摘要:
The 3 alpha,5 alpha- and 3 alpha,5 beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone enhance GABAergic neurotransmission and produce inhibitory neurobehavioral and anti-inflammatory effects. Despite Substantial information oil the progesterone derivative (3 alpha,5 alpha)-3-hydroxypregnan-20-one (3 alpha,5 alpha-THP, allopregnanolone), the physiological significance of the other endogenous GABAergic neuroactive steroids has remained elusive. Here, we describe the validation of a method using gas chromatography-mass spectrometry to simultaneously identify serum levels of the eight 3 alpha,5 alpha- and 3 alpha,5 beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone. The method shows specificity. sensitivity and enhanced throughput compared to other methods already available for neuroactive steroid quantification. Administration of pregnenolone to rats and progesterone to women produced selective effects on the 3 alpha,5 alpha- and 3 alpha,5 beta-reduced neuroactive steroids, indicating differential regulation of their biosynthetic pathways. Pregnenolone administration increased serum levels of 3 alpha,5 alpha-THP (+1488%, p<0.001), (3 alpha,5 alpha)-3,21-dihydroxypregnan-20-one (3 alpha,5 alpha-THDOC, +205%, p<0.01), (3 alpha,5 alpha)-3-hydroxyandrostan-17-one (3 alpha,5 alpha-A, +216%, p<0.001), (3 alpha,5 alpha,17 beta)-androstane-3,17-diol (3 alpha,5 alpha-A-diol, +190%, p<0.01). (3 alpha,5 beta)-3-hydroxypregnan-20-one (3 alpha,5 beta-THP) and (3 alpha,5 beta)-3-hydroxyandrostan-17-one (3 alpha,5 beta-A) were not altered, while (3 alpha,5 beta)-3,21-dihydroxypregnan-20-one (3 alpha,5 beta-THDOC) and (3 alpha,5 beta,17 beta)-androstane-3,17-diol (3 alpha,5 beta-A-diol) were increased from undetectable levels to 271 +/- 100 and 2.4 +/- 0.9 pg +/- SEM, respectively (5/8 rats). Progesterone administration increased serum levels of 3 alpha,5 beta-THP (+1806%, p<0.0001), 3 alpha,5 beta-THP (+575%, p<0.001), 3 alpha,5 beta-THDOC (+309%, p<0.001). 3 alpha,5 beta-THDOC levels were increased by 307%, although this increase was not significant because this steroid was detected only in 3/16 control Subjects. Levels of 3 alpha,5 alpha-A, 3 alpha,5 beta-A and pregnenolone were not altered. This method can be used to investigate the physiological and pathological role of neuroactive steroids and to develop bioinarkers and new therapeutics for neurological and psychiatric disorders. (C) 2009 Elsevier Inc. All rights reserved.