内皮前体细胞来源外泌体减轻新生大鼠肺泡Ⅱ型上皮细胞高氧损伤的研究  

The alleviating effects of endothelial progenitor cells-derived exosomes on hyperoxia-induced injury of typeⅡalveolar epithelial cells in neonatal rats

作　　者：曾雯 毛劲 胡旭红 高淑强 巨容 Zeng Wen;Mao Jin;Hu Xuhong;Gao Shuqiang;Ju Rong(Department of Neonatology,Chengdu Women's&Children's Central Hospital,School of Medicine,University of Electronic Science and Technology of China,Chengdu 611731,China)

机构地区：[1]电子科技大学医学院附属妇女儿童医院·成都市妇女儿童中心医院新生儿科,成都611731

出　　处：《中华新生儿科杂志（中英文）》2022年第5期452-456,共5页Chinese Journal of Neonatology

基　　金：国家自然科学基金(81401247);四川省科技厅应用基础研究(2019Y30648)。

摘　　要：目的探讨内皮前体细胞(endothelial progenitor cell,EPC)来源外泌体对高氧诱导的新生大鼠肺泡Ⅱ型上皮细胞(typeⅡalveolar epithelial cells,AECⅡ)损伤的干预作用。方法使用全外泌体分离试剂盒从大鼠EPC培养液中提取外泌体,将体外培养的新生大鼠AECⅡ分为对照组、高氧组、干预组。对照组为空气+5%CO_(2)培养,高氧组为95%O_(2)+5%CO_(2)培养,干预组培养基中加入0.1 mg/ml EPC来源外泌体,并予95%O_(2)+5%CO_(2)培养。于2、4、6 d使用细胞增殖及毒性检测试剂盒检测AECⅡ活力、流式细胞仪检测AECⅡ凋亡。结果透射电镜观察EPC培养上清中分离的外泌体均符合外泌体的形态学特征。Dil标记EPC来源外泌体后与AECⅡ共培养24 h,在荧光显微镜下可观察到AECⅡ胞浆中有Dil荧光,提示EPC来源外泌体可以被AECⅡ摄取进入胞浆。与对照组相比,高氧组AECⅡ活力受损,细胞凋亡增加,并且损伤程度随高氧时间延长而加重(P<0.001)。EPC来源外泌体干预组细胞损伤程度低于高氧组(P<0.001)。与对照组相比,干预组在高氧4 d、6 d时的细胞活力低于对照组(P值分别为0.029、0.005),在高氧6 d时的细胞凋亡高于对照组(P=0.007)。结论EPC来源外泌体干预可减轻高氧诱导对AECⅡ的损伤,但不能完全逆转损伤。Objective To study the effects of endothelial progenitor cells(EPC)-derived exosomes on hyperoxia-induced injury in typeⅡalveolar epithelial cell(AECⅡ)in neonatal rats.Methods EPCs of rats were cultured and exosomes were collected using Total Exosome Isolation kit.Primary cultured AECⅡof neonatal rats were randomly assigned into three groups:the control group,the hyperoxia group and the exosome group.The control group was cultured in room air with 5%CO_(2),the hyperoxia group was cultured in 95%O_(2) with 5%CO_(2) and the exosome group was cultured with 0.1 mg/ml EPC-derived exosomes in 95%O_(2) with 5%CO_(2).Cell viability was detected using cell counting kit-8(CCK-8)and apoptosis was detected using flow cytometry on d2,d4,and d6.Results EPC-derived exosomes isolated from EPC culture supernatant were confirmed morphologically using transmission electron microscopy.After co-incubation of Dil-labeled EPC-derived exosomes with AECⅡfor 24 h,Dil fluorescence was detected in the cytoplasm of AECⅡ,indicating exosomes were uptaken by AECⅡ.Compared with the control group,hyperoxia decreased cell viability and increased apoptosis of AECⅡand the injury was aggravated with the prolongation of hyperoxia duration(P<0.001).Cell injury in the exosome group was milder than the hyperoxia group(P<0.001).Compared with the control group,cell viability on d4 and d6 of hyperoxia was lower(P=0.029 and 0.005 respectively)and cell apoptosis at d6 of hyperoxia was higher in the exosome group(P=0.007).Conclusions EPC-derived exosomes may partially attenuate hyperoxia-induced cell injury in neonatal rat AECⅡ.

关 键 词：内皮前体细胞 外泌体 肺泡Ⅱ型上皮细胞 高氧 凋亡 

分 类 号：R363[医药卫生—病理学]