Chronic stress (CS) contributes to male infertility, reduced testosterone levels, and impaired semen quality. CS models induced by glucocorticoids, such as dexamethasone (DEX), negatively affect sperm parameters and testicular health, notably by promoting testicular apoptosis. While individual plant extracts have been studied for their ability to mitigate stress-induced reproductive dysfunction, the preventive effect of the Tri Garn Pis (TGP) polyherbal extract in DEX-induced CS (DexCS) has not previously been investigated. This study evaluated the effects of TGP extract on testicular function, sexual behavior, and sperm quality in DexCS male mice. Seventy-two ICR mice were randomly divided into six groups: control, DexCS, TGP (50, 100, and 200) + DexCS, and TGP200. Mice received TGP (50, 100, 200 mg/kgBW) for 14 days before DEX co-treatment for 28 days. Behavioral and reproductive assessments included depression-like behavior tests, sexual behavior, sperm quality, testicular histopathology, steroidogenesis proteins (AR, CYP11A1, StAR), and apoptosis markers (Hsp70, caspase-3, caspase-9). TGP extract—which is rich in phenolics and flavonoids with antioxidant activity—improved depressive behavior, sexual performance, testicular histology, and low sperm quality. TGP also upregulated testicular StAR expression while reducing caspase-3 and caspase-9 levels. TGP prevents testicular apoptosis, sexual dysfunction, and poor sperm motility induced by DexCS.

Unraveling the reproductive toxicity mechanisms of emerging environmental contaminants 6:2 and 8:2 diPAPs in male mice: an integrated transcriptome-metabolome investigation.

ABSTRACT Varicocele is one of the most important disorders causing infertility in men, and oxidative stress is one of the most important factors affecting testicular parenchyma damage caused by varicocele. This study explored the effect of anthocyanins on varicocele-induced testis injury in adult Wistar rats by focusing on regulating oxidative stress, Bax and Bcl-2 genes, and protein related to cell death. Rats (n = 32) were divided into four groups: Control (Sham), varicocele, varicocele + anthocyanin, and anthocyanin alone. At the end of the study (week 8), the animals were sacrificed, and H&E staining was used for testicular histopathology. The IHC method was used for the detection of Bax and Bcl-2 protein expression, and TUNEL assays were used to analyze testicular Apoptosis. Additionally, serum levels of oxidative stress markers – MDA, SOD, and GPx – were assessed by ELISA, and RT-qPCR analyzed the mRNA expression of Bax and Bcl-2. Histological analysis revealed notable improvements in Johnsen’s score, epithelial thickness, and seminiferous tubule diameter in the varicocele + anthocyanin group relative to the varicocele-only group (p < 0.005). Protein and mRNA expression of Bax significantly increased in the varicocele group (p < 0.005), while treatment with anthocyanin enhanced Bcl-2 expression (p < 0.005). Furthermore, the rate of apoptotic positive germ cells decreased when the rats received anthocyanin. Moreover, anthocyanin increased serum levels of GPx and SOD while decreasing MDA levels in the treatment group compared to rats with varicocele (p < 0.005). These outcomes suggest that anthocyanin may moderate testicular injury from varicocele, primarily through its antioxidative properties.

Purpose: To determine whether the administration of Formula X (containing Euricoma longifolia radix extract, Pimpinella pruatjan radix extract, Tribulus terrestris fructus extract, Zingiber officinale rhizome extract, and Areca catechu seed extract) can improve the quality of spermatozoa in male Wistar rats. Methods: Fourteen (14) male Wistar rats were divided evenly into 2 groups: the treatment group (given 45 mg/kg of Formula X) and normal control (given distilled water) orally for 48 days. Thereafter, the rats were sacrificed and their cauda epididymis was extracted and the motility, morphology, and total sperm count were determined. Results: The results reveal that the treatment group had a significantly higher mean sperm motility (3.71 ± 0.49) compared to control (3.0 ± 0.0; p &lt; 0.05). Although both groups had normal average morphology, the treatment group showed a significantly higher value (76.85 ± 0.48 %) in comparison with normal control (63.45 ± 1.26 %; p &lt; 0.05). In addition, the treatment group had a significantly higher mean total spermatozoa count (32.86 ± 3.93 million/mL) compared to normal control (16.43 ± 4.76 million/mL; p &lt; 0.05). Conclusion: Formula X has a spermatogenic effect, which implies improved spermatozoa quality. Studies to explore the effect of Formula X on testicular histopathology and spermatozoa viability would be required.

