Biomedical subjects

A C Cotillon

Publications and source records attributed to A C Cotillon.

6 recordsLinked to original sources

PubMed2000

7 alpha-hydroxy-dehydroepiandrosterone and immune response.

In human and murine lymphoid organs, circulating 3 beta-hydroxysteroids, including pregnenolone (PREG), dehydroepiandrosterone (DHEA), and epiandrosterone (EPIA), are 7 alpha-hydroxylated by a cytochrome P450 identified in the hippocampus as P4507B1. Mouse and human lymphoid organs produced different patterns of 3 beta-hydroxysteroid 7 alpha-hydroxylation with the absence of pregnenolone and epiandrosterone hydroxylation in human and mouse, respectively. Both 7 alpha-hydroxy-DHEA and 7 alpha-hydroxy-EPIA triggered a significant increase of antitetanus toxoid and anti-Bordetella pertussis toxins IgGs production in cultures of activated B + T cells derived from human tonsils, whereas both 7 alpha-hydroxy-PREG and 7 alpha-hydroxy-DHEA increased the immune response in mouse. Paracrine action of 7 alpha-hydroxysteroids resulted from their production in cells of the lymphoid organs. Comparison of P4507B1 sequences in rat, human, and two mouse species showed that one amino acid change might explain important differences in KM for 7 alpha-hydroxylation, and suggested that such differences might contribute to the extent of immune response.

Animals↗

PubMed1999

Transformation of 3-hydroxy-steroids by Fusarium moniliforme 7alpha-hydroxylase.

Transformation of physiologically important 3-hydroxy-steroids by the DHEA-induced 7alpha-hydroxylase of F. moniliforme was investigated. Whereas DHEA was almost totally 7alpha-hydroxylated, PREG, EPIA and ESTR were only partially converted into their 7alpha-hydroxylated derivatives because hydroxylation at other undetermined positions as well as reduction of ketone at C17 or C20 into hydroxyl also occurred. Cholesterol was not transformed by the enzyme. Kinetic parameters of the 7alpha-hydroxylation for these substrates were determined and confirmed that DHEA was the best substrate of the 7alpha-hydroxylase. Inhibition studies of DHEA 7alpha-hydroxylation by the other 3-hydroxy-steroids were also carried out and proved that DHEA, PREG, EPIA and ESTR shared the same active site of the enzyme. Induction effects of these steroids were compared, and DHEA appeared to be the best inducer of the 7alpha-hydroxylase of F. moniliforme.

Androsterone↗

PubMed1998

Inhibition studies of dehydroepiandrosterone 7 alpha- and 7 beta- hydroxylation in mouse liver microsomes.

Hydroxylations of dehydroepiandrosterone (DHEA) at the 7 alpha- and 7 beta- positions have been reported in numerous murine tissues and organs, including liver, and the responsible cytochrome P450 (P450) species await identification. Using thin layer chromatography and gas chromatography-mass spectrometry, we report identification of 7 alpha-hydroxy-DHEA and 7 beta-hydroxy-DHEA metabolites produced in mouse liver microsome digests and kinetic studies of their production with apparent KM values of 3.19 +/- 0.292 microM and 2.82 +/- 0.241 microM for 7 alpha- and 7 beta-hydroxylation, respectively. Investigation of P450 inhibitor and of steroid hormone effects on both 7 alpha- and 7 beta-hydroxylation of DHEA showed that, 1) different P450s were involved in 7 alpha- and 7 beta-hydroxylation of DHEA because metyrapone inhibited solely 7 alpha-hydroxylation, 2) P450 2D6, 2B1, and 2B11 were not responsible for 7 alpha- and 7 beta-hydroxylation of DHEA because respective specific inhibitors quinidine and chloramphenicol triggered no inhibition, 3) aside from P450 7b, P450 1A1, and 1A2 may be responsible for a fraction of DHEA 7 alpha- and 7 beta-hydroxylation because alpha-naphthoflavone and furafylline, which inhibit specifically P450 1A1 and 1A2, decreased the 7 alpha- and 7 beta-hydroxylation partly, 4) comparison of these findings with those obtained with brain microsomes suggested that tissue-specific P450 species are responsible for the 7 alpha- and 7 beta-hydroxylation of DHEA, 5) 7 alpha-hydroxylation of DHEA may be shared with other 3 beta-hydroxysteroids, such as 3 beta-hydroxy-5 alpha-androstan-17-one, 5-androstene-3 beta,17 beta-diol and pregnenolone, which acted in a noncompetitive manner. Taken together, these findings will be of use for identification of the P450 species responsible for 7 alpha- and 7 beta-hydroxylation of DHEA and for studies of their activities in liver.

Animals↗

PubMed1997

The inducible and cytochrome P450-containing dehydroepiandrosterone 7alpha-hydroxylating enzyme system of Fusarium moniliforme.

