Objective:To explore the possible mechanism of neutrophil exosomes regulating macrophage apoptosis.Methods:neutrophils were induced by lipopolysaccharide,inflammatory factors were detected by ELISA,the morphology of e...Objective:To explore the possible mechanism of neutrophil exosomes regulating macrophage apoptosis.Methods:neutrophils were induced by lipopolysaccharide,inflammatory factors were detected by ELISA,the morphology of exosomes was identified by electron microscope,and the expression of mir-15a-5p in exosomes was detected by RT-PCR;Raw267.4 macrophages was treated with neutrophil exosomes and mir-15a-5p mimic respectively,CCK8 to detect cell viability,flow cytometry to detect apoptosis;The binding sites of mir-15a-5p and BCL2L2 were predicted and verified by double luciferase experiment;RT-PCR and Western blot verified that mir-15a-5p regulate the expression of BCL2L2.Results:lipopolysaccharide induced neutrophil inflammatory factors IL-2,IL-6,IL-10 and TNF-α.The morphological characteristics of exosomes were observed by electron microscope.Mir-15a-5p was significantly overexpressed in neutrophil exosomes induced by lipopolysaccharide;Lipopolysaccharide-induced neutrophil exosomes and mir-15a-5p simulants can promote raw267.4 macrophage apoptosis and inhibit its cell viability;Targetscan database predicted that mir-15a-5p and BCL2L2 had binding sites.Double-luciferase experiment verified that mir-15a-5p and BCL2L2 bound through binding sites;Mir-15a-5p mimic was transfected into raw267.4 macrophages which inhibit the expression of BCL2L2 mRNA and protein.Conclusion:inflammatory neutrophils may promote raw267.4 by secreting exosomes containing mir-15a-5p and inhibiting BCL2L2 by targeting macrophage apoptosis.This may provide a theoretical basis for further understanding the molecular mechanism of inflammatory regulation of neutrophils and macrophages.展开更多
基金National Natural Science Foundation of China (No.82060357)Hainan Natural Science Foundation (No.2019RC370,820RC757)Research Project of Hainan Provincial Health Commission (No.20A200021)。
文摘Objective:To explore the possible mechanism of neutrophil exosomes regulating macrophage apoptosis.Methods:neutrophils were induced by lipopolysaccharide,inflammatory factors were detected by ELISA,the morphology of exosomes was identified by electron microscope,and the expression of mir-15a-5p in exosomes was detected by RT-PCR;Raw267.4 macrophages was treated with neutrophil exosomes and mir-15a-5p mimic respectively,CCK8 to detect cell viability,flow cytometry to detect apoptosis;The binding sites of mir-15a-5p and BCL2L2 were predicted and verified by double luciferase experiment;RT-PCR and Western blot verified that mir-15a-5p regulate the expression of BCL2L2.Results:lipopolysaccharide induced neutrophil inflammatory factors IL-2,IL-6,IL-10 and TNF-α.The morphological characteristics of exosomes were observed by electron microscope.Mir-15a-5p was significantly overexpressed in neutrophil exosomes induced by lipopolysaccharide;Lipopolysaccharide-induced neutrophil exosomes and mir-15a-5p simulants can promote raw267.4 macrophage apoptosis and inhibit its cell viability;Targetscan database predicted that mir-15a-5p and BCL2L2 had binding sites.Double-luciferase experiment verified that mir-15a-5p and BCL2L2 bound through binding sites;Mir-15a-5p mimic was transfected into raw267.4 macrophages which inhibit the expression of BCL2L2 mRNA and protein.Conclusion:inflammatory neutrophils may promote raw267.4 by secreting exosomes containing mir-15a-5p and inhibiting BCL2L2 by targeting macrophage apoptosis.This may provide a theoretical basis for further understanding the molecular mechanism of inflammatory regulation of neutrophils and macrophages.