Non-obstructive azoospermia (NOA) is one of the most severe manifestations of male infertility, accounting for up to 70% of azoospermic cases and affecting approximately 1% of the male population. Advances in genomics and epigenetics have transformed our understanding of NOA from a primarily idiopathic condition into a biologically heterogeneous disorder driven by diverse molecular mechanisms. This review synthesizes the current knowledge of the genetic and epigenetic landscape of NOA, integrating chromosomal abnormalities, single-gene mutations, and non-coding RNA (ncRNA) dysregulation. First, we systematically examine classical and emerging chromosomal defects-including karyotype anomalies, Y-chromosome microdeletions, and structural rearrangements-that disrupt meiotic pairing and chromatin organization. Next, we explore syndromic and non-syndromic monogenic mutations affecting meiotic regulators, DNA repair factors, transcription regulators, and chromatin remodelers. Particular emphasis is placed on recently identified genes such as SYCP1, SYCE1 and HORMAD1, whose pathogenic variants are frequently linked to spermatogenic arrest. We then discuss the expanding role of ncRNAs-including microRNAs, PIWI-interacting RNAs, long non-coding RNAs, and circular RNAs-in regulating germ cell apoptosis, transposon silencing, and epigenetic reprogramming. Furthermore, we highlight the translational potential of these molecular insights (including gene variants, ncRNAs and protein) in clinical applications. Genotype-guided sperm retrieval, non-invasive biomarkers, and multi-omic approaches are discussed as promising tools to improve diagnosis and treatment. Moreover, we summarize current and emerging strategies for the treatment and fertility preservation of NOA. Finally, we identify persisting challenges, such as genotypic heterogeneity and incomplete functional validation, and emphasize the need to elucidate interactions between ncRNA and classical genetic pathways to uncover regulatory hierarchies underlying NOA. By integrating molecular genetics with testicular histopathology and clinical phenotypes, this review highlights emerging genetic and ncRNA biomarkers and underscores their potential applications in the clinical management of NOA. Ultimately, a comprehensive understanding of the genetic and epigenetic underpinnings of NOA will be essential for advancing precision diagnostics and improving reproductive outcomes in affected men.

The ameliorative effects of L-arginine on testicular and genotoxic toxicity induced by chronic exposure to cadmium chloride in male murine models.

This study explored the potential of Jasminum sambac (jasmine) and Mentha piperita (peppermint) as natural sexual stimulants (aphrodisiacs) to enhance sexual activity in males and females through the use of essential oils. The main objective was to identify the most effective combination of essential oil formulations, which were evaluated through several biological and physiological parameters, including sexual behavior in mice, sperm quality, testicular histopathology, and wax-based formulations as the delivery medium. Three formulations were tested: P1 (3:1:1), P2 (1:3:1), and P3 (1:1:3), administered to mice using a humidifier via inhalation. Sexual behavior was assessed through mounting latency and mounting frequency as indicators of libido. Sperm quality parameters included morphology, motility, viability, and sperm count. Testicular histopathology was examined to evaluate tissue structure and spermatogenesis activity. The results demonstrated that the P1 formulation was the most effective, showing the highest aphrodisiac activity. In the sexual behavior test, P1 recorded an activity value of 21±6.2, indicating enhanced libido. Sperm quality testing also confirmed the superiority of P1, with the lowest sperm abnormalities (5.3±1.53), highest motility (82±7.5), highest viability (85.6±4.58), and the greatest sperm count (53,300±16,653.3). Although testicular histopathology revealed a slight reduction in spermatogenic cells across all treatment groups, the change was not significant in the P1 group compared to the control. Overall, the P1 formulation improved libido and sperm quality more effectively than P3 and the control group, indicating its potential to be developed as an aphrodisiac preparation based on natural essential oils.

