Testicular Steroidogenic Enzymes Research Articles

P-636 Effects of maternal circadian rhythm changes: intergenerational influences on testicular steroidogenesis in offspring

Abstract Study question Do alterations in the maternal circadian rhythm during pregnancy and lactation result in changes to the male offspring’s testicular steroidogenesis intergenerationally? Summary answer The expression of proteins associated with steroidogenesis is altered due to maternal circadian rhythm changes during pregnancy and lactation periods. What is known already Circadian rhythms impact diverse physiological processes, including reproduction. Several clock genes have been linked to reproductive function. However, there is a complex interaction between hormones, fertility, and the circadian clock. It has been shown that disruption of the circadian clock machinery has negative effects on reproductive biology. Current literature indicates a potential relationship between the circadian clock, testicular steroidogenic-related gene expression profiles, and testosterone synthesis. Steroidogenesis involves a complex cascade of enzymatic reactions. An increasing body of evidence from animal studies suggests a relationship between maternal chronodisruption and long-term offspring health and fertility. Study design, size, duration Male offspring from pregnant Wistar rats (n = 60) were exposed to control (12 hours light/12 hours dark), short day (SD) (8 hours light/16 hours dark), or long day (LD) periods (16 hours light/8 hours dark) during only pregnancy, only lactation, or both periods (n = 6/group) categorized as control, SD pregnancy (SD-P), SD lactation (SD-L), SD pregnancy and lactation (SD-P+L), LD pregnancy (LD-P), LD lactation (LD-L), LD pregnancy and lactation (LD-P+L) groups. Participants/materials, setting, methods The expression of critical components of testicular steroidogenesis, including steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (cytochrome p450scc), 17-beta hydroxysteroid dehydrogenase 3 (HSD17B3), androgen receptor (AR), and luteinizing hormone receptor (LHR), was analyzed in the testis by counting the percentage of positively-stained Leydig cells and ImageJ analysis of immunohistochemistry and western blot. Serum FSH, LH, and testosterone hormones were measured at four zeitgeber time (ZT) points (ZT0, ZT6, ZT12, and ZT18) using ELISA. Main results and the role of chance All of these testicular steroidogenic enzymes were localized in the Leydig cell, and AR was predominantly observed in the Sertoli cell. Although there are no significant differences between the groups in terms of immunolocalization patterns, the expressions of p450scc and AR proteins were significantly decreased in the SD-P and SD-L groups compared to the control group (p < 0.001). In addition, western blot data indicated a decreased expression of p450scc in the SD-P+L group. However, an increased expression of HSD17B3 was detected in both SD-P+L and LD-P+L groups compared to the control group (p = 0.003, p < 0.001, respectively). Among the steroidogenesis-related proteins evaluated, the protein expression levels of StAR and LHR proteins were not altered. FSH showed a statistically significant increase in the LD-L (ZT0), SD-P+L (ZT12), LD-L (ZT12), and LD-P+L (ZT18) groups when compared to the corresponding control groups at the respective time points (p < 0.05). In contrast, LH levels decreased significantly in the LD-P+L (ZT0) group (p < 0.05). Nevertheless, no statistical difference was determined for testosterone levels between groups. Limitations, reasons for caution Most studies on circadian rhythm have used rodent models. Results from animal models are often informative. However, there is a limitation in the appropriate translation of findings in rodents to human studies. Wider implications of the findings The effects triggered by maternal chronodisruption can be intergenerationally transmitted, potentially affecting offspring health. Our results indicate alterations in the expression of proteins involved in testicular steroidogenesis and time-point-dependent changes in the FSH and LH hormone levels due to maternal circadian rhythm changes, further supporting the potential implications for offspring. Trial registration number This work was supported by research grants from the Scientific and Technological Research Council of Turkey (TUBITAK) (SBAG 219S636) and Akdeniz University, The Scientific Research Projects Coordination Unit (TYL-2018-3580).

