Effect and Mechanism of Zhuanggu Jianxi Formula on Macrophage Polarization
Objective:To investigate the effect and mechanism of Zhuanggu Jianxi Formula(ZGJXF)on the polarization of macro-phages into classical activation(M1)and alternative activation(M2)types.Methods:Phorbol 12-myristate 13-acetate was used to induce differentiation of human monocytic leukemia cells into non-activated macrophages(M0).Lipopolysaccharide was used to stimulate M0 to replicate the M1 polarization model.Cells were divided into a blank group(M0+blank serum),a model group(M1+blank serum),an inhibitor group(M1+NF-κB inhibitor+blank serum),and a ZGJXF group(M1+medicated serum).After 24 hours of intervention,various indicators were measured,including M1 markers inducible nitric oxide synthase(iNOS),major histo-compatibility complex Ⅱ(MHC-Ⅱ),monocyte chemotactic protein 1(MCP-1),M2 markers CD206,CD163,CD209 mRNA expres-sion,pro-inflammatory factors interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),anti-inflammatory factors IL-4,IL-10 levels and mRNA expression,and key proteins in the NF-κB signaling pathway,such as nuclear NF-KBp65,NF-KBp50,and NF-κB inhib-itor α(IκBα).Results:In the model group,M1 markers,pro-inflammatory factors,and NF-KBp65,NF-KBp50 proteins were signifi-cantly higher than those in the blank group(P＜0.05),while CD206,IL-4 mRNA,IKBα protein,and IL-4 levels were lower than those in the blank group(P＜0.05).In the inhibitor and ZGJXF groups,M1 markers,pro-inflammatory factors,and NF-KBp65,NF-KBp50 proteins were lower than those in the model group(P＜0.05),while M2 markers,anti-inflammatory factors,and IκBα pro-tein were higher than those in the model group(P＜0.05).The ZGJXF group had lower levels of MHC-Ⅱ,MCP-1 mRNA,and pro-inflammatory factors than the inhibitor group(P＜0.05),and higher levels of CD206,CD209,IL-10 mRNA,IL-4,and IL-10(P＜0.05).Conclusion:ZGJXF can inhibit lipopolysaccharide-induced M1 polarization of macrophages and promote M2 polarization.The mechanism may be related to the reduction of the activity of the NF-κB signaling pathway in cells.
Zhuanggu Jianxi FormulaDrug-containing serumKnee osteoarthritisMacrophage polarizationM1/M2 type macro-phageSynovitisTHP-1-derived macrophages
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