P#45  Isolation and identification of barley derived hair-growth stimulant

Ayako Kamimura, Tomoya Takahashi. Kyowa Hakko Kogyo, Tsukuba, Japan

With the aim of identifying natural products which possess hair-growing activity, we examined more than 1,000 plant extracts with respect to their growth-promoting effects on murine hair epithelial cells. We discovered intensive growth-promoting activity, about 140% relative to controls, in barley extract. Our strategy for identifying active compounds in barley extract involved subjecting it to column chromatography and subjecting each fractions to in vivo hair-growing assay using C3H mice. The 60% (v/v) aqueous methanol eluted fraction from the HP-20 column and the 75% (v/v) aqueous methanol eluted fraction from the subsequent LH-20 column showed high hair-growing activity in in vivo C3H mouse assay system. We isolated two major substances from the LH-20 active fraction using preparative HPLC. By means of mass spectrometry, 1H-NMR, and 13C-NMR analyses, one substance was revealed to be procyanidin B-3 and the other substance was identified as (+)-catechin. Purified procyanidin B-3 showed high hair-growing activity (30 mM=302%) in the form of in vitro hair epithelial cell growth-promoting activity and in vivo anagen-inducing activity; however, (+)-catechin, that is monomeric unit of procyanidin B-3, showed no hair-growing activity. For the purpose of examining the hair-growing mechanisms of procyanidin B-3, we examined its relationship to the TGF-b signal pathway, which is known to be a regulator of catagen induction. Addition of TGF-b1 to hair epithelial cell cultures dose-dependently decreased the cell growth, and addition of procyanidin B-3 to the culture neutralized the growth-inhibiting effect of TGF-b1. From these results, it is concluded that procyanidin B-3 can directly promote hair epithelial cell growth in vitro, has the potential to counteract the growth-inhibiting effect caused by TGF-b1 in vitro, and has potential to stimulate anagen induction in vivo