Protective Effect of L-Carnitine against Deltamethrin Induced Male Reproductive Toxicity in Adult Rats Exposed at their Prepubertal Stage

The present study aims to investigate the protective role of L-Carnitine (LC) against Deltamethrin-induced testicular toxicity in male rats exposed during their early developmental period. DTM was administered at a dose of 6 mg/kg body weight to the prepubertal rat’s Post-Natal Day (PND) 23 rats through oral route and exposure was continued until they reach PND 90. After completion of the exposure period, male rats were assessed for reproductive endpoints. Deltamethrin exposure caused a significant reduction in testosterone production and decreased the sperm parameters such as sperm count, sperm motility, sperm viability and sperm membrane integrity. Deltamethrin intoxication also reduced the reproductive organs’ weight and testicular steroidogenic enzymes (3β-hydroxysteriod dehydrogenase and 17β-hydroxysteriod dehydrogenase) levels. Further DTM exposure induced the oxidative stress as evidenced by the significant reduction in the superoxide dismutase, catalase, and glutathione levels with significant elevation in the malondialdehyde. DTM also induced the significant sperm DNA damage. In addition, DTM exposure significantly declined the testosterone levels over the control group, indicating the impaired steroidogenesis. In addition to the compromised steroidogenesis DTM exposure also deteriorated the testicular architecture according to the histological observations. On the other hand, therapy with LC (100 mg/kg body weight) improved the biomass of reproductive organs, the features of the sperm, testicular steroidogenesis and testicular antioxidant enzymes levels. Concluding that DTM might disturb the testicular antioxidant level that eventually impaired the reproductive health of rats. However, LC supplementation mitigated the reproductive toxicity through its antioxidant property.

Steroidogenic enzyme gene expression and testosterone production are developmentally modulated by bone morphogenetic protein receptor-1B in mouse testis.

Bone morphogenetic proteins (BMPs) and receptors (BMPR-1A, BMPR-1B, BMPR-2) have been shown to be vital for female reproduction, while their roles in males are poorly described. Our study was undertaken to specify the function of BMPR-1B in steroidogenic enzyme gene expression, testosterone production and reproductive development in male mice, given that Bmpr1b mRNA is expressed in mouse testis and Bmpr1b knockout results in compromised fertility. Male mice were passively immunized for 6 days with anti-BMPR-1B in the presence or absence of exogenous gonadotrophins. We then measured the effects of anti-BMPR-1B on testicular hydroxysteroid dehydrogenase isoforms (Hsd3b1, Hsd3b6, and Hsd17b3) and aromatase (Cyp19) mRNA expression, testicular and serum testosterone levels, and testis and seminal vesicle weight. In vitro testosterone production in response to anti-BMPR-1B was determined using testicular culture, and Leydig cell culture in the presence or absence of gonadotrophins. In Leydig cell culture the contribution of seminiferous tubules and Leydig cells were examined by preconditioning the media with these testicular constituents. In adult mice, anti-BMPR-1B increased testosterone and Hsd3b1 but decreased Hsd3b6 and Cyp19 mRNA. In adult testicular culture and seminiferous tubule conditioned Leydig cell culture, anti-BMPR-1B reduced testosterone, while in normal and Leydig cell conditioned Leydig cell culture it increased testosterone levels. In pubertal mice, anti-BMPR-1B reduced gonadotrophin stimulated seminal vesicle growth. In conclusion, BMPR-1B has specific developmental functions in the autocrine and paracrine regulation of testicular steroidogenic enzyme gene expression and testosterone production in adults and in the development of seminal vesicles during puberty.

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.

Exposure to acrylamide inhibits testosterone production in mice testes and Leydig cells by activating ERK1/2 phosphorylation.

Acrylamide (ACR) is formed during the cooking of starchy foods at high temperatures. Accumulating evidence has shown that ACR has toxic effects, but the mechanism of its potential reproductive toxicity remains unclear. In this study, we observed that ACR caused weight loss in mice. There was no significant difference in the weight of testis and epididymis between the low/medium-dose ACR group and the control group. And the number of epididymal sperms, testicular Leydig cells, serum testosterone level, testicular steroidogenic genes and enzymes, including cytochrome P450 family 11 subfamily A member 1 (CYP11A1) and cytochrome P450 family 17 subfamily A member 1 (CYP17A1), were decreased in the medium/high-dose ACR group. Additional cell experiments showed that the apoptosis rate and the level of reactive oxygen species (ROS) were increased, and testosterone levels and CYP17A1 protein expression were reduced in Leydig cells with treated ACR. Furthermore, the phosphorylation levels of extracellular signal-regulated kinases (ERK1/2) increased significantly; however, there was no significant difference in the levels of serine-threonine protein kinase (AKT) phosphorylation in the testis of mice and Leydig cells treated with ACR. These results suggest that ACR exposure leads to the damage of testicular structure and function and a decline in testosterone synthesis in Leydig cells and mouse testis, which may be related to the activated phosphorylation of ERK1/2.

