Abstract
Steroids metabolism plays an important role in mammals and contributes to quality of pig meat. The main objective of this study was to identify metabolites of androstenone, 17β-estradiol and dihydrotestosterone using primary cultured pig hepatocytes as a model system. The role of 3β-hydroxysteroid dehydrogenase (3βHSD) in regulation of steroid metabolism was also validated using trilostane, a specific 3βHSD inhibitor. Steroid glucuronide conjugated metabolites were detected by liquid chromatography time of flight mass spectrometry (LC-TOF-MS). 3βHSD enzyme was essential for metabolism of androstenone to 5α-androst-16-en-3β-ol, which then formed androstenone glucuronide conjugate. Metabolism of 17β-estradiol was accompanied by formation of glucuronide-conjugated estrone and glucuronide-conjugated estradiol. The ratio of the two metabolites was around 5∶1. 3βHSD enzyme was not involved in 17β-estradiol metabolism. 5α-Dihydrotestosterone-17β-glucuronide was identified as a dihydrotestosterone metabolite, and this metabolism was related to 3βHSD enzyme activity as demonstrated by inhibition study.
Highlights
Steroid hormones are essential for a range of physiological processes in mammals
A time-course of androstenone metabolism showed that androstenone was non-detectable in the cell culture medium after 24 h of incubation (Figure 1B)
The results demonstrated a sole mass spectrometric peak of m/z2 = 449.2424, with the retention time corresponding to the peak of androstenone metabolite in the cultured medium samples analyzed in electrospray ionization (ESI)– mode
Summary
Steroid hormones are essential for a range of physiological processes in mammals. They act as body chemical messengers, and play critical role in organism development and maturation [1]. Steroids play an important role in regulation of pig metabolism and pig meat quality. Boar taint can be prevented by surgical castration which reduces the level of steroids, including androstenone [6]. Due to the EU initiative to ban surgical castration by 2018, boar taint becomes an increasing issue for international pig meat industry and introduces the challenge of reduction of androstenone level by the means other than castrations [7]. To the best of our knowledge, relationship between 3bHSD and 17bestradiol and dihydrotestosterone metabolism has not been investigated
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