Effects of salidroside on proliferation and migration of fibroblastoid synovial cells in rheumatoid arthritis by regulating miR⁃20a⁃5p/TIMP2 axis / 安徽医科大学学报

ABSTRACT

Objective @#To investigate ellect of salidroside on the function and activation of rheumatoid arthritis fibroblast-like synoviocyte(HFLS-RA)by regulating the miR-20a-5p/tissue inhibitor of metalloproteinase-2(TIMP2) axis.@*Methods@#HFlS-RA cells were used as the research object. HFlS-RA cells were separated intocontrol group, tumor necrosis factor-a (TNF-a) group, salidroside group, inhibitor NC group, miR-20a-5p inhibitor group, salidroside + mimic NC group, and salidroside + miR-20a-5p mimic group. qRT-PCR was applied to deteet the expression of miR-20a-5p in HFIS-RA cells ; enzyme-linked immunosorbent assay( ELISA) was applied todetect the levels of interleukin-18 ( lL-1β) and IL-6 in the supermatant of HFLS-RA cells: cell counting kit-8(CCK-8) method and 5-ethynyl-2 '-deoxyuridine ( EdU) staining were applied to detect HFLS-RA cell proliferation ; scratch experiment was applied to detect HilS-RA cell migration; Western blot was applied to detect the ex.pression of 'TlMP2, CyclinD1, and matrix metalloproteinase ( MMP ) -9 proteins in HFLS-RA cells; double lucifer.ase was applied to verify the relationship between miR-20a-5p and TIMP2. @*Results@#Compared with the control group, the expression of miR-20a-5p, the levels of lL-1β and IL-6, 0Dso value, EdU positive cell rate, scratchhealing rate, and the expression of CyclinDl and MMP-9 proteins in the TNF-α group increased, the expression of TlMP2 protein decreased ( P <0. 05 ) ; compared with the TNF-α group, the expression of miR-20a-5p, the levelsof lL.-1β and IL-6, OD450 value, EdU positive cell rate, scratch healing rate, and CyclinD1 and MMP-9 proteinsexpression decreased, the expression of TlMP2 protein increased in salidroside group ( P <0. 05 ); compared withthe 'T'NF -a group and inhibitor NC group, the expression of miR-20a-5p, the levels of IL-1 β and IL.-6, OD450 val-ue, EdU positive cell rate, seratch healing rate, and the expression of CyclinDl and MMP-9 proteins in the miR.20a-5p inhibitor group decreased, the expression of TlMP2 protein increased ( P <0. 05 ); compared with the sali.droside group and the salidroside + mimic NC group, the expression of miR-20a-5p, the levels of IL-1 β and IL-6 ,OD.so value, EdU positive cell rate, scratch healing rate, and the expression of CyelinD1 and MMP-9 proteins inthe salidroside + miR-20a-5p mimic group increased, the expression of TIMP2 protein decreased ( P < 0. 05 )There was a targeted regulatory relationship between miR-20a-5p and TIMP2. @*Conclusion@#Salidroside may inhibit TNF-α-induced HFS-RA cell proliferation , migration and infammatory response by regulating miR-20a-5p/TIMP2.