Arjunolic acid reverses fluoxetine-induced alterations in testicular steroidogenic enzymes and membrane bound ionic pump imbalance through suppression of oxido-inflammatory stress and apoptosis.

The impact of the anti-depressant therapy on gonadal function has been recognized and discussed over the years. However, data to supplement our understanding of the impact of arjunolic acid (AA) therapies in protecting against FXT-induced gonadal dysfunction is lacking clear scientific evidence. Hence, this study aimed to investigate the possible effect of AA on fluoxetine-induced altered testicular function in rats. After 14 days acclimatization, Thirty-six (36) adult male rats were randomly divided into 6 groups (n=6). Rats in groups 1 received normal saline (10mL/kg); groups 2 & 3 were given AA (1.0mg/kg body weight) and AA (2.0mg/kg body weight), respectively; whereas, rats in group 4 were given FXT (10mg/kg/p.o/day), and groups 5 & 6 were given a combination of FXT (10mg/kg) + AA (1.0mg/kg body weight); and FXT (10mg/kg) + AA (2.0mg/kg body weight), respectively. The results shows that FXT significantly altered testicular steroidogenic enzymes (3ß-HSD and 17ß-HSD) and proton pump ATPase (Na+/K+ ATPase, Ca2+ ATPase and H+ ATPase) activities, as well as testicular architecture when compared with controls. More so, FXT caused oxido-inflammation and apoptosis, as evidence by increases in MDA, MPO, TNF-α, IL-1ß, Caspase 3 and p53. However, AA at a different dose significantly ameliorated the destructive impacts of FXT on steroidogenic enzymes, proton pump ATPase as well as increased Bcl-2, SOD, CAT, GSH and improved testicular architecture in rats. AA reverses fluoxetine-induced alterations in testicular steroidogenic enzymes and membrane-bound ionic pump through suppression of oxido-inflammatory stress and apoptosis.

Seasonal change of circulating leptin associated with testicular activities of the wild ground squirrels (Citellus dauricus).

The purpose of this study was to explore the variations in the circulating leptin concentrations of the wild ground squirrels in relation to seasonal changes in testicular activities. Hematoxylin-eosin staining showed all types of elongated spermatids and spermatogenic cells existed in the testis in April, while the primary spermatocytes and spermatogonia were most advanced stages of germ cells in June. In addition, the primary spermatocytes, secondary spermatocytes, and spermatogonia were most advanced stages of germ cells in September. The highest circulating leptin concentration was consistent with the maximum body weight results from accumulation of adipose tissue in September. The mRNA expression level of leptin receptor (Ob-R) and STAT3 was lowest in June, raised in September, and remained increased in April. Ob-R and STAT3 were stronger staining in the Leydig cells in July. Moreover, the concentrations of testosterone (T) showed the maximum values in April, the minimum values in June, and significant increases in September. Furthermore, it is worth noting that the levels of T increased with the mRNA levels of Ob-R, STAT3, StAR, and testicular steroidogenic enzymes (3β-HSD, P450c17, and P450scc). Moreover, RNA-seq analyses of testis during the different periods showed that a total of 4209 genes were differentially expressed genes (DEGs); further analysis revealed that DEGs related with the Jak/STAT pathways and reproduction were altered. Taken together, the results suggested that the leptin regulated testicular function through the Jak/STAT pathways and testicular steroidogenic factor expressions.

Immobilization stress exacerbates arsenic-induced reprotoxic effects in adult rats.

