Effect of miR-145-5p on macrophage polarization via TGF-β1/Smads pathway
Objective To investigate the impact of miR-145-5p on the polarization of macrophages and related mechanism.Methods Raw264.7 macrophages were stimulated with porphyromonasgingivalis-lipopolysaccharide (Pg-LPS) to induce a periodontitis model (model group).The effect of miR-145-5p on macrophage polarization was evaluated by flow cytometry;the expression levels of miR-145-5p,Arg-1,iNOS,IL-6,IL-10,TGF-β1 and Smad3 mRNA were detected by qRT-PCR;the expression of TGF-β1/Smads pathway-related proteins TGF-β1 and Smad3,and the marker proteins of the macrophages iNOS and Arg-1 were detected by Western blot.Results Under light microscopy,the model group showed an increased proportion of polygonal M1-type macrophages.In the model group transfected with miR-145-5p mimics (transfected model group),the proportion of polygonal M1-type macrophages was significantly reduced.Compared to the negative control (NC) group and non-transfected model group the expression level of miR-145-5p in the transfected model group was significantly increased (both P<0.05);the mRNA expression levels of Arg-1 and IL-6 were elevated (both P<0.05),and the mRNA levels of iNOS,IL-10,Smad3,and TGF-β1 were decreased (all P<0.05);the protein expression levels of Arg-1 were increased,and the protein levels of iNOS,TGF-β1 and Smad3 were decreased (all P<0.05).The proportions of M1-type macrophages in the transfected model group was reduced than the NC group (P<0.05),while the proportions of M2-type macrophages was increased than the NC group (P<0.05).Conclusion MiR-145-5p may inhibit mouse macrophages M1 polarization and promote M2 polarization by suppressing the Smad3 protein in the TGF-β1/Smads pathway to inhibit inflammation,which indicating that it may be a potential therapeutic agent for periodontitis treatment.
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