Administration Of Testosterone Propionate Research Articles

Computer Prediction and Experimental Verification of Guizhi Fuling Pill in the Treatment of Benign Prostatic Hyperplasia

Objective: Benign prostatic hyperplasia (BPH) is a prevalent condition among middle-aged and elderly males. The hypoxia-inducible factor-1α (HIF-1α) / vascular endothelial growth factor A (VEGF-A) signaling pathway plays a key role in proliferative disorders. Guizhi Fuling Pill (GFP) provides significant clinical efficacy in addressing BPH and related hyperplastic ailments. This study aims to elucidate the potential involvement of the HIF-1α / VEGF-A signaling pathway in the therapeutic action of GFP against BPH. Methods: Employing network pharmacology and molecular docking techniques, the mechanism underlying GFP against BPH was anticipated. A rat model of BPH was established to validate the primary targets identified via molecular docking. Subcutaneous administration of testosterone propionate was conducted to induce the BPH rat model for experimental validation. Immunohistochemistry was performed to assess the expression of HIF-1α and VEGF-A in prostate tissue across different experimental groups. Results: The network pharmacology analysis unveiled 111 potential constituents of GFP, with key components such as ent-Epicatechin, quercetin, and Mairin exerting influence on BPH. Essential targets encompass ESR1 and HSP90AA1, with signaling pathways like HIF-1, PI3K-AKT, and VEGF prominently associated with cellular apoptosis, angiogenesis, and proliferation. Molecular docking projections underscored the propensity of core components to interact with key targets, exhibiting favorable binding characteristics. In the context of animal experiments, GFP significantly reduced the expression of HIF-1α and VEGF-A in BPH-afflicted rats as compared to the model group. Conclusion: Collectively, this investigation suggests that GFP’s therapeutic potential for BPH could be attributed to its modulation of multifaceted targets, including HIF-1α and VEGF-A.

Neonatal Aromatase Inhibition Blocked Defeminization of AVPV Kiss1 Neurons and LH Surge-Generating System in Male Rats.

The neuroendocrine system that controls the preovulatory surge of gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH), which triggers ovulation in female mammals, is sexually differentiated in rodents. A transient increase in circulating testosterone levels in male rats within a few hours of birth is primarily responsible for the defeminization of anteroventral periventricular nucleus (AVPV) kisspeptin neurons, which are critical regulators of the GnRH/LH surge. The present study aimed to determine whether neonatal estradiol-17β (E2) converted from testosterone by aromatase primarily causes the defeminization of AVPV kisspeptin neurons and the surge of GnRH/LH in male rodents. The results of the present study showed that the neonatal administration of letrozole (LET), a nonsteroidal aromatase inhibitor, within 2 hours of birth rescued AVPV Kiss1 expression and the LH surge in adult male rats, while the neonatal administration of testosterone propionate (TP) irreversibly attenuated AVPV Kiss1 expression and the LH surge in adult female rats. Furthermore, the neonatal LET-treated Kiss1-Cre-activated tdTomato reporter males exhibited a comparable number of AVPV Kiss1-Cre-activated tdTomato-expressing cells to that of vehicle-treated female rats, while neonatal TP-treated females showed fewer AVPV Kiss1-Cre-activated tdTomato-expressing cells than vehicle-treated females. Moreover, neonatal TP administration significantly decreased the number of arcuate Kiss1-expressing and Kiss1-Cre-activated tdTomato-positive cells and suppressed LH pulses in adult gonadectomized female rats; however, neonatal LET administration failed to affect them. These results suggest that E2 converted from neonatal testosterone is primarily responsible for the defeminization of AVPV kisspeptin neurons and the subsequent GnRH/LH surge generation in male rats.

Scientific Evaluation of A Unani Pharmacopoeia-Based Formulation on BPH in Animal Model