The central objective of this study was to investigate the cumulative effects restraint stress and sodium arsenite on reproductive health in male rats. Healthy male Wistar rats were allocated into 4 groups (n=8). Animals in group 1 served as controls and did not subjected to any stress. Rats in groups 2, 3, and 4 were subjected to either restraint stress (5h/day) or maintained on arsenic (25ppm) via drinking water or both for 65days. After completion of the experimental period, all the rats were analyzed for selected reproductive endpoints. Restraint stress or sodium arsenite treatment increased serum corticosterone levels, reduced testicular daily sperm count, epididymal sperm viability, motility, membrane integrity, and decreased testicular steroidogenic enzymes such as 3β- and 17β-hydroxysteroid dehydrogenases associated with reduced serum testosterone levels, deteriorated testicular architecture, and reduced activity levels of testicular superoxide dismutase and catalase accompanied by elevated lipid peroxidation levels. In rats subjected to restraint stress and sodium arsenite, a significant decrease in selected sperm qualitative and quantitative parameters, serum testosterone levels were observed as compared with rats subjected to sodium arsenite alone. A significant increase in the levels of lipid peroxidation with a concomitant decrease in the activities of antioxidant enzymes was observed in the testis of rats subjected to both restraint stress and sodium arsenite treatment as compared with sodium arsenite alone intoxicated rats. Surprisingly, serum corticosterone levels were significantly elevated in rats following both stressors as compared with arsenic alone treated rats. Analysis of atomic absorption spectroscopy revealed that the accumulation of arsenic in the testis of arsenic-treated and arsenic plus immobilization stress groups was significant as compared with controls. Based on the findings, it can be concluded that deterioration of male reproductive health could be accelerated in arsenic intoxicated rats following restraint stress.

Lutein Attenuates Cyclosporin-induced Testicular Impairment in Male Rats through Modulation of Androgenic Hormones and Enzymes

Background: Male reproductive toxicity has been linked to cyclosporine, a commonly used immunosuppressive drug for the prevention of organ rejection in patients undergoing renal transplant. The goal of this study was to elucidate how lutein protects male testicles from cyclosporine-induced damage. Methods: Thirty adult male Wistar rats were randomly allotted to five groups, each with six animals. Rats in groups 1 and 2 were given saline (2 mL/kg/day p.o) and corn oil (2 mL/day p.o) respectively. Rats in groups 3 and 4 were given lutein (40 mg/kg/day p.o) and cyclosporine (40 mg/kg/p.o./day), while rats in group 5 were given a combination of cyclosporine (40 mg/kg/day p.o) and lutein (40 mg/kg/day p.o). At the end of the fourth week, sperm indices, serum hormones, testicular steroidogenic enzymes [3 and 17 beta-hydroxysteroid dehydrogenases (3ß-HSD and 17ß-HSD)] and enzyme markers of spermatogenesis [lactate dehydrogenase (LDH-X), sorbitol dehydrogenase (SDH), gamma glutamyl transferase (?-GT), acid phosphatase (ACP) and alkaline phosphatase (ALP)] were assayed. The testis of each rat was also investigated for histopathological abnormalities and germ cell count. Results: Lutein attenuated cyclosporine-induced sperm impairment. In rats treated with cyclosporine, lutein reduced LDH-X, SDH, ACP, ?-GT; raised LH, FSH, testosterone, 3ß-HSD, 17ß-HSD, ALP levels, and improved spermatogenesis. Conclusion: These results suggest that lutein attenuates cyclosporine-induced testicular impairment through modulation of androgenic hormones and enzymes.

Assessment of testicular steroidogenic enzymes expression in experimental animal model following withdrawal of nandrolone decanoate.

Anabolic steroids are frequently used to increase the growth rate of meat-producing animals. Exposure to an anabolic-androgenic steroid, nandrolone decanoate (ND), is associated with expressional reduction of testicular steroidogenic enzymes. However, the effect of withdrawal of ND exposure on the expression of these testicular molecules has not been thoroughly explored. The current research investigated expression changes of testicular steroidogenic enzymes in rats at several recovery periods (2, 6, and 12 weeks) after the stop of ND treatment with different doses (2 and 10 mg/kg body weight) for 12 weeks. Body and testis weights were recorded, and transcript levels of molecules were determined by quantitative real-time polymerase chain reaction (PCR). The immunohistochemistry was used to examine the changes of immuno-intensities of molecules. At 6 and 12 weeks of the recovery period, the 10 mg/kg ND-treated rats were lighter than other experimental groups. The interstitial compartment vanished by ND treatment filled up as the recovery period became longer. The expression of steroidogenic acute regulatory protein was returned to the control level at 12 weeks of the recovery period. Expression levels of cytochrome P450 side-chain cleavage and 17a-hydroxylase were increased in 2 mg/kg ND-treated group at 6 weeks of the recovery period, and transcript levels of these molecules in 2 and 10 mg/kg ND-treated groups at 12 weeks of the recovery period were significantly lower than the control. Expression levels of 3β-hydroxysteroid dehydrogenase (HSD) type I and 17β-HSD type 3 in 2 mg/kg ND-treated group were comparable with those of control at 12 weeks of the recovery period, but not in 10 mg/kg ND-treated group. Expression of cytochrome P450 aromatase (Cyp19) was reverted to the control level at 2 weeks of the recovery period. Except for Cyp19, there was a visible increase of immuno-staining intensity of other testicular steroidogenic enzymes in the Leydig cells as the recovery period progressed. This research has demonstrated that the cease of ND administration could restore the expression of testicular steroidogenic enzymes close to the normal level. Nevertheless, a relatively long recovery period, compared to the ND-exposure period would be required to retrieve normal expression levels of testicular steroidogenic enzymes.

