炎症环境下巨噬细胞摄取软骨细胞来源外泌体的作用研究  

Study on Uptake of Exosomes Secreted by Chondrocytes by Macrophages in an Inflammatory Environment

作　　者：刘昱江 林才渊 刘金柱 李志鹏 夏天卫 沈计荣 LIU Yujiang;LIN Caiyuan;LIU Jinzhu;LI Zhipeng;XIA Tianwei;SHEN Jirong(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属医院骨伤科,南京210029

出　　处：《中国中医骨伤科杂志》2022年第4期1-4,9,共5页Chinese Journal of Traditional Medical Traumatology & Orthopedics

摘　　要：目的:在炎症环境下,探究巨噬细胞对软骨细胞分泌携载lncRNA PVT1的外泌体的摄取情况。方法:提取、培养大鼠软骨细胞和巨噬细胞,分别进行体外培养,通过透射电镜观察软骨细胞分泌的外泌体形态,通过Western Blot检测外泌体特异性蛋白CD9和CD63的表达。进而实验分为3组,对照组、低浓度软骨细胞分泌的外泌体+单核细胞培养组、高浓度软骨细胞分泌的外泌体+单核细胞培养组,通过RT-qPCR检测外泌体中lncRNA PVT1的表达情况,通过免疫荧光法检测巨噬细胞对软骨细胞分泌的外泌体的摄取情况。结果:在炎症环境下,软骨细胞能够分泌携载lncRNA PVT1的外泌体,外泌体特异性蛋白CD63和CD9的表达为阳性。免疫荧光法证实巨噬细胞可摄取该外泌体,且高浓度外泌体组细胞摄取外泌体的量较低浓度外泌体组多。结论:在膝骨关节炎的炎症环境下,巨噬细胞可摄取软骨细胞来源的携载lncRNA PVT1的外泌体。Objective:To investigate the uptake of exosomes carrying lncRNA PVT1 secreted by chondrocytes in macrophages in inflammatory environment.Methods:Rat chondrocytes and macrophages were isolated and cultured in vitro.The exocrine morphology of chondrocytes was observed by transmission electron microscope,and the expression of CD9 and CD63 were detected by Western Blot.Further experiments were divided into three groups:control group,low concentration chondrocytes + monocytes culture group and high concentration chondrocytes + monocytes culture group.The expression of lncRNA PVT1 in exosomes was detected by RT-qPCR,and the uptake of exosomes secreted by chondrocytes by macrophages was detected by immunofluorescence.Results:Chondrocytes could secrete the exosomes carrying lncRNA PVT1 in inflammatory environment,and the expression of exosome specific protein CD9 and CD63 was positive.Macrophages could ingest the exosomes in inflammatory environment,and the cells in the high concentration group ingested more exosomes than those in the low concentration group.Conclusion:Macrophages can ingest the exosomes containing lncRNA PVT1 secreted by chondrocytes in inflammatory environment.

关 键 词：膝关节骨性关节炎 炎症 巨噬细胞 外泌体 长链非编码RNA 

分 类 号：R-33[医药卫生]