Effects Of Hepatocyte Growth Factor On The Polarization Of Ml And M2 Subtypes In Mouse Macrophages

Objective In vitro conditions,to investigate the effect of HGF on macrophage polarization by analyzing the effects of different concentrations of HGF on macrophage M1 markers arginase II(Arg-II),inducible nitric oxide(iNOS)and interleukin-6(IL-6)and M2 marker arginase I(Arg-I)and interleukin-10(IL-10).Methods Mouse macrophage cell line RAW264.7 cells were cultured in vitro and randomly divided into three groups: M?(RAW264.7 cells),M1 group(M1 macrophages)and M2 group(M2 macrophages).M? group was used as a blank control group,M1 group was induced by 20 ng/ml interferon-?(IFN-?)combined with 500ng/ml bacterial lipopolysaccharide(LPS),M2 group induced polarization at 20 ng/ml interleukin-4(IL-4).Among them,M1 and M2 groups were treated with 0.1,1.0 and 10 ng/ml HGF after induced polarization for 12 hours,followed with coculture 24 hours,collected the cells and their supernatants.Inverted microscope was used to observe the cell morphology of M?,M1 and M2 groups;Western blot was used to detect the protein expression of Arg II,a marker of M1 type,and Arg I,a marker of M2 type macrophage;The immunoflurescence assay was used to detect the M1 marker iNOS and the M2 marker Arg I;ELISA method was used to detect the concentration of IL-6,a marker of M2,and IL-10,a marker of M2 in the cell supernatant.Results 1.M? macrophages were round,there were many antennae around M1 macrophages,M2 macrophages were "omelet-like" shape;2.HGF had no significant proliferative effect on M? group,M1 group and M2 group;3.Compared with M? group,the expression levels of Arg I,IL-10 in Arg II,iNOS,IL-6 and M2 groups were significantly increased(P <0.05).4.Compared with M1 group,the expression of Arg II,iNOS and IL-6 in HGF intervention group was significantly lower than that in M1 group after HGF intervention(P<0.05);Compared with M2 group,Arg I and IL-10 significantly increased(P<0.05),and 1.0,10 ng /ml HGF treatment group showed a dose-dependent manner(P<0.01).Conclusion 1.In vitro,IFN-? combined with LPS induced the polarization of RAW264.7 macrophages to M1,and IL-4 induced the polarization of RAW264.7 macrophages to M2;2.In vitro,different concentrations of HGF can inhibit the polarization of RAW264.7 macrophages to M1 and promote the polarization of RAW264.7 macrophages to M2.