Beneficial Role of Ginger Powder (Zingiber officinale) against Acephate-induced Reprotoxicity in Adult Male Rats

Aims: The present study was aimed to investigate the protective role of ginger against acephate-induced testicular toxicity in adult rats.
 Methodology: Rats were allocated into four groups where animals in group I served as controls, while animals in group II, III and group IV were treated as experimental rats. Rats in groups II, III and IV were treated with acephate (50mg/kg body weight), ginger (100mg/kg body weight) and combination of both acephate and ginger, respectively over a period of 60 days. After completion of experimental period sperm count, sperm viability, sperm motility, sperm membrane integrity, testicular steroidogenic marker enzymes (3β-HSD and 17β-HSD, serum testosterone and testicular architecture was performed in both control and experimental rats.
 Results: Relative weights of reproductive organs, sperm count, sperm viability, sperm motility and sperm membrane integrity were significantly decreased in acephate treated rats over controls. Acephate administration also reduced the circulatory levels of testosterone associated with a significant reduction in the testicular steroidogenic marker enzymes (3β-HSD and 17β-HSD) in rats. The testicular architecture was disrupted in acephate intoxicated rats. In contrast, ginger administration significantly recovered the acephate-induced suppressed selected reproductive parameters with increased circulatory levels of testosterone and restoration of sperm endpoints in as compared to acephate alone treated rats. No significant changes were observed in any of the selected reproductive endpoints in ginger treated rats as compared to controls.
 Conclusion: The results can be concluded that supplementation of ginger mitigates the negative effects of acephate on male reproductive health via amelioration of testicular setroidogenesis and spermatogenesis and epididymal sperm maturation events in rats.

Protective effect of myricetin on nonylphenol-induced testicular toxicity: biochemical, steroidogenic, hormonal, spermatogenic, and histological-based evidences

Nonylphenol (NP) is an environmental contaminant, which induces testicular toxicity through oxidative stress. Myricetin (MYR) is a naturally occurring flavonol having powerful antioxidant activity. The current research was planned to examine the ameliorative role of MYR against NP-induced testicular damage. A total of 24 adult male Sprague-Dawley rats were randomly divided into 4 equivalent groups: control (0.1% DMSO), NP group (50mg kg-1), NP + MYR group (50mg kg-1; 100mg kg-1), and MYR-treated group (100mg kg-1). NP administration significantly (p< 0.05) decreased the activity of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GSR), and protein content while significantly (p< 0.05) elevating the thiobarbituric acid reactive substances (TBARS) and reactive oxygen species (ROS) levels. Additionally, NP significantly (p< 0.05) reduced the sperm motility, gene expression of testicular steroidogenic enzymes (3β-HSD, 3β-hydroxysteroid dehydrogenase; 17β-HSD, 17β-hydroxysteroid dehydrogenase; StAR, steroidogenic-acute regulatory protein), level of luteinizing hormone (LH), follicle-stimulating hormone (FSH), plasma testosterone, and daily sperm production (DSP). On the other hand, it raised the testicular cholesterol, dead sperms, and head, midpiece, and tail abnormalities along with abnormal histomorphometry. However, MYR remarkably abrogated NP-induced damages. In conclusion, the outcomes of the study suggest that MYR can effectively alleviate the NP-induced oxidative stress and testicular damages.

Di-n-butyl phthalate diminishes testicular steroidogenesis by blocking the hypothalamic-pituitary-testicular axis: relationship with germ cell apoptosis in Japanese quail.

