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Objective The aim of the present study was to investigate the effect of paeoniflorin (Pae) on the recombinant human interleukin-1β (rhIL-1 β)-stimulated human peripheral blood lymphocytes (PBLC) in vitro.Methods PBLCs were collected by the Ficoll density gradient centrifugation, co-cultured with rhIL-1β (0.01, 0.1, 1, 10, 100 μg/L) for 24 h or rhIL-1β (1 μg/L) for different time periods (3, 6, 12, 24, 48 h).The proliferation response was determined by CCK-8 assay.The production of IL-17 and IL-10 were measured by ELISA.Peripheral Th1 7 and Treg frequencies were analyzed by flow cytometry (FCM).Results These results showed that the proliferative function and the activity of IL-17 produced by PBLC were promoted significantly by rhIL-1β, the levels of IL-10 was decreased on the contrary.The FCM analysis showed that rhIL-1β significantly increased the frequency of Th1 7 and decreased the frequency of Treg.However, administration of Pae (10-8, 10-7, 10-6, 10-5 mol/L) significantly suppressed proliferation responses of rhIL-1β-induced PBLC, regulated the secretion function of IL-17 and IL-10.Further study indicated Pae (10-7, 10-6, 10-5 mol/L) recovered the frequencies of Th17 and Treg in human PBLC with rhIL-1β-stimulated.Conclusions These results suggested that Pae might exert anti-inflammatory and immunomodulatory effects through recovering the imbalance of Th17/Treg.