Characteristics Of Plasma Exosomes In Rheumatoid Arthritis And Its Effect On The Function Of Human Vascular Endothelial Cells

Objective: Plasma-derived exosomes was extracted and identified in patients with rheumatoid arthritis.Methods: 1.The collection meets the 2010 American Rheumatism Association(ACR)/European Union against Rheumatism(EULAR)standards,patients with DAS28 score greater than 6 and normal,while excluding tumors,tuberculosis,suppurative arthritis,SLE 、 sjogren’s syndrome,myasthenia gravis,viral hepatitis,and other autoimmune diseases.2.the exosomes was isolated with the kit,the morphological characteristics,the size of the nanoparticle analyzer(NTA)were observed by transmission electron microscopy and the surface marker protein was identified by Western blotting.3.cultured human umbilical vein endothelial cells,the endothelial cell activity was detected by CCK8 cell proliferation test.4.the exosomes and PBS from plasma of patients and normal subjects were treated on HUVECs cells,which were divided into PBS,con-exo and ra-exo groups.5.fluorescence quantitative PCR was used to detect the changes of hypoxia inducible factor 1 a(hif-1)and vascular endothelial growth factor(VEGF)in cells.Results: The exosomes was double concave and disc-shaped cystic vesicles,and the diameter of the vesicles was about 50-150 nm.The flotillin1 and CD81,molecular weights were 47 KDa and 26 KDa respectively.compared with normal human con-exo,rheumatoid arthritis ra-exo significantly inhibited endothelial cell proliferation,migration and invasion,and the ability to secrete HIF-1a 、 VEGF cytokines was also significantly reduced,and the difference was statistically significant(P <0.05).Conclusion:the plasma exosomes from patients with rheumatoid arthritis has been successfully extracted.The plasma exosomes from patients with RA can inhibit vascular regeneration and may play an important role in RA complicated with vascular injury.