Although di-n-butyl phthalate (DBP) induces germ cell apoptosis, the underlying mechanism is not yet clear in quail. In this study, prepubertal quails were given a single dose of 500mg kg-1 DBP by gavage and were then killed 3, 6 and 24h after treatment. There was a significant reduction in intratesticular testosterone (ITT) concentrations and testicular steroidogenic enzyme mRNA expression and a significant increase in germ cell apoptosis in DBP-treated compared with control quails at all time points. Maximum apoptosis was detected 6h after treatment and the maximum reduction in testosterone concentrations was at 3h. To investigate whether DBP suppressed testicular steroidogenesis by affecting the hypothalamic-pituitary-testicular axis, we analysed pituitary LH subunit β (Lhb) mRNA expression and serum LH concentrations. At all time points, pituitary Lhb expression and serum LH concentrations were significantly decreased following DBP treatment. The present observations suggest the possibility that DBP blocked LH secretion from the hypothalamus and/or pituitary, thereby decreasing LH stimulation of Leydig cells and reducing ITT concentrations. DBP-induced decreases in ITT concentrations may cause changes to the physical structure of Sertoli cells, which, in turn, may induce germ cell apoptosis.

Recovery of Prenatal Baicalein Exposure Perturbed Reproduction by Postnatal Exposure of Testosterone in Male Mice.

Baicalein (BC), a flavonoid, which lacks the qualities of reproductive health and shows adverse effects, is tested in this study. Inseminated mice were injected with 30, 60, and 90 mg BC/Kg body weight on gestation days 11, 13, 15, and 17. The F1 BC-exposed males of each dosage were divided into six groups. First three groups (n = 6 from each BC dosage) were used for assessment of reproductive performance, the others (n = 4 from each BC dosage) were administered with testosterone 4.16 mg/kg body weight on postnatal days 21, 31, and 41. The reproductive health of adult F1 males at the age of 55 and 60 was tested. Prenatal BC exposure showed reduced fertility after cohabitation with control females. The BC exposure significantly reduced the body weight, tissue indices, and sperm parameters (motility, count, viability, and daily sperm count) and altered the sperm membrane in a hypoosmotic swelling test. A downward trend was observed in testicular steroidogenic marker enzymes (3β- and 17β-steroid dehydrogenases) and serum testosterone, whereas increase in serum titers of FSH and LH along with altered the testicular histology. Conversely, testosterone (4.16 mg/kg body weight) partially recovered reduced male reproductive health by BC. BC impaired male reproductive health due to low levels of testosterone is reverted by external testosterone is evidenced in this study.

Nobiletin ameliorates nonylphenol-induced testicular damage by improving biochemical, steroidogenic, hormonal, spermatogenic, apoptotic and histological profile.

Nonylphenol (NP) is an environmental contaminant, which adversely affects the male fertility due to endocrine disruption and generation of oxidative stress. The current research was planned to assess the effects of nobiletin (NOB), a polymethoxyflavone, on NP-induced testicular damages. Twenty-four male rats were divided into 4 groups: control (0.1% DMSO), NP group (50 mg/kg), NP+NOB group (50 mg/kg + 25 mg/kg), and NOB group (25 mg/kg). Our results revealed that NP brought down the activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GSR), while elevated the level of thiobarbituric acid reactive substances (TBARS). Additionally, NP decreased the level of follicle-stimulating hormone (FSH), luteinizing hormone (LH), plasma testosterone, daily sperm production (DSP), epididymal sperm count, viability, motility, gene expression of testicular steroidogenic enzymes (StAR, 3β-HSD and 17β-HSD) and anti-apoptotic protein (Bcl-2), as well as number of spermatogenic cells belonging to various stages. Whereas, sperm (head, mid-piece/neck and tail) abnormalities, expression of apoptotic proteins (Bax and caspase-3), and histopathological damages were increased. However, NOB remarkably reversed all the damages caused by NP. Therefore, it is deduced that NOB could be used as a potential therapeutic to counter the NP-prompted oxidative stress and apoptotic damages in testes.

Testosterone recuperates deteriorated male fertility in cypermethrin intoxicated rats.

