Hydrogen therapy promotes macrophage polarization to the M2 subtype by inhibiting the NF-κB signaling pathway
Objective To investigate the role of hydrogen therapy in reducing radiation-induced lung injury and the spe-cific mechanism.Methods Forty C57BL/6 mice were randomly divided into four groups:normal control group,model group,hydrogen therapy group I,and hydrogen therapy group Ⅱ.A mouse model of radiation-induced lung injury was estab-lished.The pathological changes in the lung tissue of the mice were examined with HE staining.Immunofluorescence stain-ing was used to detect the expression of surface markers of Ml and M2 macrophages to observe macrophage polarization.The expression of interleukin(IL)-6,tumor necrosis factor-α(TNF-α),and IL-10 in the lung tissue was measured by immun-ohistochemistry.The expression of nuclear factor-kappa B(NF-κB)p65 and phosphorylated NF-κB(P-NF-κB)p65 was measured by Western blot.Results HE staining showed that compared with the control group,the model group exhibited alveolar septal swelling and thickening,vascular dilatation and congestion,and inflammatory cell infiltration in the lung tis-sue;the hydrogen groups had significantly reduced pathological damage and inflammatory response than the model group,with more improvements in hydrogen group Ⅱ than in hydrogen group I.Immunohistochemical results showed that com-pared with those in the control group,the levels of the inflammatory cytokines IL-6 and TNF-α were significantly increased in the model group;the hydrogen groups showed significantly decreased IL-6 and TNF-α levels and a significantly increased level of the anti-inflammatory factor IL-10 than the model group,which were more marked in hydrogen group Ⅱ than in hy-drogen group I.Immunofluorescence results showed that compared with the control group,the expression of the surface marker of M1 macrophages in the model group was significantly upregulated;the hydrogen groups showed significantly downregulated M1 marker and significantly upregulated M2 marker,and hydrogen group Ⅱ showed significantly increased M2 marker compared with hydrogen group Ⅰ.Western blot results showed that compared with that in the control group,the ratio of P-NF-κB p65/NF-KB p65 in the model group was significantly increased;the P-NF-κB p65/NF-KB p65 ratio was sig-nificantly reduced in the hydrogen groups than in the model group,and was significantly lower in hydrogen group Ⅱ than in hydrogen group Ⅰ.Conclusion Hydrogen inhalation therapy may reduce the inflammatory response of radiation-induced lung injury by inhibiting the NF-κB signaling pathway to promote the polarization of the macrophage Ml subtype to the M2 subtype.
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