BPH (Benign prostatic hyperplasia) is a common problem of the prostate gland that affects elderly men with age and major cause of lower urinary tract symptoms (LUTS) like urgency of urination, frequency of micturition, dysuria, nocturia, dribbling, and burning of urine. If left untreated, the condition can lead to severe urinary retention, recurrent UTI, and, in severe cases, renal failure, etc. Alpha-blockers and 5-alpha reductase inhibitors (5-ARIs) are two main classes of drugs that are used in conventional therapy to treat BPH. These phytochemical therapies are extremely effective, but standard control methods have unfavorable side effects too. Conversely, evidence, though limited, has shown that alternative medicines are safer and have the potential to improve LUTS and quality of life, in addition to improving sexual dysfunction in patients with BPH. The purpose of the study is to assess the efficacy of 50% hydroalcoholic UPBF extract on BPH in TP-induced rats, and the effects of the drug were compared between the treated and untreated groups. The experimental design for anti-BPH activity was carried out following the Bhavin model. 48 Wistar rats were divided into 6 groups, with 8 rats in each group. Group 1, plane control (untreated rats), group 2, negative control (treated with TP 5 mg/kg), group 3, negative control (treated with TP+ finasteride 1 mg/kg), and groups 4,5 and 6 served as tests controls (A, B, and C) treated with TP+ UPBF extract 100 mg/kg, 150 mg/kg, and 200 mg/kg as low, medium, and high doses, respectively. BPH was induced in rats by administration of testosterone propionate (TP) in arachis oil, and treatment was done orally with a standard drug (finasteride) and a test drug (UPBF) once a day for 28 consecutive days. Before and after initiating the study, each rat was weighed and documented, and after 28 days, the collection of blood samples and the prostate gland was done under light anesthesia. The parameters of the study were accessed, like prostate weight, prostate-to-body weight ratio, histopathological investigation of the prostate, percentage of inhibition, percentage of recovery, and determination of ST and STP levels. Data from the results were expressed as the mean ± standard error of the mean (SEM). Evaluation between group results was carried out with the one-way ANOVA test followed by Tukey’s test, and the significant value of p was <0.05. Both histopathological and biochemical parameter results showed significant improvements in rats treated with finasteride and UPBF extract at a dose of 200 mg/kg (p<0.001). In addition, ST and STP levels showed a significant reduction in comparison with the BPH group. It can be concluded from the findings of the results in both biochemicals and histopathological parameters that the test drug (UPBF) possesses significant effects against the TP-induced BPH in rats. The findings suggested that the test drug can be used independently in the management of BPH as well as for lower urinary tract symptoms.

Orchestrated feedback regulation between melatonin and sex hormones involving GPER1-PKA-CREB signaling in the placenta.

Maintaining placental endocrine homeostasis is crucial for a successful pregnancy. Pre-eclampsia (PE), a gestational complication, is a leading cause of maternal and perinatal morbidity and mortality. Aberrant elevation of testosterone (T0 ) synthesis, reduced estradiol (E2 ), and melatonin productions have been identified in preeclamptic placentas. However, the precise contribution of disrupted homeostasis among these hormones to the occurrence of PE remains unknown. In this study, we established a strong correlation between suppressed melatonin production and decreased E2 as well as elevated T0 synthesis in PE placentas. Administration of the T0 analog testosterone propionate (TP; 2 mg/kg/day) to pregnant mice from E7.5 onwards resulted in PE-like symptoms, along with elevated T0 production and reduced E2 and melatonin production. Notably, supplementation with melatonin (10 mg/kg/day) in TP-treated mice had detrimental effects on fetal and placental development and compromised hormone synthesis. Importantly, E2 , but not T0 , actively enhanced melatonin synthetase AANAT expression and melatonin production in primary human trophoblast (PHT) cells through GPER1-PKA-CREB signaling pathway. On the other hand, melatonin suppressed the level of estrogen synthetase aromatase while promoting the expressions of androgen synthetic enzymes including 17β-HSD3 and 3β-HSD1 in PHT cells. These findings reveal an orchestrated feedback mechanism that maintains homeostasis of placental sex hormones and melatonin. It is implied that abnormal elevation of T0 synthesis likely serves as the primary cause of placental endocrine disturbances associated with PE. The suppression of melatonin may represent an adaptive strategy to correct the imbalance in sex hormone levels within preeclamptic placentas. The findings of this study offer novel evidence that identifies potential targets for the development of innovative therapeutic strategies for PE.

Hepatoprotective potentials of aqueous leaf and seed extracts of Artocarpus heterophyllus on testosterone propionate induced prostatitis in Wistar rats.