The present study investigates the protective effects of testosterone against reproductive toxicity induced by cypermethrin (50mg/kg body weight) in rats. Significant reduction in the testicular and accessory sex organ weights were observed in cypermethrin-treated rats over controls. Cypermethrin intoxication significantly reduced testicular daily sperm count, epididymal sperm count, sperm motility, sperm viability and HOS-tail coiled sperm accompanied by significant reduction in the activity levels of testicular steroidogenic enzymes such as 3β- and 17β- hydroxysteroid dehydrogenases in rats as compared to controls. Further, qPCR studies indicated that the mRNA expression levels of steroidogenic acute regulatory protein (StAR) significantly decreased in cypermethrin-treated rats over controls. Molecular docking analysis indicated that the binding affinity of cypermethrin (-11.2kcal/mol) towards StAR protein was greater as compared to its natural ligand, cholesterol (-8.2kcal/mol) suggesting improper cholesterol channeling across the testis. Significant reduction in the circulatory levels of testosterone was also recorded in cypermethrin-exposed rats. An increase in pre- and post-implantation loss was observed in rats cohabited with cypermethrin-treated rats. On the other hand, testosterone (4.16mg/kg body weight) treatment ameliorated cypermethrin-induced reprotoxic effects in rats. To conclude, cypermethrin-induced deterioration of suppressed reproductive performance in male rats could be linked to its antiandrogenic effects and on the other hand, testosterone-mediated protection of male reproductive health in cypermethrin-treated rats at least in part occurs via restoration of testosterone biosynthesis, spermatogenesis and sperm maturation events.

SUN-159 17-beta Hydroxysteroid Dehydrogenase 3 Deficiency in 1 Month Old Infant

Background: 17-beta hydroxysteroid dehydrogenase 3 deficiency is a rare autosomal recessive disorder caused by mutations in HSD17B3 encoding the enzyme which converts androstenedione to testosterone. It is characterized in 46, XY males by incomplete virilization, including micropenis and hypospadias. Clinical Case: We report a 1 month old infant who presented with ambiguous genitalia. Prenatal non-invasive screening showed a Y chromosome, however, fetal ultrasound revealed female genitalia. The infant was born with micropenis (~1.4 cm in length) and proximal hypospadius, with enlarged labioscrotal folds and palpable gonads bilaterally. The urethral meatus had been relocated surgically to the glans. There was an apparent vaginal orifice with a normally positioned anus. Initial testing revealed a normal serum 17-OHP (90 ng/dl, n<200 ng/dl) and normal electrolytes. Abdominal US showed normal kidneys. Pelvic US demonstrated no Mullerian structures; gonads thought to be testes were identified in the labioscrotal folds. At 3 months of age, the infant underwent a 3 day HCG stimulation testing with a borderline testosterone response to 132 ng/dl, androstenedione 78 ng/dl and DHT 25 ng/dl. T/A ratio was unremarkable at 1.7 (n>0.8). Thus, hormonal testing was unsupportive of a testicular steroidogenic enzyme deficiency or androgen insensitivity syndrome. Karyotype was confirmed as 46, XY with microarray evidence of multiple regions of homozygosity. Genotyping with a 46, XY DSD panel (GeneDx) revealed a homozygous pathogenic variant c.608 C>T (p.A203V) in exon 9 of the HSD17B3 gene, consistent with a diagnosis of autosomal recessive 17-beta hydroxysteroid dehydrogenase 3 deficiency. Parents are of Arabic descent and are consanguineous. An older brother was also born with ambiguous genital and was later found to be homozygous for the same mutation. This mutation has been identified in the homozygous state in several unrelated affected patients. Previously published functional studies demonstrated loss of enzymatic activity with this missense mutation (1). Male gender was assigned at birth, and parents wish to continue male sex of rearing. Conclusion: Molecular genetic analysis utilizing a commercially available candidate gene panel for 46, XY disorders of sex development diagnosed 17 beta-HSD3 deficiency in this case where hormonal testing was not informative. Early and correct diagnosis is key in planning medical treatment to facilitate pubertal development.

