All culture media and their ingredients were obtained from Gibco (Life Technologies Ltd, Paisley, UK). Human liver cancer HepG2 cells were cultured in MEM (supplemented with 5% FCS, 1% penicillin/streptomycin, 1% L-glutamine) and maintained in humidified atmosphere at 37°C and 5% CO2. Prior to androgen treatment, the HepG2 cells were split and plated in 12-well plates and pre-incubated for 2–3 days. Testosterone enanthate from Sigma-Aldrich Chemie GmbH and nandrolone decanoate were diluted in ethanol 99.5% from Kemetyl (Haninge, Sweden) (stock solution of 1 mM) and added to the cells for 2–48 h at various concentrations (0.01–10 μM). Other compounds were diluted in ethanol for incubation with cells at a final concentration of 1 μM during 2 h; free testosterone and free nandrolone from NMI, estradiol and estradiol cypionate from Sigma-Aldrich Chemie GmbH and R1881 was kindly provided by Professor Anders Bjartell, Karolinska Institute. The non-treated controls were incubated with vehicle only. The experiments were performed in at least four independent experiments. For RNA experiments the cells were harvested with Trizol from Invitrogen (Paisley, UK) and kept at −80°C.
Testosterone enanthate
Manufactured by Merck Group
Sourced in United States
Testosterone enanthate is a synthetic testosterone ester used for clinical and research purposes. It is a long-acting intramuscular injection that slowly releases testosterone into the body. Testosterone enanthate is used to treat conditions associated with testosterone deficiency.
Automatically generated - may contain errors
Lab products found in correlation
Trizol, by Thermo Fisher Scientific (1 mentions)
5 bromo 2 deoxyuridine brdu, by Merck Group (1 mentions)
Fk506, by Merck Group (1 mentions)
Celltracker green 5 chloromethylfluorescein diacetate, by Thermo Fisher Scientific (1 mentions)
Bicalutamide, by Merck Group (1 mentions)
Cyproterone, by Merck Group (1 mentions)
Cyclosporine a csa, by Merck Group (1 mentions)
Enzyme immunoassay kit, by Cayman Chemical (1 mentions)
Metformin, by Merck Group (1 mentions)
Trans anethole, by Merck Group (1 mentions)
Millex samplicity filter, by Merck Group (1 mentions)
N hexane, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
6 protocols using testosterone enanthate
1
Androgen Treatment Effects on HepG2 Cells
2
Reagents and Treatments for Cell-based Assays
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The following reagents were purchased: Testosterone enanthate, FK506, cyclosporine A (CsA), 5-bromo-2-deoxyuridine (BrdU), bicalutamide, and cyproterone from Sigma-Aldrich (St. Louis, MO, USA); 1-azakenpaullone (1-Azk), 11R-VIVIT, PD98059, LY-294002, and Akt-inhibitor VIII from Calbiochem (La Jolla, CA, USA); CellTracker Green (5-chloromethylfluorescein diacetate) from Molecular Probes/Thermo Fisher Scientific (Eugene, OR, USA); and [3H]-leucine from NEN Radiochemicals Perkin Elmer (Waltham, MA, USA). All other reagents were of analytical grade and commercially available. Testosterone was diluted in absolute ethanol, the final concentration of which was 0.01% (v/v) in the stimulation medium; at this concentration, ethanol exerts no effect on biochemical determinations [10 (link)].
3
Testosterone Enanthate and Antioxidant Effects
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Drugs and materials were obtained as follows:
Testosterone enanthate (IM) (Caspian, Iran), transanethole (Sigma, China), metformin (Sigma, India), and
olive oil (Farabi, Iran). Enzyme immunoassay kits were
used for measurement of insulin (Cayman Chemical,
USA) by ELIZA (14 (link)) and materials for measurement
of oxidative stress marker were obtained from Merck
(Germany). All chemicals and reagents for histological
assessments were purchased from Farzan Azma (Iran).
Testosterone enanthate (IM) (Caspian, Iran), transanethole (Sigma, China), metformin (Sigma, India), and
olive oil (Farabi, Iran). Enzyme immunoassay kits were
used for measurement of insulin (Cayman Chemical,
USA) by ELIZA (14 (link)) and materials for measurement
of oxidative stress marker were obtained from Merck
(Germany). All chemicals and reagents for histological
assessments were purchased from Farzan Azma (Iran).
4
Sensitive LCMS Testosterone Quantification
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Testosterone, testosterone isocaproate, testosterone enanthate, testosterone benzoate, testosterone cypionate, testosterone undecanoate, testosterone valerate, testosterone laurate, testosterone caproate, testosterone hexahydrobenzoate, and testosterone undecenoate were all purchased from Sigma Aldrich (Taufkirchen, Germany); the cortisol-D4 internal standard was supplied by Labco Ltd., UAE. Methanol, hexane, formic acid, deionised water, and LC–MS-grade water and acetonitrile were purchased from Emirates Scientific and Technical Supplies L.L.C. (ESTS) (Dubai, United Arab Emirates). An Ascentis Express F5 column (150 mm × 2.1 mm × 2.7 μm), plastic bags, glass test tubes (5 mL), and glass Pasteur pipettes were received from Gomet (Al Ain, United Arab Emirates). Millex Samplicity TM Filters, 0.20 μm hydrophilic polytetrafluoroethylene (Merck Millipore, Carrigtwohill, Ireland, cat. no. SAMPLG001), and 2 mL glass chromatographic vials with caps (2 mL) were purchased from Emirates Scientific Supplies Ltd., Abu Dhabi, United Arab Emirates.
5
Testosterone-Induced Hair Growth Protocol
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Under anesthesia of ketamine and xylazine (ketamine:xylazine =100:10 mg/kg, intraperitoneal injection), the fur on the animal’s back was removed from the scapular line to the root of the tail, including the thighs of the hind limbs, using an electric clipper, followed by a 2-minute application of ~2 mL of depilation lotion (Nair, Church & Dwight Co., Inc., Ewing, NJ, USA) on the skin. The applied lotion was thoroughly removed with saline swabs. Then, testosterone enanthate (1648004, Sigma, St. Louis, MO, USA, suspended in sesame oil) was intramuscularly injected into the thigh muscle at a dose of 50 mg/kg, twice weekly (with a 3-day gap between injections) for 5 weeks. testosterone enanthate was chosen due to its longer pharmacological half-life (i.e., 4-5 days) than other testosterone-based agents that require more frequent injections [22 ]. The side of the injection was randomized and balanced.
6
Deuterated Surfactant and Steroid Analysis
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Materials Protiated sodium dodecyl sulfate (denoted here as h 25 SDS), testosterone enanthate (TE), D 2 O, n-hexane, ethanol and chloroform were obtained from Sigma-Aldrich (Poole, UK) and were of the highest grade available. The chemical structures of h 25 SDS and TE are shown in Figure 1. Tail-deuterated SDS (described as d 25 SDS) was supplied by the ISIS Oxford Isotope Facility (Didcot, UK). Decon 90 was from Decon Laboratories Limited, (Hove, UK). Ultrapure water, with a resistivity of 18.2 MΩ.cm at 298 ± 0.1 K was from a Milli-Q® Ultrapure Water System (Watford, UK).
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