This study evaluated the hepatoprotective potentials of aqueous leaf and seed extracts of Artocarpus heterophyllus on testosterone propionate induced prostatitis in Wistar Rat. Male Wistar rats of 100- 125g and 100 in number were obtained from the Department of Pharmacology, University of Port Harcourt animal house and was grouped into 10 of 10 rats each. The various groups were fed and administered with various concentrations (100mg/kg, 200mg/kg, 300mg/kg and 200mg/kg combined extract) of the leaves and seeds extracts of A. heterophyllus. The hepatoprotective potentials was studied by evaluating the liver function parameters on the male Wistar rats by day 21, 42 and 63 of the experiment using standard laboratory methods. The results obtained showed significantly (p< 0.05) increased ALP levels (401.57±0.85) on administration of testosterone propionate in group 2 (induced not treated group) day 21, as against the normal control group (294.57±0.06), while group 5 (Rats induced with TP (4 mg/kg) and administered 200 mg/kg of aqueous extract of A. heterophyllus leaves) revealed minimal level of ALP 293.47±0.006 by day 63. Minimum values for AST 23.00±0.51 was seen in group 8 day 21) with maximum activity 76.82±0.09 seen in group 1 day 21, ALT minimum level 7.67±0.33 was observed in group 4 day 21, while maximum levels 38.41±0.51 was seen in group 2 day 21, Albumin minimum level 17.87±0.09 was seen in group 2 day 63, with maximum level 31.13±0.18 observed in group 8 day 63, while minimum Total Protein 26.31±0.84 was shown in group 2 day 63 and maximum 64.63±0.49 in group 8 day 42. Obtained values from groups 5, 6, 9 and 10 were shown to be significantly different when compared with the negative control in group 2 but not significantly different with control group 1 and 3, hence depicting a more stabilized and hepatoprotective system in the plant extract treated groups compared to the negative control (group 2) at day 21. However, group 5 was shown to have higher hepatoprotective potentials by day 42 followed by group 10.

Testosterone ameliorated the behavioural deficits of gonadectomised rats and counteracted free radicals in a dosage-dependent manner

Testosterone deficiency may induce behavioural changes in individuals. Oxidative stress resulting from a redox imbalance may be implicated in the initiation and progression of neurobehavioural disorders. However, whether exogenous testosterone intervention in male gonadectomised (GDX) rats ameliorates oxidative stress and plays a neuroprotective role remains unknown. Therefore, we examined this hypothesis by performing sham or gonadectomy surgeries on Sprague–Dawley rats with or without supplementation with different doses of testosterone propionate (TP). Open field and Morris water maze tests were performed, the serum and brain testosterone levels, and oxidative stress markers were analysed. GDX and lower TP doses (0.5 mg/kg) induced reduced exploratory and motor behaviours, but impaired spatial learning and memory compared to Sham rats. Administration of physiological TP levels (0.75–1.25 mg/kg) to the GDX rats restored the behaviour observed in the intact rats. However, higher TP doses (1.5–3.0 mg/kg) induced increased exploratory and motor behaviours but impaired spatial learning and memory. These behavioural impairments were accompanied by a marked decrease in levels of antioxidant enzymes (superoxide dismutase and catalase) and an increase in lipid peroxidation levels in the substantia nigra and hippocampus. These findings indicate that TP administration can alter behavioural performance and induce memory and learning impairment, which may result from changes in redox homeostasis in male GDX animals.

Testosterone propionate and Swarna Bhasma treatment modulated D-galactose induced reproductive alterations in male Wistar rats: An experimental study.

The male reproductive system undergoes several adverse age-related changes like decreased hormone synthesis, sperm count, and testicular alteration that can impact on fertility. The study aims to investigate the effects of testosterone propionate (TP), and ayurvedic formulation Swarna Bhasma (SB) on D-galactose (D-gal) induced reproductive aging in male Wistar rats. 60 male Wistar rats were divided into 10 groups of 6 animals. Reproductive aging was induced by D-gal (150 mg/kg Bwt) exposure for 60 days. The rats were then treated by post and combination treatment with TP (2 mg/kg Bwt) and SB (6.75 mg/kg Bwt). Then sperm parameters, reproductive hormones, inflammatory markers, testicular antioxidant enzymes, steroidogenic enzymes, and histological manifestation of testis were evaluated. Exposure of D-gal caused significant (p 0.001) decrease in serum testosterone (T), testicular steroidogenic, and antioxidant enzymes. Administration of TP increased the serum T level, testicular antioxidant enzymes, and spermatogenic profile at a significant level of (p 0.001) compared to D-gal. Further, the SB treatment significantly (p 0.001) elevated the serum T level, sperm count, testicular antioxidant enzymes, steroidogenic enzymes, when compared to D-gal. Both the treatment of TP and SB treatments recovered the reproductive impairments caused by D-gal. However, exogenous T supplementation via TP administration is associated with various side effects during long-term use. SB is an Ayurvedic formulation having a long history of usage in India. The current findings suggest that the SB may be used as a good alternative for potentiating reproductive function in aging males.