Guilu Erxian glue mitigates spermatogenesis dysfunction through HIF-1α/SLC7A11-mediated ferroptosis inhibition: An integrated metabolomics and network pharmacology study.

Abstract Reproductive efficiency in rams is influenced by seasonal variations. The aim was to investigate the effects of the breeding versus the non-breeding season on testicular morphometrics, sperm quality, and testicular histopathology in sheep. A total of 28 testicles were collected from slaughtered rams during two different seasons: namely, breeding (September to November, n=10) and non-breeding (July to August, n= 18). Testicular morphometric parameters, including testicular and epididymal weights and lengths were recorded. Epididymal spermatozoa were recovered and evaluated for their motility (by CASA), viability and abnormalities (by eosin-nigrosin), and membrane integrities (using hypo-osmotic swelling test; HOS). Sections from testicles and epididymides were also prepared for histopathological examination. Data were analyzed by student t-test and Chi-Square test. Results showed that testicular and epididymal wights and lengths were higher (P&amp;lt; 0.05) in breeding than in non-breeding seasons. Sperm motility was higher (P&amp;lt; 0.05) in the breeding than in non-breeding seasons (75.0% vs. 55.0%). Percentages of live spermatozoa and sperm with intact membrane (HOS+) were also higher (P&amp;lt; 0.05) in the samples collected during the breeding season than those retrieved in non-breeding Samples collected during breeding season had lower (P&amp;lt; 0.05) sperm abnormalities than those obtained during the non-breeding season. Histopathological analysis revealed that 100% of of the samples collected during the breeding season exhibited moderate to normal spermatogenesis, however, 22% of the samples collected during the non-breeding season showed poor or arrested spermatogenesis with reduced or absent sperm storage indicating degeneration in the seminiferous tubules of the testicles. These findings confirm that rams experience significant seasonal changes in testicular function, sperm production, and sperm quality. Testicular and epididymal growth peak during the breeding season, while spermatogenesis and sperm motility decline in the non-breeding season. The data suggest that rams exhibit seasonal reproductive adaptations, with some experiencing temporary infertility in the non-breeding months due to lower motility, decreased viability, and increased abnormalities. These results provide valuable insights for optimizing breeding management strategies in sheep production.

diabetes mellitus (DM) is associated with testicular damage, leading to male infertility. This study investigates the effects of telmisartan, moderate-intensity aerobic exercise, and their combination on testicular histopathology in a streptozotocin-induced diabetic rat model. male Wistar rats were divided into five groups: healthy control (K0), diabetic control (K1), telmisartan monotherapy (K2), aerobic exercise monotherapy (K3), and combination therapy (K4). Diabetes was induced using streptozotocin (STZ), and treatments were administered for 10 weeks. Testicular histopathology was assessed by evaluating Johnsen score, Sertoli cell count, Leydig cell count, and seminiferous tubule diameter. diabetic rats (K1) showed significant declines in Johnsen score, Sertoli and Leydig cell counts, and seminiferous tubule diameter (p<0.05). Telmisartan (K2) and combination therapy (K4) significantly improved all parameters, with values approaching those of healthy controls (K0). Aerobic exercise (K3) improved seminiferous tubule diameter but had limited effects on Johnsen score, Sertoli, and Leydig cells. Kruskal- Wallis, Mann-Whitney U, ANOVA, Games-Howell, and LSD tests confirmed these findings. Telmisartan, either as monotherapy or in combination with moderate-intensity aerobic exercise, effectively ameliorates testicular damage in diabetic rats. Aerobic exercise alone has a partial protective effect. These findings suggest potential therapeutic strategies for preventing diabetes-induced male infertility.

