Aim To assess the effect of acetylcholine(Ach) and its antagonist methyllycaconitine(MLA) on murine preadipocytes. Methods MTT was used for the proliferation ability examination.ELISA was used for IL-6 and IL-10.Western blotting was used for NF-κB p65 protein in nucleus. Results Ach could promote preadipocytes proliferation,with 122.0%±11.4% compared to control after stimulation for 12 h,and MLA could block its effect.Ach could induce preadipocytes secret IL-6 and reach its peak at 6 h,significantly higher than that of control group(P><0.05).Ach could decrease IL-10 concentration.MLA had little effect on cytokines secretion.Ach could increase the level of NF-κB p65 in nucleus,with the highest level at 6 h.MLA couldn't block its effect. Conclusion Ach could activate preadipocytes through promoting NF-κB p65 protein translocation.MLA couldn't antagonize its action,suggesting that ɑ7 n Ach receptor,which is the major receptor through which Ach exerts its anti-inflammation effect,didn't play an important role in the process.