Number Of Spermatogenic Cells Research Articles

White guava (Psidium guajava L.) fruit juice ameliorated the number of spermatogenic cells in rats (Rattus Novergicus) exposed to lead acetate

White guava (Psidium guajava L.) fruit juice ameliorated the number of spermatogenic cells in rats (Rattus Novergicus) exposed to lead acetate

Filtered and Unfiltered Cigarette Effect on White Rats Male Reproduction (Rattus norvegicus L.)

Cigarette smoke can cause various health problems, one of which is a decrease in fertility in males. The purpose of this study was to determine the effect of smoke exposure between with and without filter hand-rolled kretek cigarettes (SKT) and white cigarettes on white rats male reproduction: sperm quality in the form of sperm motility and viability, also the structure of seminiferous tubules. This study used 5 treatment groups (KP), namely controls not given exposure to cigarette smoke (KP0), exposure to unfiltered kretek cigarette smoke (KP1), exposure to filtered kretek cigarettes (KP2), exposure to unfiltered white cigarette smoke (KP3) and exposure to secondhand smoke. filtered white cigarettes (KP4) with treatment placement arrangement in a Completely Randomized Design (CRD) and if there is a significant difference, a Duncan Multiple Range Test (DMRT) is carried out. The results obtained on sperm quality, namely sperm motility and viability, decreased. Good quality at KP0 then the quality decreases successively starting from KP1, KP2, KP3 and KP4. The results on histpathological observations of the seminiferous tubules, there were no pathological changes found in all treatments. In contrast to the histopathology of seminiferous tubules, the number of spermatogenic cells showed a significant difference between treatments with the least average spermatogenic count and a decrease in the number of cells from spermatogonia cells to spermatid cells, namely in the treatment group exposure to filtered white cigarette smoke (KP4). Overall, the exposure that caused the most severe impact on all measured parameters was in the treatment group of filtered white cigarette smoke (KP4).

RNA sequencing profiles reveals progressively reduced spermatogenesis with progression in adult cryptorchidism.

The fertility of cryptorchidism patients who didn't perform corrective surgery will decrease with age. Herein, we elucidate the histological alterations and underlying molecular mechanism in patients with an increase in the disease duration from 20 to 40 years. Testicular tissues were obtained from three patients with cryptorchidism, ranging in age from 22 to 44 years. Three benign paracancerous testicular samples of matched ages were used as controls. The normal and undescended testicular tissues were stained with hematoxylin and eosin (HE) and immunofluorescence and all six testicular samples were subjected to RNA sequencing. RNA sequencing data were subjected to gene set enrichment analysis (GSEA), Kyoto Encyclopedia of Genes and Genomes (KEGG), protein-protein interaction (PPI) network analysis, and Gene Ontology (GO) searches. Real-time reverse transcriptase polymerase chain reaction was used to confirm the DEGs. The seminiferous tubules' basement membrane thickens with age in healthy testes. As the period of cryptorchidism in the cryptorchid testis extended, the seminiferous tubules significantly atrophy, the number of spermatogenic cells declines, and the amount of interstitial fibrous tissue increases in comparison to normal tissues. The number of germ cells per cross-section of seminiferous tubules was significantly lower in cryptorchidism than in normal testicular tissues, according to immunofluorescence staining, but the number of Sertoli cells remained stable. RNA sequencing analysis identified 1150 differentially expressed genes (DEGs) between cryptorchidism and normal testicular tissues (fold change >2 and p<0.05), of which 61 genes were noticeably upregulated and 1089 were significantly downregulated. These genes were predominantly linked to sperm development and differentiation, and fertilization, according to GO analysis. Meiosis pathways were significantly downregulated in cryptorchidism, according to KEGG pathway analysis and GSEA (P<0.001). PPI analysis was used to identify the top seven downregulated hub genes (PLCZ1, AKAP4, IZUMO1, SPAG6, CAPZA3, and ROPN1L), which were then further verified by qPCR. By describing the histological changes and differential gene expression patterns in adult cryptorchid patients of different age groups, we discovered the progression mechanisms of undescended testes in adults with aging and identified seven significantly downregulated hub genes (PLCZ1, AKAP4, IZUMO1, SPAG6, CAPZA3, and ROPN1L) in cryptorchid testis compared to normal testicular tissues. These genes played a role in the process of spermgenesis and are directly linked to the steady decline in fertility caused by cryptorchidism. Our study provided a better understanding of the molecular mechanisms underlying the loss of spermatogenesis in adult cryptorchidism, and give support for the development of adult cryptorchidism treatments.

