Paeoniflorin inhibits the inflammatory response of bronchial epithelial cells by mediating the polarization of THP-1-derived macrophages
To investigate the effect and mechanism of paeoniflorin(PF)on human bronchial epithelial cells(BEAS-2B),THP-1 cells were induced to differentiate into macrophages by phorbol 12-myristate 13-acetate,and the toxicity of PF to THP-1 macrophages was detected by CCK-8 assay.THP-1 macrophages were cultured with 1 μg/mL LPS to induce inflammation,and treated with 1,10,and 30 μmol/L PF for 24,48,and 72 h,respectively.Cell viability was detected by CCK-8 assay,and the optimal concentration and treating time of PF were determined.BEAS-2B cells were inoculated in the upper chamber of Transwell and THP-1 macrophages were inoculated in the lower chamber of Transwell.THP-1 macrophages were divided into 4 groups: control group,LPS group,PF group,and LPS+PF group.The survival rate of BEAS-2B cells was detected by CCK-8 assay,the apoptosis rate of BEAS-2B cells was detected by FACS,the levels of inflammatory cytokines IFN-γ,IL-4,IL-17C,and IL-10 were detected by ELISA,and the expression levels of CD63,CD9,and apoptosis-linked gene 2-interaction protein X(Alix)were detected by Western blotting.The expressions of M1 and M2 markers CD80 and CD206 in THP-1 macro-phages were detected by FACS.The results showed that the optimal concentration and treating time of PF were 10 μmol/L and 48 h.Compared to those in the control group,the cell survival rate,IL-4 and IL-10 levels and the proportion of M2-type macrophages in the LPS group were significantly decreased(P＜0.01).On the other hand,the apoptosis rate,IFN-γ and IL-17C levels,CD63,CD9,Alix protein expression levels and the proportion of M1-type macrophages were significantly increased(P＜0.01).Compared to those in the LPS group,the cell survival rate,IL-4 and IL-10 levels and the proportion of M2-type macrophages in the LPS+PF group were significantly increased(P＜0.01),while the apoptosis rate,IFN-γ and IL-17C levels,CD63,CD9,Alix protein expression levels and the proportion of M1-type macrophages were significantly decreased(P＜0.01).These results suggest that PF can improve the survival rate of BEAS-2B cells,reduce apoptosis,and inhibit exosome secretion and inflammatory response.The underlying mechanism may be related to PF induced M2-type polarization of macrophages.
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