In-vivo and in-silico studies revealed the molecular mechanisms of Colocasia esculenta phenolics as novel chemotherapy against benign prostatic hyperplasia via inhibition of 5α-reductase and α1-adrenoceptor.

Benign Prostatic Hyperplasia (BPH) is a major cause of lower urinary tract infections and erectile dysfunction thus a major contributor to lowering the quality of life among older men. In this study, we investigated the molecular mechanism of Colocasia esculenta(CE) as a novel agent for BPH chemotherapy. In vivo, we assigned 45 male Wistar albino rats about 6 weeks old into 9 experimental groups (n = 5). BPH was induced in groups 2-9 with 3 mg/kg of Testosterone Propionate (TP) subcutaneously. Group 2 (BPH) was not treated. Group 3 was treated with 5 mg/kg Finasteride (standard drug). Group 4-9 were treated each with 200 mg/kg body weight (b.w) of CE crude tuber extracts/fractions (ethanol, hexane, dichloromethane, ethyl acetate, butanol, aqueous). At the end of treatment, we sampled the rats' serum to check the level of PSA. In silico, we conducted a molecular docking of the crude extract of CE phenolics (CyP) previously reported, targeting 5α-Reductase and α1-Adrenoceptor linked to the BPH progressions. We adopted the standard inhibitors/antagonists (5α-reductase: finasteride; α1-adrenoceptor: tamsulosin) of the target proteins as controls. Furthermore, the pharmacological properties of the lead molecules were studied in terms of ADMET using swissadme and pKCSM resources, respectively. Results showed that administration of TP in male Wistar albino rats significantly (p < 0.05) elevated serum PSA levels whereas CE crude extracts/fractions significantly (p < 0.05) lowered the serum PSA level. Also, fourteen of the CyPs bind to at least one or two of the target proteins with their binding affinity of between - 9.3 to - 5.6 kcal/mol and - 6.9 to - 4.2 kcal/mol, respectively. The CyPs possess better pharmacological properties compared to the standard drugs. Therefore, they have the potentials to be enlisted for clinical trials towards the management of BPH.

Androgen-Dependent Expression of CUX2 mRNA in the Pig Liver Is Associated with That of Drug Metabolizing Enzymes and Drug Transporters.

We previously identified androgen-dependent sex differences in the mRNA expression of drug metabolizing enzymes (DMEs), including CYPs, sulfotransferases and uridine 5'-diphospho-glucuronosyltransferases, and drug transporters in the pig liver and kidney. To elucidate the mechanism for such sex differences in pigs, we herein focused on the key regulators cut-like homeobox 2 (Cux2), B-cell lymphoma 6 (Bcl6), and signal transducer and activator of transcription 5b (Stat5b), which are reported to be responsible for the sex-biased gene expression of Cyps in the mouse liver. We used real-time RT-PCR to examine androgen-dependent sex differences in the mRNA levels of these regulators in the liver and kidney basically using Meishan and Landrace pigs. Significant sex differences (male > female) in the level of CUX2 mRNA were detected in the liver of both breeds, and levels were significantly decreased in males by castration and increased in castrated males and intact females by administering testosterone propionate. No such clear androgen-dependent sex differences in hepatic BCL6 or STAT5B mRNA expression were observed in either breed. In the kidney, androgen-dependent gene expression of these regulators was not observed. In the liver, CUX2 mRNA expression closely correlated with that of DMEs and drug transporters, which were previously shown to have androgen-dependent expression. Together, these findings demonstrate that hepatic CUX2 mRNA is expressed in an androgen-dependent manner, and strongly suggest that CUX2 plays a key role in the androgen-dependent gene expression of hepatic DMEs and drug transporters.