The Effect of Kepel Fruit Extract on The Number of Spermatogenic Cells in Mice (Mus musculus)

The Kepel plant (Stelechocarpus burahol) has garnered research interest due to its diverse secondary metabolites like alkaloids, flavonoids, polyphenols, saponins, triterpenoids, and quinones, thought to impact fertility. This research aims to determine the effect of kepel fruit extract on the number of spermatogenic cells in mice (Mus musculus), consisting of spermatogonia cells, spermatocyte cells, and spermatid cells. Employing a Posttest-Only Control Design, Kepel extraction used 95% ethanol via maceration. 25 mice were divided into groups: positive control, negative control, groups I, II, and III exposed to 50, 100, and 150 mg/kgBW Kepel extract for 36 days. Randomized group design determined sample selection, assessing seminiferous tubules via four fields of vision. Statistical analysis involved one-way ANOVA (5% significance) and LSD test for disparities. Data were analyzed using SPSS 25. ANOVA revealed no significant difference (p &gt; 0.05) in mean spermatogonia and spermatocyte cell counts between control and treatment groups (I, II, III). However, groups I, II, and III exhibited significant spermatid cell reduction. In conclusion, Kepel fruit extract at 50 mg/kgBW, 100 mg/kgBW, and 150 mg/kgBW could lower spermatid cell count, indicating potential as an antifertility agent.

The effects of coadministration of curcumin and vitamin E on the reproductive system of diabetic male rats; An experimental study

IntroductionDiabetes causes testicular damage due to oxidative stress. Nowadays, the use of vitamins and antioxidants is one of the common methods to treat this disease. Therefore, the aim of this study is to investigate the effect of single and combined administration of these two substances on the reproductive system of male diabetic rats. Method and materialsIn this study, 48 adult male Wistar rats weighing 250–270 grammes were divided into 6 groups: Control group, diabetic group, vehicle group, vitamin E, curcumin group, and vitamin E and curcumin group. The control group was the healthy group, and in the other groups, the rats were made diabetic by streptozotocin (60 mg/kg/ip). The vehicle group received 1 ml of olive oil, the vitamin E group (100 mg/kg/ip) received Vit.E, and the curcumin group (50 mg/kg/ip) received Cu. The group of rats received vitamin E and curcumin. At the end of the sixth week after treatment, blood was taken from the rats and biochemical analysis was performed to check the amount of malondialdehyde (MDA), LH hormones and serum testosterone, then the rats were killed and their testes and epididymides were removed. The weight of the testes and sperm parameters, the maturity of sperm nuclei and the integrity of their DNA were checked. The number of spermatogenic cells was determined by histological examination. ResultsThis study showed that diabetes caused a decrease in testicular weight, sperm count, motility, and viability, an increased percentage of sperm with immature nuclei, and an increased percentage of sperm with denatured DNA. In addition, diabetes decreased the average number of matogenic sperm, and biochemical results showed that diabetes increased the level of MDA and decreased the level of the hormones LH and testosterone. Treatment with vitamin E, curcumin and their combination improved all these parameters, and this improvement was significant in the Toam group. ConclusionCombined administration of vitamin E and curcumin in diabetic rats significantly improves sperm parameters, matogenic sperm count, and improves MDA levels, LH, and serum testosterone compared with separate treatment.

Effect of ethanolic extract of Moringa oleifera leaves on the number of spermatogenic cells and Leydig cells of gentamicin-induced rats

This study aimed to determine the effect of the administration of ethanolic extract of Moringa oleifera leaves on the number of spermatogenic and Leydig cells of gentamicin-induced rats (Rattus norvegicus). This study used 25 white male rats divided randomly into five groups. The rats were injected with Gentamicin 5 mg/kg bw/day subcutaneously for 14 days, except for the negative control group (NCG). During the next 14 days, the rats were given oral moringa leaf extract of 200, 316, and 500 mg/kg bw/day respectively for E200, E316, and E500 groups. The data were analyzed with one-way ANOVA followed by Duncan's test. The results showed that the number of spermatogonia, spermatocytes, and spermatids in NCG was higher (p &lt;0.05) than in the other groups. The number of spermatogonia, spermatocytes, and spermatids in the E316 group was higher (p &lt;0.05) than in PCG. The number of spermatogonia in the E500 group was higher (p &lt;0.05) than in the E316 group, but the number of spermatocytes and spermatids in the E316 group was similar (p &gt;0.05) to the E500 group. The number of PCG Leydig cells was the smallest (p &lt;0.05) compared to the other groups. The administration of Moringa leaves extract in the E200 and E316 groups increased (p &lt;0.05) the number of Leydig cells compared to PCG. The number of Leydig PCG cells in the E500 group was not significantly different (p &gt;0.05) from that of the NCG. It could be concluded that administration of Moringa oleifera leaves extract restored the number of spermatogenic and Leydig cells of gentamicin-induced rats.

