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Present study was carried out to investigate the effect of herbal complex extract including Houttuynia cordata Thunb on hair growth and gene expression in C57BL/6 mice. Twenty mice were divided into 4 experimental groups including normal (control), 50% herbal complex extract (S-1), 100% herbal complex extract (S-2) and 5% minoxidil-treated group. The test materials were daily applied with 200 μl per mouse on shaved dorsal skin for 3 weeks. The hair growth was monitored by photograph at 7, 14 and 21 days after topical application. Then the changes of hair density and hair thickness in the hair-removed area were evaluated by phototrichogram using folliscope. Also the expression level of growth factors related to hair growth was measured by real time polymerase chain reaction. Application of S-2 or minoxidil-treated group stimulated the hair growth compared to vehicle treatment. Therefore hair density of S-2 or minoxidil-treated group application was increased about 2.5 and 3.1 fold more than in vehicle application on 21 day, respectively. And hair weight of both S-2 and minoxidil-treated group were increased about 8.3 and 10.0 fold more than in vehicle spreading on 21 day, respectively. Futhermore the gene expression of insulin-like growth factor (IGF)-1 and vascular endothelial growth factor (VEGF) were significantly up-regulated in S-2 and minoxidil-spreaded group compared to vehicle-applied group. However the gene expression of transforming growth factor (TGF)-β1 was not significantly down-regulated in S-2 and minoxidil-spreaded group compared to vehicle-applied group. These data suggest that S-2 has potent stimulating activity on hair growth in C57BL/6 mice and potential usefulness as ingredients of hair tonic and hair restore.