Matrine is a kind of alkaloid found in certain Sophora plants, which has been extensively used in China for the treatment of viral hepatitis, cancer, cardiac diseases and skin diseases (such as atopic dermatitis and eczema). It also has been confirmed that substance P (SP) and its receptor (neurokinin-1 receptor, NK-1R) are involved in the pathogenesis of inflammatory skin disorders. So the present study was designed to investigate the effect of matrine on the expression of NK-1R and cytokines production induced by SP in HaCaT cells (a human epidermal keratinocyte cell line) and dermal fibroblasts. In addition, cell viability was also evaluated. The results showed that matrine inhibited the expression of NK-1R in HaCaT cells and fibroblasts. SP induced the production of interleukin (IL)-1beta, IL-8, interferon (IFN)-gamma, and monocyte chemotactic protein (MCP)-1 in both cell types. Matrine 5-100 microg/mL had little effect on cell viability. It inhibited SP-induced IL-1beta, IL-8 and MCP-1 production in HaCaT cells and fibroblasts, while it increased the production of IFN-gamma in HaCaT cells. Both SP and matrine had no effect on the secretion of IL-6. These findings suggest that matrine may have potential treatment function on SP related cutaneous inflammation by inhibition of the expression of substance P receptor and regulation of the production of inflammatory cytokines.