Expression changes of LYVE1+ macrophages in synovial tissues of joints in patients with RA and their inhibitory effects on migration,invasion,and FMT
Objective To observe the expression changes of lymphatic vessel endothelial receptor-1(LYVE1)+ mac-rophages in the synovial tissues of patients with rheumatoid arthritis(RA)and their inhibitory effects on migration,inva-sion,and fibroblast-to-myofibroblast transformation(FMT).Methods Immunofluorescence staining was used to qualita-tively and quantitatively detect LYVE1 and CD68 in synovial tissues of 45 RA patients and 45 osteoarthritis(OA)pa-tients.Human monocytic leukemia THP-1 cells in the logarithmic growth phase were treated with LYVE1 overexpression lentivirus in the culture medium,and LYVE1-expressing THP-1 cells were obtained after 48 h of cultivation.These cells were induced with 100 ng/mL phorbol 12-myristate 13-acetate(PMA)for 48 h to obtain LYVE1+ macrophages.Another portion of THP-1 cells were induced by only 100 ng/mL PMA for 48 h to obtain LYVE1-macrophages.Human rheumatoid arthritis fibroblasts MH7A in the logarithmic growth phase were divided into the LYVE1+ macrophage group and LYVE1-macrophage group,which were added with LYVE1+ macrophages and LYVE1-macrophages,separately.In addition,the cells containing only medium were set as the blank control group.Scratch experiment was performed to observe the migra-tion ability of cells in each group.MH7A cells were divided into groups A,B,and C,which were added with LYVE1+ macrophages,LYVE1-macrophages,and only culture medium,respectively.Transwell invasion experiment was conduct-ed to observe the invasion ability of cells in each group.MH7A cells were divided into the group one,group two,and the blank control group,which were added with LYVE1+ macrophages,LYVE1-macrophages,and only culture medium,re-spectively.Real-time quantitative PCR was used to detect the mRNA expression of FMT-related genes(COL1A1,fibro-nectin,α-SMA)in MH7A cells after 48 h of culture.Results The expression locations of LYVE1 and CD68 in the syno-vial tissues of RA and OA patients overlapped.The relative expression levels of LYVE1 in the synovial tissues of RA and OA patients were 0.319±0.033 and 1.000±0.159,respectively,with statistically significant difference between these two groups(both P<0.05).Compared with the LYVE1-macrophage group and the blank control group,the LYVE1+ macro-phage group had lower cell scratch healing at 24 and 48 h of cultivation(all P<0.05).Compared with groups B and C,the number of transmembrane cells in the group A was smaller at 24 h of cultivation(P<0.05).Compared with groups two and the blank control group,the relative expression levels of COL1A1 mRNA,fibronectin mRNA,and α-SMA mRNA in the group one were lower at 48 h of cultivation(all P<0.05).Conclusion LYVE1+ macrophages are underexpressed in the synovial tissues of RA patients,and LYVE1+ macrophages can inhibit the migration,invasion,and FMT.
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