Characterisation of androstenone metabolism in pig liver microsomes.

Androstenone (5 alpha-androst-16-en-3-one) is a steroid pheromone produced in the testis. Excessive accumulation of androstenone together with skatole (3-methyl-indole) in the adipose tissue of some male pigs leads to "boar taint". In isolated pig hepatocytes androstenone represses the expression of cytochrome P450IIE1 (CYP2E1), the enzyme principally responsible for skatole metabolism. Androstenone can be metabolised in liver microsomes but the pathway has not been established. We have investigated androstenone metabolism in liver microsomes from two breeds of pigs exhibiting low and high levels of androstenone in adipose tissue-Large White (LW) and Meishan (M), respectively. Androstenone was reduced in isolated liver microsomes mainly to beta-androstenol using NADH as a co-factor. The rate of beta-androstenol formation in the presence of NADPH was very low. In microsomes from LW pigs the rate of beta-androstenol formation from androstenone was six times higher than in M pigs. 3beta-hydroxysteroid dehydrogenase (3beta-HSD) was investigated as a likely candidate for the enzyme catalysing androstenone reduction in pig liver. RT-PCR analysis showed that there was no sequence difference in the cDNA encoding 3beta-hydroxysteroid dehydrogenase from LW and M pigs. However, competitive RT-PCR analysis showed that the expression of 3beta-hydroxysteroid dehydrogenase mRNA was about 12 times higher in the case of LW compared to M pigs. It is concluded that the rate of androstenone metabolism in pig liver microsomes is determined by the level of expression of hepatic 3beta-hydroxysteroid dehydrogenase. The differential expression of this enzyme could be a factor affecting the rate of hepatic androstenone metabolism which in turn may influence the level of hepatic CYP2E1 expression and hence the rate of hepatic skatole metabolism.