Involvement of dibutyl phthalate on male reproductive toxicity in the freshwater fish Pseudetroplus maculatus (Bloch, 1795)

ABSTRACT The testicular toxicity of dibutyl phthalate (DBP) was evaluated on the freshwater fish Pseudetroplus maculatus. The median lethal concentration of DBP for 96 h determined by probit analysis was 2 mg L−1, and its sublethal (0.2 mg L−1) exposure for 1, 3, 4, 7, and 15 d showed no change in the body weight but affected the reproductive parameters. The concentrations of alkali-labile phosphates and total protein in plasma and testis increased significantly (P < 0.05) with increase in the level of estradiol and decrease in testosterone, illustrating the adverse effect of DBP on male reproductive functions. Impairment in testicular steroidogenic enzymes and reduction in serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), and thyroid stimulating hormone (TSH) with increase in cortisol suggested reproductive dysfunction and stress response of the fish. Further, failure in the reproductive performance was evidenced by reduction in sperm motility, viability, and sperm count. Histological examination confirmed the absence of spermatozoa, completely disorganized spermatogonia, and spermatocytes in treatment groups, suggesting testicular toxicity of DBP, which may contribute to infertility and decline in fish population in the natural environment.

Ameliorative effect of curcumin against lead acetate-induced hemato-biochemical alterations, hepatotoxicity, and testicular oxidative damage in rats.

Lead, toxic heavy metal of global concern, induces toxicity in various organs via oxidative stress. Thereby, in this study, the protective role of curcumin against lead acetate-induced toxicity was evaluated. Thirty-two male albino rats were allocated equally into four groups and orally administered with corn oil as a vehicle (Cont.), curcumin (CUR) (400mg/kg bw), lead acetate (LA) (100mg/kg bw), and lead acetate plus curcumin (LA + CUR). All rats had received their treatments daily for 4weeks. The results revealed that LA toxicity induced normocytic normochromic anemia with significant leukocytosis and lymphocytosis. Moreover, LA-intoxicated rats showed a marked elevation in the liver enzyme activities, serum cholesterol, and triglyceride levels. In contrast, sero-immunological parameters, total protein, albumin, globulin, and testosterone levels were significantly reduced compared to the control rats. Additionally, LA-induced hepatic and testicular oxidative damage revealed by marked increased in MDA level with prominent reduction in the antioxidant system. The gene expression of the hepatic pro-inflammatory markers and testicular steroidogenic biomarkers including LHR and aromatase were significantly upregulated; meanwhile, the expressions of testicular StAR, CYP17a, 3B-HDS, SR-B1, and P450SCC were significantly downregulated in the LA-intoxicated group. Curcumin treatment could partially improve the hematological, biochemical, and histopathological alterations induced by LA. Also, it was observed that curcumin significantly restored hepatic pro-inflammatory markers and testicular steroidogenic enzymes. In conclusion, curcumin has antioxidant, anti-inflammatory, and immunomodulatory effects and is able to minimize the LA-induced oxidative damage in rats.

Embryonic cadmium exposure of male rats alters reproductive functions at adulthood, but without overt alterations in developmental and behavioral outcomes and metabolism

The effects of exposure of pregnant rats to cadmium (Cd) on the developmental and behavioral outcomes, reproductive functions, and metabolism of their male progeny were evaluated. Rats (Wistar) were injected intraperitoneally with either 0.5 or 5.0 µg Cd/kg body weight from day 12 to day 19 of pregnancy. The male offspring were evaluated for their developmental outcomes and behavioral changes. All developmental and behavioral parameters assessed were comparable among the different groups. All male pups were allowed to develop until 100 days of age and evaluated for reproductive end points. The results revealed that although the body weights and relative weights of liver, brain, kidney, testis, and epididymis were not altered, reproductive parameters, including daily sperm production, epididymal sperm numbers, and concentrations of motile, viable, and hypo-osmotic tail-swelled sperm declined significantly in rats exposed to 0.5 and 5.0 µg Cd during embryonic development. In addition, plasma testosterone levels and activity levels of testicular steroidogenic enzymes also decreased in these rats. In the fertility study, although each male in the 0.5, 5.0 µg, and control groups produced a copulatory plug and impregnated a female, the mean number of implantations and live fetuses was reduced significantly in females mated with rats exposed to 0.5 and 5.0 µg Cd during the prenatal period. The general metabolism of the animals exposed to Cd during embryonic development was comparable with the controls as evidenced by no significant changes in the activity levels of succinate dehydrogenase, lactate dehydrogenase, glucose-6-phosphate dehydrogenase, glutamate dehydrogenase, aspartate aminotransaminase, and alanine aminotransaminases in the liver, kidney, and testis. The results thus suggest that maternal Cd exposure during embryonic development markedly affected the spermatogenesis, steroidogenesis, and fertility potential, but without alterations in the development, behavior, and metabolism.