Coagulation reaction of platelet extracellular vesicles in critically burned patients based on proteomics technology
Objective To explore the differential proteins related to coagulation reaction in platelet extracellular vesicles(PLT-EVs)between critically burned patients and healthy volunteers by proteomics technology.Methods A total of 6 healthy volunteers and 6 patients with severe burns in the Third Affilia-ted Hospital of Inner Mongolia Medical University were selected,and venous blood was extracted.PLT-EVs were obtained by high-speed centrifugation,and identified by transmission electron microscopy,Nanoparti-cle Tracking Analysis and Western blotting.The proteomic techniques were used to analyze and quantify two groups of PLT-EVs proteins.The differential proteins were screened with a differential multiple of 1.5(up-regulated or down-regulated).Differentially expressed proteins were analyzed through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.The top 6 proteins related to co-agulation differences were screened and analyzed.The measurement data in line with the normal distribu-tion were expressed as mean±SD,and the independent sample t test was used to compare the mean value of the two groups.P＜0.05 was considered to be statistically significant difference.Results Under the electron microscope,the PLT-EVs showed typical double-layer membrane.The TSG101,CD63,CD9 and CD81 were identified positively by Western blotting.The average particle sizes of the PLT-EVs were 168 nm and 153.5 nm respectively.A total of 853 proteins in PLT-EVs were identified by quantitative analysis,with 191 proteins being up-regulated and 77 proteins being down-regulated.The GO and KEGG analysis showed differential proteins were mainly involved in biological processes,such as coagulation func-tion,inflammatory response,immune response and body repair regulation.The top 6 proteins related to co-agulation reaction were plasminogen,dedicator of cytokinesis protein 5,rho-associated protein kinase 1,rap1 GTPase-activating protein 2,GRB2-related adapter protein 2 and protein kinase C alpha type.Conclusion PLT-EVs differential protein analysis could provide important theoretical basis for the regula-tion mechanism of platelet function and coagulation reaction in critically burned patients.
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