【摘要】 目的:应用聚乙二醇(PEG)沉淀法提取关节液来源外泌体,并检测外泌体的形态及分子生物学特征,为关节液来源外泌体的进一步研究奠定基础。方法:用8%PEG从关节积液中分离、提取目标颗粒,然后经透射电子显微镜观察其形态,采用纳米颗粒示踪分析仪分析其大小及纯度,蛋白质印迹法检测其标记物蛋白的表达。结果:所获目标颗粒呈圆形或椭圆形囊泡,质膜完整,大小不一,直径约为35～155nm,且阳性表达CD9及CD63蛋白。结论:对目标颗粒的形态、大小及蛋白分析结果证实PEG沉淀法能够从关节积液中分离、提取外泌体,且操作方法简单,可为后续研究奠定基础。更多还原

【Abstract】 Objective:Exosomes in joint effusion were purified by polyethylene glycol(PEG)method,their morphology and molecular biological characteristics were detected,which laid a foundation for further research on exosomes from joint effusion.Methods:The target particles were separated and purified from the joint effusion by 8% PEG.The morphology was observed by transmission electron microscopy.The size and purity of the target particles were analyzed by nanoparticle tracer analyzer.The expression of the marker protein was detected by Western blotting.Results:The target particles were round or elliptical vesicles.The plasma membrane was intact and varied in size,with a diameter of about 35～155 nm,and positively expressed CD9 and CD63 proteins.Conclusion:The morphology,size and protein analysis of the target particles confirmed that the PEG precipitation method can successfully separate and purify exosomes from joint effusion,and the operation method is simple,which lays a good foundation for subsequent research.更多还原