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OBJECTIVE To investigate the potential molecular mechanism of Paeoniflorin (Pae) in inhibiting PGE2 induced human fibroblast-like synoviocytes (hFLS) hyperplasy.METHODS HFLS were cultured in prostaglandin E2 (PGE2) with or without different concentrations of Pae.Synoviocyte proliferations were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl (MTT)assay.The expressions of EP2, Gαts, β-arrestin2, PKC in hFLS were measured by western blot or flow cytometry.The mRNA change of EP2 was detected by RT-PCR.Cyclic adenosine monophosphate (cAMP) levels in synoviocytes were assessed by radioimmunoassay (RIA).RESULTS Pae significantly inhibited hFLS proliferation induced by PGE2 via increasing cAMP levels.High levels of total EP2, β-arrestin2 and PKC expression and EP2 mRNA in hFLS induced by PGE2 were restored with different Pae administration.EP2 downregulated and β-arrestin2 upregnlated in hFLS membrane after PGE2 stimulation, however, both were improved by Pae in various degrees.CONCLUSION Our findings suggest Pae inhibit human fibroblast-like synoviocytes proliferation induced by PGE2 via arresting EP2 receptor internalization by inhibiting β-arrestin2 recruit to cell membrane.