Sex differences in the hepatotropic effects of antiulcer drugs and placenta cryoextract in an experimental rat liver injury model

Background/Aim: Sex-related variances in drug metabolism provide a foundation for refining treatment protocols for prevalent conditions based on the patient's sex. Tailoring treatment strategies based on sex is particularly noteworthy among patients with comorbid illnesses due to the potential for drug interactions and the impact of concurrent diseases on clinical outcomes. Aim of this study was to assess the hepatotropic effects of antiulcer drugs (esomeprazole, clarithromycin and metronidazole - E/C/M) and placenta cryoextract (CEP) within a simulated model of tetrachloromethane (CCl4 )-induced hepatitis combined with underlying ethanol-induced liver cirrhosis (EILC), with a focus on the role of subjects' sex. Methods: Using 112 male and female rats, the research explored the effects of different sex hormone levels. Chronic EILC was induced by administering a 50.0 % CCl4 oil solution (8 mL/kg) twice a week, combined with a 5.0 % ethanol solution, over 45 days. Total protein (TP) levels and alkaline phosphatase (AP) activity were measured spectrophotometrically. Results: The research findings indicate that the onset of EILC and the administration of E/C/M resulted in a significantly greater 10.8 % (p = 0.03) reduction in TP levels among females compared to males, without altering hormonal status. Introducing CEP led to a noteworthy (p &lt; 0.001) rise in TP levels, by 30.8 % in males and 33.9 % in females, in the context of EILC and E/C/M administration, while maintaining hormonal status. Among male rats, the most elevated AP activity was observed with excess testosterone propionate administration (5.0 [5.0; 5.9] mmol/L), while the lowest level was recorded in rats after testectomy, measuring 3.8 [2.5; 4.7] mmol/L, exhibiting a significant 20.8 % decrease (p &lt; 0.05) compared to male rats without hormonal status changes. In female rats, the study revealed that against the backdrop of EILC and E/C/M administration, the highest AP level was seen in ovariectomised females, reaching 5.8 [5.1; 6.2] mmol/L, reflecting a substantial 9.4 % increase compared to rats without hormonal status changes. Conclusions: The administration of CEP under similar experimental conditions led to the recovery of the liver's protein-synthesising function in both male and female rats. When female sex hormones were introduced to sham-operated female rats, a significant 20.8 % greater reduction in AP levels was observed. Additionally, gonadectomy led to a more pronounced decrease in this enzyme's levels in male rats compared to female rats, indicating the cytoprotective properties of female sex hormones.

Cleistanthins A and B Ameliorate Testosterone-Induced Benign Prostatic Hyperplasia in Castrated Rats by Regulating Apoptosis and Cell Differentiation.

Background The aging male population is at higher risk for benign prostatic hyperplasia (BPH) wherein increased proliferation of stromal and epithelial cells of the prostate is observed. In this study, we investigated the effect of cleistanthins A and B on the inhibition of testosterone-induced BPH in castrated rats. Methodology Male Wistar rats were divided into eight groups (n = 6) and surgical castration was performed. BPH was induced by the administration of testosterone propionate in corn oil at 5 mg/kg for four weeks. The control group received corn oil, and the model group received testosterone propionate. The standard treatment group received finasteride orally along with testosterone. Cleistanthins A and B at 0.3, 1, and 3 mg/kg were administered by oral gavage along with testosterone. After four weeks, rats were sacrificed, and prostates were weighed and assessed for histomorphological, inflammatory, apoptotic, and proliferative markers. Results Cleistanthins A and B decreased prostatic enlargement and histopathological abnormalities. Elevated serum dihydrotestosterone levels were lowered significantly in both the cleistanthin A andcleistanthin B groups compared to the BPH model group. Cleistanthins A and B significantly lowered the serum interleukin (IL)-1β and tumor necrosis factor-alpha inflammatory markers in the test groups. Western blot analysis revealed cleistanthin A downregulated the IL-6, signal transducer and activator of transcription 3/cyclin D1 signaling pathway. Both cleistanthins A and B upregulated the apoptotic markers caspase-3 and cleaved caspase-3, whereas the cell proliferation markers cyclin D1 and proliferating cell nuclear antigen were found to be downregulated. Conclusions Both cleistanthins A and B inhibited BPH in a rat model by apoptotic induction and impeded cell proliferation.

Modeling of Benign Prostatic Hyperplasia in Rats with a High Dose of Testosterone

In order to optimize the testosterone model of benign prostatic hyperplasia, we studied the effect of castration and different doses of testosterone on the induction of the proliferative process in the prostate of Wistar rats. It was shown that 4-week subcutaneous administration of testosterone propionate in a dose of 20 mg/kg causes pronounced proliferative and hemodynamic disorders in the dorsolateral gland morphologically similar in castrated and non-castrated males. Administration of testosterone in a dose of 3 mg/kg had no significant effect on the dynamics of the pathological process in non-operated rats and normalized the structure of the gland in castrated animals. Morphological study showed that castration of males provides no visible advantages in reproducing the testosterone model of benign prostatic hyperplasia. The proposed non-traumatic modification of the model with a high dose of testosterone has good reproducibility and sensitivity to therapeutic agents, as shown by the example of finasteride.

