〔Abstract〕 Objective To investigate effect 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 into control group, tumor necrosis factor-α(TNF-α) 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 detect the expression of miR-20a-5p in HFLS-RA cells; enzyme-linked immunosorbent assay(ELISA) was applied to detect the levels of interleukin-1β(IL-1β) and IL-6 in the supernatant 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 HFLS-RA cell migration; Western blot was applied to detect the expression of TIMP2, 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 IL-1β and IL-6, OD450value, EdU positive cell rate, scratch healing rate, and the expression of CyclinD1 and MMP-9 proteins in the TNF-α group increased, the expression ofTIMP2 protein decreased(P<0. 05); compared with the TNF-α group, the expression of miR-20a-5p, the levelsof IL-1β and IL-6, OD450value, EdU positive cell rate, scratch healing rate, and CyclinD1 and MMP-9 proteins expression decreased, the expression of TIMP2 protein increased in salidroside group(P<0. 05); compared withthe TNF-α group and inhibitor NC group, the expression of miR-20a-5p, the levels of IL-1β and IL-6, OD450value, EdU positive cell rate, scratch healing rate, and the expression of CyclinD1 and MMP-9 proteins in the miR-20a-5p inhibitor group decreased, the expression of TIMP2 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-1β and IL-6,OD450value, EdU positive cell rate, scratch healing rate, and the expression of CyclinD1 and MMP-9 proteins in the 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 HFLS-RA cell proliferation, migration and inflammatory response by regulating miR-20a-5p/TIMP2.