This study provides in vitro and in vivo evaluation of rat serum metabolites of the Pueraria lobata (SMP) on peripheral nerve regeneration. In the in vitro study, we found that the SMP caused a marked enhancement of the nerve growth factor (NGF)-mediated neurite outgrowth and the expression of synapsin I from PC12 cells. In the in vivo study, silicone rubber chambers filled with the SMP were used to bridge a 10-mm sciatic nerve defect in rats. At the conclusion of 8 weeks, animals from the groups treated with the SMP had a relatively more mature structure with larger mean values of myelinated axon number, endoneurial area, and total nerve area when compared with those in the controls receiving the saline only. These results suggest that the serum metabolites of Pueraria lobata can be a potential nerve growth-promoting factor.