Effect of Androstenedione on Adipogenesis in Murine C3H10T1/2 Mesenchymal Cells

Clinical trials of weak androgen androstenedione (AD) administered at a high concentration, showed an increase in muscle mass in men like strong androgens testosterone (T) and dihydrotestosterone (DHT), but did not show any inhibitory effect on fat mass unlike strong androgens. This observation prompted us to check the in-vitro effect of AD on adipogenesis using mouse mesenchymal multipotent cells (C3H10T1/2), which can differentiate into both myoblasts and adipocytes. Results indicated that AD inhibited adipogenesis at 10 nM, 100 nM and 1 μM concentrations, but not at 10 μM concentration. AD did not inhibit adipogenesis at 10 μM concentration and also did not inhibitmyogenesis at 10 μM concentration. Addition of bicalutamide, an androgen receptor (AR) antagonist decreased myogenesis and increased adipogenesis, indicating that the effect of AD was mediated through AR. Another weak androgen dehydroepiandrosterone (DHEA) also showed the same pattern of adipogenesis in 10T1/2 cells. AD also showed a similar pattern of adipogenesis in 3T3-L1 preadipocyte cells. Thus, the in-vitro results of AD on adipogenesis correlated with the in-vivo results of AD on fat-mass from clinical trials and suggested a possible difference in biological action between weak androgens (AD, DHEA) and strong androgens (T, DHT) on adipogenesis. Since the biological action of AD was mediated through AR, this physiological difference onadipogenesis could be due to the nature (partial agonist/antagonist) of AD binding to AR.

Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08

Biotransformation of phytosterol(PS) by a newly isolated mutant Mycobacterium neoaurum ZJUVN-08 to produce androstenedione has been investigated in this paper.The parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology.Androstenedione was the sole product in the fermentation broth catalyzed by the mutant M.neoaurum ZJUVN-08 strain.Results showed that molar ratio of hydroxypropyl-β-cyclodextrin(HP-β-CD) to PS and substrate concentrations were the two most significant factors affecting androstenedione production.By analyzing the statistical model of three-dimensional surface plot,the optimal process conditions were observed at 0.1 g/L inducer,pH 7.0,molar ratio of HP-β-CD to PS 1.92:1,8.98 g/L PS,and at 120 h of incubation time.Under these conditions,the maximum androstenedione yield was 5.96 g/L and nearly the same with the non-optimized(5.99 g/L),while the maximum PS conversion rate was 94.69% which increased by 10.66% compared with the non-optimized(84.03%).The predicted optimum conditions from the mathematical model were in agreement with the verification experimental results.It is considered that response surface methodology was a powerful and efficient method to optimize the parameters of PS biotransformation process.

The Production of Monoclonal Antibody to Androstenedione and the Effect of Passive Immunization on Xinjiang Fine-Wool Sheep With It

Five hybridomas stably secreting monoclonal antibodies (McAbs) to androstenedione were prepared by using artificially synthesized androstenedione-11α-succinyl conjugate with bovine serum albumin (BSA) as antigen. These McAbs showed slightly high cross-reactivity with testosterone (8.1—12.3%) and estrone (0.8—2.5%) and high affinities ranging from 2.0×10~7 to 2.8×10~8 L/M. They were all of the IgG_1 subclass. Xinjiang finewool ewes which were passively immunized with McAbs had higher circulating levels of progesterone (P) and luteinizing hormone (LH) than those of control ewes in two oestrous cycles and these changes led to increased ovulation rate and twin lambs born in young ewes.

Effects of Boesenbergia rotunda （L.） Mansf. on sexual behaviour of male rats

Aim： To study the effects of Boesenbergia rotunda （Krachai） on sexual behaviour in male albino rats. Methods： Thirty-two male Wistar rats were equally divided into four groups： experimental groups were gavaged with the ethanolic extract of the rhizome of B. rotunda at doses of 60, 120 and 240 mg/kg and a control group received distilled water, for 60 days. Sexual behaviour, reproductive organs, diameter of seminiferous tubule, epididymal sperm density, and androgenic hormones were evaluated. Results： Within 30-min observation, there was no significant difference of courtship behaviour, mount frequency （MF）, intromission frequency （IF）, mount latency （ML）, intromission latency （IL）, copulatory efficiency or intercopulatory interval in male rats. In three 10-min intervals over a 30-min period, courtship behaviour and MF during the first 10-min were significantly higher than those in the second and third i0-min observation in all groups, whereas IF had no significant difference. All doses of B. rotunda extract significantly increased the relative testicular weight and the diameter of the seminiferous tubules. The dose of 60 mg/kg also significantly increased the relative weight of the seminal vesicle. Nevertheless, the sperm density, serum testosterone and androstenedione levels were not affected by the B. rotunda extract. Conclusion： B. rotunda does not affect sexual behaviour nor serum androgenic levels.

Synthesis of Novel Chiral 7-Amide Substituted-4-androstene-3,17-dione Derivatives

A series of novel 7-amide substituted-4-androstene-3,17-dione derivatives（8αa ＂8αh or 8βa ＂8βh） was synthesized from the important intermediates 5 by N-acylation and acidic hydrolysis.Compounds 5α and 5β were obtained through the reaction sequence including acetalization,allylic oxidation,oximation and reduction.The structures of the target compounds were characterized by MS,1H NMR,13C NMR and HRMS spectra and their stereo configurations were identified through DEPT（distortionless enhancement by polarization transfer）,HMQC（hetero-nuclear multiple quantum coherence） and NOE（nuclear overhauser effect） correlation.