Treatment of Sprague-Dawley rats with
AY9944, an inhibitor of 3β-hydroxysterol-Δ(7)-reductase (Dhcr7), leads to elevated levels of
7-dehydrocholesterol (7-DHC) and reduced levels of
cholesterol in all
biological tissues, mimicking the key biochemical hallmark of
Smith-Lemli-Opitz syndrome (SLOS). Fourteen 7-DHC-derived
oxysterols previously have been identified as products of
free radical oxidation in vitro; one of these
oxysterols, 3β,5α-dihydroxycholest-7-en-6-one (DHCEO), was recently identified in Dhcr7-deficient cells and in brain tissues of Dhcr7-null mouse. We report here the isolation and characterization of three novel 7-DHC-derived
oxysterols (4α- and 4β-hydroxy-7-DHC and 24-hydroxy-7-DHC) in addition to DHCEO and
7-ketocholesterol (7-kChol) from the brain tissues of AY9944-treated rats. The identities of these five
oxysterols were elucidated by HPLC-ultraviolet (UV), HPLC-MS, and 1D- and 2D-NMR. Quantification of 4α- and 4β-hydroxy-7-DHC, DHCEO, and 7-kChol in rat brain, liver, and serum were carried out by HPLC-MS using d(7)-DHCEO as an internal standard. With the exception of 7-kChol, these
oxysterols were present only in tissues of AY9944-treated, but not control rats, and 7-kChol levels were markedly (>10-fold) higher in treated versus control rats. These findings are discussed in the context of the potential involvement of 7-DHC-derived
oxysterols in the pathogenesis of SLOS.