Changes in Gut Microbiome upon Orchiectomy and Testosterone Administration in AOM/DSS-Induced Colon Cancer Mouse Model.

Sex hormones are known to affect the gut microbiota. Previously, we reported that endogenous and exogenous testosterone are associated with colorectal cancer (CRC) development and submucosal invasion. In the present study, we investigated whether the gut microbiota is affected by orchiectomy (ORX) and testosterone propionate (TP) administration using an azoxymethane/dextran sulfate sodium (AOM/DSS)-induced CRC mouse model. Gut microbiota was evaluated by means of 16S rRNA gene sequencing of stool DNA extracted from feces that were obtained at 13 weeks after AOM injection (from 22-week-old animals) and stored in a gas-generating pouch. The increase in microbial diversity (Chao1 and Phylogenetic Diversity index) and Firmicutes/Bacteroidetes (F/B) ratio upon AOM/DSS treatment in ORX mice was significantly decreased by TP supplementation. The ratio of commensal bacteria to opportunistic pathogens was lower in the TP-administered females and ORX mice than in the AOM/DSS group. Opportunistic pathogens (Mucispirillum schaedleri or Akkermansia muciniphila) were identified only in the TP group. In addition, microbial diversity and F/B ratio were higher in male controls than in female and ORX controls. Flintibacter butyricus, Ruminococcus bromii, and Romboutsia timonensis showed similar changes in the male control group as those in the female and ORX controls. In conclusion, testosterone determines the dysbiosis of gut microbiota, which suggests that it plays a role in the sex-related differences in colorectal carcinogenesis.

Testosterone propionate improves motor alterations and dopaminergic damage in the reserpine-induced progressive model of Parkinson's disease

Parkinson's disease (PD) is a chronic and progressive neurodegenerative disorder with a higher susceptibility to occur in men. Studies suggest that this susceptibility is related to the hormonal differences observed between men and women, being a risk factor for PD. In addition, testosterone supplementation has shown controversial results in animal models of PD and parkinsonian patients. This study evaluated the effect of chronic administration of testosterone propionate (TP) on motor behavior and neurochemical parameters in the reserpine-induced rat model of parkinsonism. Male Wistar rats received 15 injections of reserpine (RES – 0.1 mg/kg) every other day and were concomitantly treated with different doses (0.1, 1.0, or 5.0 mg/kg) of daily TP for 30 days. The rats were euthanized 48 h after the 15th injection of RES or vehicle. Brains were removed and subjected to Tyrosine hydroxylase (TH) immunohistochemistry. TP at 1.0 mg/kg reduced the damages caused by reserpine in the vacuous chewing and tong protrusion behaviors and prevented dopaminergic damage in the SNpc, VTA, and Striatum. TP at 5.0 mg/kg reduced the damages caused by reserpine in the catalepsy and tong protrusion behaviors, prevented the weight loss, and prevented dopaminergic damage in the VTA. Our results suggest that chronic administration of TP has a protective effect in a rat model of parkinsonism, improving motor alterations and dopamine depletion induced by RES.

Energy-Resolved Fragmentation Aiding the Structure Elucidation of Steroid Biomarkers.

The identification and confirmation of steroid sulfate metabolites in biological samples are essential to various fields, including anti-doping analysis and clinical sciences. Ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) is the leading method for the detection of intact steroid conjugates in biofluids, but because of the inherent complexity of biological samples and the low concentration of many targets of interest, metabolite identification based solely on mass spectrometry remains a major challenge. The confirmation of new metabolites typically depends on a comparison with synthetically derived reference materials that encompass a range of possible conjugation sites and stereochemistries. Herein, energy-resolved collision-induced dissociation (CID) is used as part of UHPLC-HRMS/MS analysis to distinguish between regio- and stereo-isomeric steroid sulfate compounds. This wholly MS-based approach was employed to guide the synthesis of reference materials to unambiguously confirm the identity of an equine steroid sulfate biomarker of testosterone propionate administration.

Comparative application of testosterone undecanoate and/or testosterone propionate in induction of benign prostatic hyperplasia in Wistar rats.