7Alpha-hydroxylation of DHEA by Fusarium moniliforme was investigated with regard to inducibility and characterization of the responsible enzyme system. Using GC/MS, the 7-hydroxylated metabolites of DHEA produced after biotransformation by Fusarium moniliforme mycelia were identified. The strain of Fusarium moniliforme hydroxylated DHEA predominantly at the 7alpha-position, with minor hydroxylation occurring at the 7beta-position. Constitutive 7alpha-hydroxylation activity was low, but DHEA induced the enzyme complex responsible for 7alpha-hydroxylation via an increase in protein synthesis. DHEA 7alpha-hydroxylase was found to be mainly microsomal, and the best production yields of 7alpha-hydroxy-DHEA (28.5 +/- 3.51 pmol/min/mg protein) were obtained with microsomes prepared from 18-h-induced mycelia. Kinetic parameters (KM=1.18 +/- 0.035 microM and Vmax=909 +/- 27 pmol/min/mg protein) were determined. Carbon monoxide inhibited 7alpha-hydroxylation of DHEA by microsomes of Fusarium moniliforme. Also, exposure of mycelia to DHEA increased microsomal P450 content. These results demonstrated that: (i) DHEA is 7alpha-hydroxylated by microsomes of Fusarium moniliforme; (ii) DHEA induces Fusarium moniliforme 7alpha-hydroxylase; (iii) this enzyme complex contains a cytochrome P450.

Carbon Monoxide↗

PubMed1997

Dehydroepiandrosterone 7alpha- and 7beta-hydroxylation in mouse brain microsomes. Effects of cytochrome P450 inhibitors and structure-specific inhibition by steroid hormones.

Presently, several works question the effects of dehydroepiandrosterone (DHEA) reported in vivo and designate its 7-hydroxylated metabolites as native antiglucocorticoids and potent mediators in the triggering of immune response. Among mouse tissues and organs, and second to liver, the largest production of 7alpha-and 7beta-hydroxylated derivatives of DHEA takes place in brain microsomes. To contribute to identification of cytochromes P450 (CYPs) responsible for 7alpha- and 7beta-hydroxy-DHEA production, effects of CYP inhibitors and of several steroid hormones on DHEA 7-hydroxylation were examined. Using mouse brain microsomes as a source of enzyme, we report now that strong and smaller inhibitions of DHEA 7alpha-hydroxylation were obtained with ketoconazole and alpha-naphthoflavone, respectively, and that neither changed DHEA 7beta-hydroxylation. Metyrapone and antipyrine also inhibited 7alpha-hydroxylation, but by contrast, significantly increased 7beta-hydroxylation of DHEA. This indicated that at least, two different CYPs were responsible for 7alpha- and 7beta-hydroxylation of DHEA. Steroids sharing a 3beta-hydroxylated structure with DHEA, namely pregnenolone, 5-androstene-3beta,17beta-diol and 3beta-hydroxy-5alpha-androstan-17-one, were strong inhibitors of DHEA 7alpha-hydroxylation (non-competitive inhibition with pregnenolone, Ki=2.0 +/- 0.3 microM). In contrast, 7beta-hydroxylation yields were not decreased by the 3beta-hydroxysteroids tested. Moderate inhibition of 7alpha- and 7beta-hydroxylation was obtained with 3-oxosteroids, namely testosterone, progesterone, corticosterone and 4-androsten-3,17-dione. Taken together, these data indicate specific inhibition patterns of DHEA 7alpha- and 7beta-hydroxylation by CYP inhibitors and steroid hormones in mouse brain microsomes and may be used as criteria necessary for identification of the responsible CYP species.

Animals↗

PubMedDate not supplied

Hydroxylation of pregnenolone at the 7 alpha- and 7 beta- positions by mouse liver microsomes. Effects of cytochrome p450 inhibitors and structure-specific inhibition by steroid hormones.

Hydroxylations of pregnenolone (PREG) at the 7 alpha-and 7 beta-positions have been reported in numerous murine tissues and organs, including liver, and the responsible cytochrome P450 (P450) species await identification. Using thin-layer chromatography and gas chromatography-mass spectrometry and crystallization to constant specific activity, we report identification of 7 alpha-hydroxy-PREG and 7 beta-hydroxy-PREG metabolites produced in mouse liver microsomes and kinetic studies of their production with apparent KM values of 2.45 +/- 0.124 microM and 3.41 +/- 0.236 microM for 7 alpha- and 7 beta-hydroxylation, respectively. Investigation of P450 inhibitors and of steroid hormone effects on both 7 alpha- and 7 beta-hydroxylation of PREG showed that 1) different P450 were involved because metyrapone and antipyrine inhibited solely 7 alpha-and 7 beta-hydroxylation, respectively; 2) P450 1A2, 2D6, 2B1, and 2B11 were not responsible for 7 alpha and 7 beta-hydroxylation of PREG because respective specific inhibitors furafylline, quinidine, and chloramphenicol triggered no inhibition; 3) P450 1A1 was responsible for only part of the 7 beta-hydroxylation of PREG because alpha-naphthoflavone, which inhibits specifically P450 1A1, did not suppress entirely 7 beta-hydroxylation while ketoconazole, antipyrine, and metyrapone extensively decreased the 7 beta-hydroxylation; 4) comparison of these findings with those obtained with brain microsomes suggests that tissue-specific P450 species are responsible for the 7 alpha-and 7 beta-hydroxylation of PREG; and 5) 7 alpha-hydroxylation of PREG may be shared with other 3 beta-hydroxysteroids such as isoandrosterone, 5-androstene-3 beta, 17 beta-diol, and dehydroepiandrosterone, which acted in a competitive manner. Taken together, these findings will be of use for identification of the P450 species responsible for 7 alpha- and 7 beta-hydroxylation of PREG and for studies of their activities in liver and other organs.

Animals↗