Over-expression of MMP-7 regulates the secretion of Gal-3 in macrophages and influences their polarization and immune responses
To explore the effect of matrix metalloproteinase 7(MMP-7)over-expression on macrophage polarization,galectin 3(Gal-3)and inflammatory factors secretions,THP-1 cells were induced to macrophages and transfected with MMP-7 over-expressing plasmid or empty vector.The cells were divided into control group,MMP-7 over-expressing group,MMP-7 NC group,M1-polarized group,and M2-polarized group.mRNA expression levels of Gal-3 and MMP-7 were quantified by qPCR.The percentages of F4/80+,CD86+,and CD206+macrophages were detected by flow cytometry.The expressions of inflam-matory cytokines IL-1α,IL-6,and TGF-β were detected by ELISA.After Gal-3 inhibitor treatment on M1-induced and M2-induced macrophages,the mRNA expression of Gal-3 in above macrophages was detected by qPCR,and the levels of IL-1α,IL-6,and TGF-β in the supernatant of cell cultures were detected by ELISA.The results showed that compared to the control group,the MMP-7 over-expressing group and the M2-polarized group showed significant increase in the expressions of Gal-3 and MMP-7 mRNA,in the percentage of(F4/80+)/CD206+macrophages(all with P＜0.01),and in the concentrations of TGF-β(both P＜0.01),while IL-1α and IL-6 levels showed significant decrease(both P＜0.01).In the M1-polarized group,the percentage of(F4/80+)/CD86+macrophages increased significantly(P＜0.01).The concentration of TGF-β decreased,while the concentrations of IL-1α and IL-6 increased significantly(all with P＜0.01).Compared to that of the M1-polarized group,the rela-tive expression of Gal-3 mRNA in the M1+Gal-3 inhibitor group was significantly decreased(P＜0.01),the concentration of TGF-βsignificantly increased(P＜0.01),while the IL-1α and IL-6 levels significantly decreased(both P＜0.01).Compared to those of the M2-polarized group,the relative expression of Gal-3 mRNA in M2+Gal-3 inhibitor group was significantly decreased(P＜0.05),along with decreased IL-1α and IL-6 and increased TGF-β concentrations(all with P＜0.05).This study demonstrates that MMP-7 can activate Gal-3 expression to promote M2-type macrophage polarization and present anti-inflammatory activity.
NETL
NSTL
万方数据
