Object To study the effect of puerarin on the expression of inflammatory factors and miR-155-3p in human umbilical vein endothelial cells (HUVEC) induced by visfatin. Methods The HUVEC cell injury model was established with visfatin. Cell proliferation was measured by MTT assay. Cell apoptosis was detected by flow cytometry. The level of CRP and NF-κB was detected by ELISA, and the expression of miR-155-3p was detected by RT-PCR. The expression of myeloid differentiation factor 88 (MyD88) was identified by western blotting. Results Visfatin induced cell proliferation and inhibited apoptosis in HUVEC, meanwhile the expressions of both CRP and NF-κB were significantly increased, compared with that of the control group (P<0.01). Puerarin at moderate and high concentrations obviously reduced the HUVEC injury induced by visfatin, mainly through down-regulating the expression of CRP and NF-κB, as well as up-regulating the level of miR-155-3p in the HUVEC. MiR-155-3p mimic markedly decreased the level of MyD88, CRP and NF-κB in the HUVEC induced by visfatin (P<0.05). Conclusions Pueprarin obviously alleviates HUVEC injury induced by visfatin, probably related to down-regulating the level of MyD88, CRP, NF-κB, and up-regulating the expression of miR-155-3p in HUVEC.