The Inhibition Effect And Mechanism Of Chlorogenic Acid On M2 Polarization In Hepatocellular Carcinoma-associated Macrophages Through JAK/STAT6 Signaling Pathway

ObjectiveThe JAK / STAT6 signaling pathway was activated by Interleukin 4(IL-4)and interrupted by chlorogenic acid(CGA).By analyzing the differences of related molecular proteins and gene expression levels,to investigate the inhibition effect and mechanism of Chlorogenic Acid on M2 polarization in hepatocellular carcinoma-associated macrophages.MethodsWe divided the cells of RAW264.7 and Ana-1 all into six groups: model group,normal control group,high dose CGA group,CGA middle dose group,low dose group CGA group and Oxaliplatin group.The cells in the other groups were treated with 50 ng / ml IL-4 and IL-4 for 24 hours,and corresponding concentration were added to the cells for 12 hours.Then cells and supernatant were collected.Before and after the IL-4 stimulated cells,the m RNA expression of KLF4,SOCS1,PPAR?,PPAR? were detected by RT-PCR at the level of gene transcription,the protein expression of KLF4,SOCS1,PPAR? and PPAR? were detected by Western Blot,the expression levels of TGF—? was detected by ELISA.ResultsContracted with the nomal group,the m RNA expression levels of SOCS1,KLF4,PPAR?,PPAR? of model group were significantly increased in RAW264.7 cells and Ana-1 cells,the protein levels of IL-4R??STAT6 do not change,the SOCS1,KLF4,PPAR? and PPAR? were increased,the level of TGF — ? was increased.The differences have statistical significance(P <0.05).Contracted with the model group,the m RNA expression level of SOCS1,KLF4,PPAR? and PPAR?,the protein expression of p STAT6,KLF4,SOCS1,PPAR? and PPAR? were decreased in a dose-dependent manner,and the expression of TGF—?was also considerably decreased,The differences have statistical significance(P<0.05).OXA had a certain degree of inhibition of PPAR? and PPAR? at protein level and gene level,but the inhibition effect was not as good as CGA.Conclusions1.It was confirmed that CGA could inhibit the polarization of M2 type of tumor-associated macrophage.2.JAK / STAT6 signaling pathway is one of the potential molecular mechanisms of CGA inhibition of M2 polarization in hepatocellular-associated macrophages.