Diabetes mellitus (DM) has been linked to reproductive impairments. Medicinal plants have shown potential in alleviating diabetes-induced reproductive dysfunction in male. The Primary aim of the study was to assess the influence of an extract derived from Cassia fistula pod on reproductive hormone levels and testicular dysfunction in diabetic rats. A streptozotocin (STZ) dose (60 mg/kg b.wt.) was intraperitoneally (i.p.) injected to Wistar male rats to induce diabetes. 36 male rats were randomly assigned to six different groups: a healthy control group, a diabetic control group, three diabetic groups administered varying amount of Cassia fistula extract (100, 250, 500 mg/kg body weight per day), and a diabetic group receiving glibenclamide (5 mg/kg body weight per day). The treatment was given every day for 60 consecutive days. Levels of reproductive hormones including follicle-stimulating hormone (FSH), testosterone and luteinizing hormone (LH), along with oxidative stress in testicular tissue, were assessed. Histomorphometric and histopathological alterations in the testes were also examined. Diabetic control group exhibited significant decline in testicular weight, the testicular germ cells population, seminiferous tubular diameter and reproductive hormones like testosterone, FSH and LH as compared to control rats. Additionally, significant rise in lipid peroxidation (TBARS) alongside a simultaneous reduction in SOD and CAT activities as well as ascorbic acid and glutathione levels within the testicular tissues were observed compared to control rats. The administration of Cassia fistula extract or glibenclamide via oral route in diabetic rat led to improvements in serum insulin and reproductive hormone concentrations. Additionally, a reversal of histopathological and histomorphometric changes was noted relative to the diabetic reference group. Furthermore, the administration with the extract decreased lipid peroxidation and increased antioxidant levels in testicular tissue in comparison to the untreated diabetic rats. The outcomes of this research reveal that the hydroalcoholic extract from Cassia fistula pod exhibits significant antioxidant activities and can also modulate testicular dysfunction in diabetic male rats.

Background:Red beans are rich in bioactive compounds, such as flavonoids, anthocyanins, tannins, and polyphenols, which have high antioxidant activity and are believed to provide protection against testicular tissue exposure to toxic substances, including cigarette smoke.Aim:This study aims to scientifically analyze the effect of fermented red bean extract on the number of spermatogenic cells in male mice exposed to cigarette smoke.Methods:This experimental study included 25 mice. Group K (–) was administered 0.5 ml of 1% CMC-Na, group K (+) was administered 0.5 ml of 1% CMC-Na and exposed to cigarette smoke, and groups P1, P2, and P3 were administered fermented red bean extract (Phaseolus vulgaris L.) at doses of 26 mg/kgBW, 52 mg/kgBW, and 104 mg/kgBW, respectively, and exposed to cigarette smoke. Each group was given one cigarette per day for 36 days. Testicular histopathology preparations were made with hematoxylin and eosin staining and continued with spermatogenic cell counting. Data were analyzed using the one-way analysis of variance and continued with Duncan’s test (p < 0.05).Results:The results showed that the K(–) group had the highest number of spermatogenic cells, while the K(+) group had the lowest number of spermatogenic cells. The P2 group was the most effective method in maintaining the number of cells because the P2 group had a good number of cells even with a lower dose compared to the P3 group.Conclusion:This study concluded that the dose of 52 mg/kgBW has the best potential. Fermented red bean extract contains isoflavones that act as free radical scavengers by donating electrons to reactive oxygen species to stabilize the molecule.

