〔Abstract〕 Objective To study the effect of paeoniflorin-6-O′-benzenesulfonate(CP-25) on the expression of uptake transporter in rat fibroblast-like synoviocytes(Rat-FLS). Methods Rat-FLS were isolated and culturedin vitroand the cell state was observed by an inverted microscope. Flow cytometry and CCK-8 were used to detect the purity and growth curve of FLS, respectively. qRT-PCR was used to detect the effects of CP-25 on mRNA levels of transporters in Rat-FLS. Western blot was chosen to study the effect of CP-25 on the expression of transporters in Rat-FLS. Results Rat-FLS isolated and cultured from synovial tissue of rats showed active proliferation and favorable growth. The results of flow cytometry showed that Rat-FLS purity was over 96%. CCK-8 showed that Rat-FLS had better proliferation activity for 1-7 days. The results of qRT-PCR showed that 10 μmol/L CP-25 up-regulated organic anion transporter(OAT) 1/3, organic cation transporter(OCT) 1/3, glucose transporter(GLUT) 4 and oligopeptide transporter(PEPT) 1 mRNA levels, while CP-25 down-regulated GLUT1 and monocarboxylate transporter(MCT) 1/2 mRNA levels. Western blot showed that the expression of transporters OAT1/3, OCT1/3, GLUT4, and PEPT1 were up-regulated by CP-25. At the same time, the expression of transporters GLUT1 and MCT1/2 were down-regulated by CP-25. Conclusion CP-25 can regulate the uptake transporters to varying degrees, which provides a reference for the study of the interaction between CP-25 and other drugs.