亚甲蓝光化学法病毒灭活血浆外泌体微小RNA差异表达谱分析 / 中国生物制品学杂志

ABSTRACT

@#Objective To explore the effect of methylene blue photochemistry(MB-P)viral inactivation treatment on the expression of microRNA(miRNA)in plasma exosomes,in order to provide a new reference for the quality control of MB-P virus inactivated plasma. Methods Whole blood samples of 11 healthy volunteers were collected from July 2021 to April2022. Fresh plasma from the same person was prepared into two parts,fresh frozen plasma(FFP)and MB-P virus inactivated plasma,respectively. The plasma exosomes were isolated by differential centrifugation,and identified by transmission electron microscopy(TEM)and nanoparticle tracking analysis(NTA). Then the expression profiles of miRNA were detected by microarray technique. Furthermore,four differentially expressed miRNA were verified by qRT-PCR,the target genes of differentially expressed miRNA were predicted by bioinformatics methods,and GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the differentially expressed genes. Results The morphological characteristics and diameters of the extracted vesicles of the two groups were consistent with the characteristics of exosomes. Compared with the control group,there were 14 differentially expressed miRNA in plasma exosomes of MB-P group,of which the expression of six miRNA was up-regulated and eight miRNA was down-regulated. The results of qRT-PCR were generally consistent with the expression trend of microarray. The target genes of differentially expressed miRNA were mainly involved in DNA binding,ion binding,catalytic activity and other functions,and participated in a variety of biological processes such as nucleic acid metabolism,biosynthesis,and transcription regulation. In addition,significantly enriched functional pathways were closely related to viral infectious diseases,tumors,PI3K-Akt signaling pathway,MAPK signaling pathway and so on.Conclusion The expression of exosome miRNA in MB-P virus inactivated plasma was different from that in FFP. The plasma exosome miRNA may be used as a potential reference for the quality evaluation of MB-P virus inactivated plasma.