Testosterone undecanoate is a hormone agent with long-acting potential and is used for testosterone replacement therapy for hypogonadism. This study was designed to investigate application of testosterone undecanoate in maintaining high androgen levels for inducing benign prostatic hyperplasia more conveniently than that for testosterone propionate. We conducted two-part studies to determine the optimal dosage and dosing cycle for efficient and stable induction of benign prostatic hyperplasia using testosterone undecanoate. In the injection dosage substudy, single testosterone undecanoate dose (125, 250, 500, 750, or 1000 mg/kg body weight) was administered, and the optimal concentration was determined for 8weeks by measuring changes in testosterone, dihydrotestosterone, and 5-alpha reductase levels. And then, testosterone undecanoate was administered at the optimal dose at intervals of 1, 2, 3, or 4 weeks for 12weeks to induce benign prostatic hyperplasia. The injection dosage substudy showed dose-dependently higher and more stable levels of testosterone in groups administrated testosterone undecanoate than in groups administered testosterone propionate. In the injection cycle substudy, testosterone undecanoate-administered group stably maintained high levels of testosterone, dihydrotestosterone, and 5-alpha reductase compared with testosterone propionate-administered group for the same injection cycle; moreover, the prostate measurements, an important sign of benign prostatic hyperplasia, were significantly increased. Based on these two substudies, we determined the optimal conditions for inducing benign prostatic hyperplasia stably and more conveniently than that for testosterone propionate. This study suggests an extended application of testosterone undecanoate for inducing benign prostatic hyperplasia that can improve research reliability considering the half-life of testosterone as well as injection dosage and concentration.

Effect of testosterone administration on the eating and mating behaviours of female albino rats

The study investigated the effect of testosterone administration on the eating and mating behaviour of female albino rats. Nine (9) female Wister (albino) rats and three (3) stimulus male albino rats were obtained from the Faculty Pharmacy, University of Uyo, Uyo. The female albino rats weighed 165 to 215 g before administration of testosterone, and 173 to 235 g during the experimental week. The age range of the rats was from 10 to 12 weeks with a mean age of 11 weeks. A within-subject design was used in the study, with a judgmental/purposive non-probability sampling technique adopted. The females were grouped in threes and labelled group 1, group 2, and group 3 while the 3 males were housed in one cage and labelled accordingly. Before the commencement of the experiment, the rats were allowed to acclimatize for a period of two weeks of which food and waters were given ad libitum. After the first nine days, two female albino rats were sampled from the cages – group 1 (rats B and C), group 2 (rats D and E), and group 3 (rats G and H) for blood samples collection to determine their testosterone levels before administration of testosterone propionate. During the experiment, the females’ rats were administered difference level of testosterone concentration. Group 1 was administered 0.5 ml testosterone concentration (low dose), group 2 received 1 ml testosterone concentration (medium dose), and group 3 received 1.5 ml testosterone concentration (high dose). Thereafter, testosterone analysis was carried out on the blood samples collected from the female rats; testosterone concentration in the rats after administration was greater than testosterone concentration before administration. The paired sample t-test and one-way MANOVA was used to analyse the data generated. Results obtained showed that when testosterone levels of the female albino rat increased following testosterone administration, their mean eating behaviour increased from 36.77 to 85.44, while their mean mating behavior also increased from 34.44 to 44.88. Thus, the H1 paired sample t-test showed a t-value of -3.091 with df of 8. The result was significant with p&lt;0.0005, so H1 was accepted. For H2, t-value was also significant at t = -2.74, p&lt;0.005, df of 8 and also accepted. H3 was also significant and accepted at [f (2, 8) = 21.57; p &lt; 0.05] and [F = (2, 8) 7.706; p &lt; 0.05] respectively. In conclusion, the female albino rats eat more after testosterone administration. They also engaged more in mating behaviour after testosterone administration. Hence, the study recommends that married women who have a sexual desire should use testosterone replacement therapy for better results. This should, however, be used moderately.

