The imbalance between effector CD4+ T helper?17 (Th17) and regulatory CD4+ T cells (Treg) cells and their associated cytokines, have been associated with the pathogenesis of inflammatory bowel disease (IBD). Total glycosides of paeony (TGP) is an alternative immunomodulatory agent that is widely used for the treatment of autoimmune diseases. The present study aimed to evaluate the modulatory effect of TGP in a rat model of colitis induced by 2,4,6?trinitrobenzene sulfonic acid (TNBS). TGP was administered intragastrically 24?h after the TNBS intrarectal instillation for 7?days. TGP treatment ameliorated the clinical status and reversed the histopathologic severity of acute TNBS colitis. Furthermore, TGP inhibited the levels of Th17?associated cytokines interleukin (IL)?17, IL?6, tumor necrosis factor?α, whereas the expression levels of Treg?associated cytokines IL?10, transforming growth factor?β in the plasma, colon, spleen and mesenteric lymph nodes (MLN). Additionally, TGP reduced the percentage of Th17 cells; however, the proportion of Treg cells in the spleen and MLN was increased. The present study also observed a suppression of Th17?associated transcription factor, termed retinoid?related orphan receptor?γt (ROR?γt). However, expression of the Treg?associated transcription factor forkhead boxp3 was increased in the TGP treatment group. Therefore, the present findings suggest that TGP has a regulatory role in modulating the balance of Th17 and Treg cells to ameliorate the TNBS?induced colitis and support the strategy of using TGP to treat IBD.