BackgroundCadmium (Cd) is a hazardous heavy metal, and its exposure can lead to a range of health issues, including significant adverse effects on reproductive health in animals and humans. Recently, there has been increasing recognition of the antioxidant benefits of Ceratonia siliqua extract (CSE).ObjectiveTo evaluate the therapeutic potential of CSE in mitigating testicular injury and spermatogenesis impairment induced by Cd.Materials and MethodsIn this experimental study, 40 adult male BALB/c mice (8–12 wk, 30 5 gr) were randomly divided into 4 groups (n = 10/each): control, Cd (0.35 mg/kg), CSE (100 mg/kg), and CSE+Cd (100 mg/kg + 0.35 mg/kg). Adult male mice were intraperitoneally injected for one cycle of spermatogenesis (35 days). Sperm parameters, sperm DNA integrity, testicular histopathology status, testosterone hormone level, and testicular levels of malondialdehyde, nitric oxide, and total antioxidant capacity were assessed.ResultsCSE restored spermatogenesis by improving sperm count, motility, viability, morphology, and chromatin integrity (p 0.01). Testosterone levels and the histopathology of the testes also showed significant improvement in the CSE-administrated groups (p 0.001). More notably, Cd administration significantly induced oxidative stress in testicular tissue (p 0.001). Also, CSE restored antioxidant status by enhancing total antioxidant capacity levels and ameliorating nitric oxide and malondialdehyde levels (p 0.001).Conclusion Administering CSE could potentially enhance testis function and sperm parameters against chronic Cd exposure-induced reproductive toxicity, likely due to improving testosterone secretion and its antioxidant properties.

Despite extensive research on lead (Pb) toxicity's detrimental effects on male reproductive health, its precise mechanisms remain elusive, and the synergistic protective effects of resveratrol (RSV) and β-glucan (βG) on fertility parameters are underexplored. This study investigates their combined efficacy against Pb-induced reproductive toxicity in male mice. Forty 6-8-week-old mice were randomly divided into five groups: Group I (Control): received normal saline; Group II (Pb): received lead acetate (50mg/kg); Group III (Pb + RSV): received Pb and RSV (20mg/kg); Group IV (Pb + βG): received Pb and βG (50mg/kg); Group V (Pb + RSV + βG): received Pb, RSV, and βG. Mice received treatments for 35 days. Sperm parameters (count, motility, viability, DNA damage), oxidative stress markers (TAC, SOD, GPx, MDA), hormone levels (testosterone, LH, FSH), testicular histopathology, and apoptosis-related gene expression (Bax, Bcl-2, caspase-3) were evaluated. The Pb + RSV + βG group, compared to the Pb-only group, showed a 61.5% increase in sperm motility, 68.1% reduction in sperm DNA damage, and 87.2% increase in sperm count. It also exhibited a 63.9% reduction in MDA levels and increased TAC, SOD, and GPx levels. Hormone levels and anti-apoptotic Bcl-2 expression increased, while Bax and caspase-3 decreased significantly. These preclinical findings suggest that RSV and βG may mitigate Pb-induced reproductive toxicity in mice via antioxidant and anti-apoptotic mechanisms.

BackgroundObesity is associated with hormonal imbalance, increased oxidative stress, and inflammation in the testis. These conditions adversely affect sperm quality, leading to impaired male fertility. Therefore, therapeutic interventions to counteract the adverse effects of obesity are crucial. This study explored the therapeutic effects of 4-hydroxyisoleucine (4-HIL) on fertility in male mice with diet-induced obesity.MethodsC57BL6 male mice (n=45) were randomly divided into normal diet (ND, n=15) and high-fat diet (HFD, n=30) groups for 10 weeks. The HFD group was then randomized into untreated (HFD, n=15) and 4-HIL-treated (200 mg/kg/day, intraperitoneal injection, HFD + 4-HIL group, n=15) for 6 weeks. ND and HFD controls received saline (0.3 mL/30 g body weight) throughout the intervention period. Comprehensive evaluations included (1) metabolic assessments (body weight, glucose, insulin and pyruvate tolerance tests, and blood serum lipids), (2) sperm analysis (count, concentration, and morphology), (3) fertility testing (mating trials and in vitro fertilization), (4) testicular histopathology (fat deposition and apoptosis), (5) biochemical assays (reproductive hormones, oxidative stress markers, and inflammatory cytokines), and (6) molecular analyses (mRNA sequencing and qPCR validation of differentially expressed genes).Results4-HIL treatment improved metabolic parameters, including reduced weight gain, enhanced glucose tolerance, and optimized blood serum lipids, compared to HFD controls. Treated mice exhibited superior sperm quality with increased count and concentration, reduced histomorphological abnormalities in the testis, and attenuated oxidative stress and inflammation. Furthermore, the key spermatogenic gene expressions, including spem1 and spata24, were significantly optimized in the testes of mice treated with 4-HIL compared to those of untreated mice (HFD group).ConclusionThis study demonstrates that 4-HIL therapy ameliorates obesity-induced testicular dysfunction and improves fertility markers in mice. The beneficial effects of this compound on metabolic parameters, sperm quality, and spermatogenic gene expression suggest its potential as a therapeutic agent for obesity-related male infertility. Further research is warranted to elucidate the underlying mechanisms and assess the clinical translatability of these findings.

