Inhibition of lipopolysaccharide-induced RAW 264.7 cell inflammation by human bone marrow mesenchymal stem cells via exosomes
Objective:To investigate the anti-inflammatory effect of bone marrow mesenchymal stem cell exosomes(MSC-Exo)in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells and its possible mechanisms.Methods:Mesenchymal stem cells(MSCs)were extracted from mouse bone marrow and cultured with the primary culture method.Mesenchymal stem cell conditioned medium(MSC-CM)and exosomes were extracted to verify the inhibitory effect of MSC-CM and MSC-Exo on LPS-induced inflammation in RAW 264.7 cells.RAW 264.7 cells were stimulated by LPS for 4-6 h,and cell proliferation activity was detected by cell counting and MTT assay.The exosomes in MSC-CM were detected by Western blot.The morphology and particle size of the exosomes were observed with the particle size analyzer.The morphological changes of cells were observed by inverted fluorescence microscope.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect mRNA transcription levels of IL-6 and Arg-1 in RAW 264.7 cells induced by LPS.Enzyme-linked immunosorbent assay(ELISA)was used to evaluate the secretion of IL-6 and IL-10 in LPS-induced RAW 264.7 cells.Results:Compared with LPS group,MSC-Exo inhibited cell differentiation and significantly decreased the secretion of the cytokine IL-6 in LPS-induced cells after 30 h(P<0.001).MSC-Exo inhibited IL-6 mRNA transcription,and promoted Arg-1 mRNA transcription,and increased IL-10 expression.Conclusion:This study preliminarily demonstrated that bone marrow mesenchymal stem cell exosomes can inhibit lipopolysaccharide-induced RAW 264.7 cell inflammation.
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