Previous short-term use of testosterone propionate enhances muscle hypertrophy in Wistar rats submitted to ladder-based resistance training

To investigate the effects of the previous administration of testosterone propionate (TP) on the morphology of the gastrocnemius muscle of Wistar rats submitted to ladder-based resistance training (LRT). Twenty-eight rats were divided equally into groups: initial control (CI), 4-week TP (CT4), 4-week TP + LRT (TRT), and placebo + LRT (RT). The rats from the CT4 and TRT groups were treated with TP for four weeks (10 mg/kg/week). TRT and RT trained for ten weeks. The rodents were euthanized at the end of the experiment, and gastrocnemius muscle, prostate, and left and right testicles were collected. There was no statistical difference between the RT and TRT for final volume load. The prostate mass of the TRT and RT groups was statistically heavier than the CT4 group (P < 0.01). The TRT group's prostate/body mass ratio was statistically different from the CT4 group (P < 0.05). The TRT group was shown to have larger type I, type II, and mean fCSA fibers than all other groups (P < 0.001). Regarding the nuclei/fiber ratio (N/f), the CT4, RT, and TRT groups had higher values than CI (P < 0.01). In addition, the RT group showed a higher N/f ratio than CT4 (P < 0.001) but lower than TRT (P < 0.001). In conclusion, short-term TP administration before resistance training can elicit a greater N/f ratio and size of the mean fCSA of the Gastrocnemius muscle of young adult Wistar rats than resistance training alone.

Blepharis persica increases testosterone biosynthesis by modulating StAR and 3β-HSD expression in rat testicular tissues

Objective: To evaluate the effect of methanolic extract and ethyl acetate fraction of methanol extract prepared from the seeds of Blepharis (B.) persica on testosterone biosynthesis and also to elucidate the underlying mechanism.Methods: Forty-eight male Wistar rats were divided into eight groups (n=6 per group). Group I received 0.3% w/w gum acacia suspension p.o. and served as the normal control group. Group II was administered testosterone propionate in arachis oil i.m. as the positive control group. Group III to V received B. persica methanolic extract p.o. at doses of 50, 100 and 200 mg/kg body weight. Group VI to VIII received B. persica ethyl acetate fraction p.o. at doses of 50, 100 and 200 mg/kg body weight. The testis was used for biochemical estimation and histological studies. The effects of methanolic extract and ethyl acetate fraction of B. persica on testicular testosterone, mRNA expression corresponding to steroidogenic acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) along with 3β-HSD enzyme assay were evaluated in testicular tissues and sperm concentration. Ethyl acetate fraction of B. persica was subjected to column chromatography. In-vitro studies were performed using TM3 cell line at three dose levels (50, 100, 200 μg/mL), each for methanolic extract, ethyl acetate fraction and 2-benzoxazolinone for evaluation of their comparative effect on testosterone production.Results: Ethyl acetate fraction and methanolic extract of B. persica could elevate the testicular testosterone content compared to the normal control group. The treatment with methanolic extract and ethyl acetate fraction of B. persica increased the expression of mRNA corresponding to StAR by 6.7 fold and 10.6 fold, respectively, whereas the mRNA expression of 3β-HSD increased by 5.7 fold and 7.3 fold, respectively. Moreover, fraction and extract treatment exhibited increased 3β-HSD activity in the testicular tissues and were found to elevate sperm concentration in seminal fluid. The spermatogenic potential was further ensured by histological observations. 2-benzoxazolinone was isolated from ethyl acetate fraction and identified using spectral studies. It showed the ability to increase the testosterone content in the TM3 Leydig cells.Conclusions: Methanolic extract and ethyl acetate fraction of B. persica are able to increase the testicular testosterone in rats by elevating mRNA expression of StAR and 3β-HSD in testicular tissues, leading to increase the sperm concentration.

In vivo Administration of Testosterone Propionate in Cattle Analyzed by High Performance Liquid Chromatography-Tandem Mass Spectrometry: An Enzymatic Hydrolysis Study and Drug Abuse Issues

This study involved an animal experiment in which testosterone propionate was intramuscularly injected into four steers, and the ensuing urine samples were analyzed using high-performance liquid chromatography-tandem mass spectrometry. The enzymatic hydrolysis conditions were analyzed using analysis of variance (ANOVA) and response surface methodology employing a 33 Box-Behnken design. A survey study was performed in which 48 urine samples were collected from eight untreated steers to estimate the endogenous levels of testosterone metabolites, and it was found that 17β-testosterone was not detected in the majority of the samples, whereas 17α-testosterone was present in 43 of the 48. Drug abuse was identified within 11 days of 17β-testosterone administration. The ratio between both isomers was also assessed; however, a cut-off limit based on an untreated population could not be estimated due to the absence of the beta isomer. Therefore, there may be a correlation between the exogenous use of testosterone in castrate animals and high levels of 17β-testosterone, although confirmation through further investigation would be required.