌注1、6、12h,血浆cTnT水平[(1.347±0.215)、(1.648±0.235)、(1.254±0.201)μg/L]、内皮细胞来源外泌体含量[(167.14±26.54)×10~6微粒/mL、(210.34±31.20)×10~6微粒/mL、(189.64±26.45)×10~6微粒/mL]、外泌体中HSP70表达(0.76±0.12、0.85±0.14、0.73±0.09)均明显高于CPB前[(0.014±0.001)μg/L、(76.34±13.20)×10~6微粒/mL、0.18±0.03](P<0.05);再灌注6h时各指标均高于再灌注1、12h(P<0.05),再灌注1h与再灌注12h时各指标比较差异均无统计学意义(P>0.05)。结论 CPB可增加行CPB下先天性心脏畸形矫治术患儿血浆中内皮细胞来源外泌体含量及外泌体中HSP70表达。Objective To investigate the influence of cardiopulmonary bypass（CPB）on the content of plasma endothelium-derived exosomes and the expression of heat shock protein 70（HSP70）in exosomes in patients receiving congenital heart malformations correction surgery. Methods Sixty children with congenital heart malformations underwent correction under CPB.The radial artery blood samples were collected at the time points of before CPB,and in1,6 and 12 hafter reperfusion respectively.The plasma level of cardiac troponin T（cTnT）was detected by double antibody Sandwich method.The plasma endothelial-derived exosomes were separated by density gradient centrifugation.The expression of HSP70 protein in exosomes was determined by Western blot.Results The endothelial-derived exosome before CPB was from 40 to 100 nm in diameter,in oval or round.There were no significant change in the diameter and morphology of endothelial-derived exosomes in 6 hafter reperfusion in comparison with those before CPB.The cTnT levels（（1.347±0.215）,（1.648±0.235）,（1.254±0.201）μg/L）,the contents of endothelial-derived exosomes（（167.14±26.54）×106 particles/mL,（210.34±31.20）×10^6 particles/mL,（189.64±26.45）×10^6 particles/mL）,and the expressions of HSP70 in exosomes（0.76±0.12,0.85±0.14,0.73±0.09）in 1,6 and 12 hafter reperfusion were significantly higher than those before CPB（（0.014±0.001）μg/L,（76.34±13.20）×10^6 particles/mL,0.18±0.03）（P〈0.05）,higher in 6 hafter reperfusion than those in 1 and 12 hafter reperfusion（P〈0.05）,and there were no significant differences in 1 hafter reperfusion in comparison with those in 12 hafter reperfusion（P〈0.05）.Conclusion CPB can increase the content of endothelium-derived exosomes in plasma and the expression of HSP70 in exosomes in children receiving congenital heart malformations correction surgery.

关 键 词：先天性心脏畸形 体外循环 先天性心脏畸形矫治术 外泌体 热休克蛋白70 

分 类 号：R726.5[医药卫生—儿科]