Ionizing radiation is a well-known environmental stressor capable of generating excessive reactive oxygen species (ROS), leading to oxidative damage in sensitive tissues, including the reproductive system. While oxidative stress is increasingly implicated in male reproductive dysfunction, the long-term effects of low-dose-rate (LDR) radiation on testicular structure and oxidative status remain underexplored. In this study, mice were exposed to continuous LDR radiation (0.39, 1.29, and 3.46 mGy/h) for 21 days to assess testicular histopathology and oxidative status. Although testis weight did not significantly differ among groups, histological analysis revealed basal membrane disruption and reduced spermatogenic cell populations in irradiated groups. Masson’s Trichrome and Sirius Red staining demonstrated dose-dependent collagen deposition, indicating progressive testicular fibrosis. TUNEL assays confirmed increased germ cell apoptosis in the mid- and high-dose-rate groups. ROS levels were significantly elevated only in the highest-dose group, suggesting a threshold-dependent oxidative stress response. These findings indicate that chronic LDR radiation induces testicular damage primarily through apoptosis and fibrosis, with oxidative stress potentially contributing at higher exposure levels.

Topiramate (TPM), a widely used anticonvulsant, has been documented to induce testicular dysfunction through its pro-oxidant properties, leading to cellular damage and hormonal abnormalities in the testes. This damage is characterized by increased malondialdehyde (MDA) levels and decreased activity of antioxidant enzymes such as catalase (CAT) and superoxide dismutase (SOD), which are essential for reducing oxidative stress. This study aimed to evaluate the effects of TPM and Gallic Acid (GA) on reproductive health in male rats. Forty mature Sprague-Dawley rats, aged 16 to 18 weeks and weighing between 180 and 200 g, were divided into four experimental groups (10 rats each): a control group, a TPM-treated group, a TPM + GA-treated group, and a GA-only group. The rats received TPM (18 mg/kg) orally for 60 days, with or without GA (50 mg/kg) administered orally for the same period. Testicular tissues were examined for oxidative stress markers (MDA, SOD, CAT), sperm motility, hormonal concentrations (Testosterone, gonadotropin-releasing hormone [GnRH], and 17β-hydroxysteroid dehydrogenase type 3 [17β-HSD3]), and histological changes. The results showed that TPM significantly increased MDA levels while decreasing CAT and SOD activity, indicating oxidative stress compared to the control group. Sperm motility was also impaired in the TPM-treated group. However, GA treatment led to a notable reduction in MDA levels and restored antioxidant enzyme activity toward normal levels. Hormonal analysis revealed that TPM affected testosterone and GnRH levels, although GA partially mitigated these changes. Immunohistochemical and histological assessments demonstrated considerable testicular damage in the TPM group, whereas the GA-treated group showed slight improvements in testicular histopathology and reduced cellular death. In conclusion, GA (50 mg/kg) exhibited a protective effect against TPM-induced testicular dysfunction.

The Effects of Boron on Sperm Qualities and Testicular Histopathology in Animal Studies: A Systematic Review

Restorative mechanisms of Shugan Yiyang capsule on male infertility through 'pharmaco-metabo-net' tripartite correlation analysis.

Multi-mechanistic effects of bisphenol A on testicular dysfunction and endocrine disruption in adult male Labeo bata: oxidative stress, inflammation, and dysregulated energy sensors.