Effect of ethanolic extract of Moringa oleifera leaves on the number of spermatogenic cells and Leydig cells of gentamicin-induced rats

This study aimed to determine the effect of the administration of ethanolic extract of Moringa oleifera leaves on the number of spermatogenic and Leydig cells of gentamicin-induced rats (Rattus norvegicus). This study used 25 white male rats divided randomly into five groups. The rats were injected with Gentamicin 5 mg/kg bw/day subcutaneously for 14 days, except for the negative control group (NCG). During the next 14 days, the rats were given oral moringa leaf extract of 200, 316, and 500 mg/kg bw/day respectively for E200, E316, and E500 groups. The data were analyzed with one-way ANOVA followed by Duncan's test. The results showed that the number of spermatogonia, spermatocytes, and spermatids in NCG was higher (p &lt;0.05) than in the other groups. The number of spermatogonia, spermatocytes, and spermatids in the E316 group was higher (p &lt;0.05) than in PCG. The number of spermatogonia in the E500 group was higher (p &lt;0.05) than in the E316 group, but the number of spermatocytes and spermatids in the E316 group was similar (p &gt;0.05) to the E500 group. The number of PCG Leydig cells was the smallest (p &lt;0.05) compared to the other groups. The administration of Moringa leaves extract in the E200 and E316 groups increased (p &lt;0.05) the number of Leydig cells compared to PCG. The number of Leydig PCG cells in the E500 group was not significantly different (p &gt;0.05) from that of the NCG. It could be concluded that administration of Moringa oleifera leaves extract restored the number of spermatogenic and Leydig cells of gentamicin-induced rats.

Dibutyl Phthalate Adsorbed on Multiwalled Carbon Nanotubes Causes Fetal Developmental Toxicity in Balb/C Mice.

This study investigated whether using multiwalled carbon nanotubes (MWCNTs) as a carrier for dibutyl phthalate (DBP) could delay the degradation rate of DBP in mice and increase its estrogen-like interference effect. Pregnant Balb/C mice were divided into four groups and exposed to different treatments via tail-vein injection every 3 days until gestational day 20. The female and male mice were then sacrificed for toxicological study. The results showed that the combination of MWCNTs and DBP resulted in a higher fetal mortality rate than if the mice were exposed to MWCNTs or DBP alone. H&E staining showed that the estrous period of the exposed mice was delayed, the development of oocytes was blocked in the combination group, the number of spermatogenic cells decreased, and the quality of sperm decreased. Our experiment showed that the expression levels of the genes involved in sex hormone synthesis in the testis and ovaries were significantly increased after combined treatment compared with the MWCNT group (p < 0.01). The study suggests that DBP degradation is delayed when absorbed on MWCNTs, which increases its estrogen-like interference and interferes with fetal development, ultimately leading to increased fetal mortality.

Icariin Ameliorates Spermatogenesis Disorder in Obese Mice Induced by High-Fat Diet through Regulating the Glycolytic Pathway.

Obesity is a global threat for male infertility, which can cause spermatogenic dysfunction. However, there are no available drugs for the treatment of obesity-induced spermatogenesis dysfunction. This study characterizes the protective effects of icariin (ICA) on spermatogenesis dysfunction in obese mice. Obese mice are induced by a high-fat diet to determine whether ICA has a protective effect. ICA treatment reduces body weight and the proportion of abnormal sperm, brings about a recovery of sperm count, and the number of spermatogenic cells. ICA treatment improves histopathological changes of the testes and inhibits testicular apoptosis, as evidenced by reduced the expression of Bax and increased the expression of Bcl-2, PCNA, WT1, GATA-4, vimentin, HK2, PKM2, and LDHA in the testes. In vitro, TM4 cells are treated with 0.4mm palmitic acid (PA) to induce Sertoli cell injury, and are then utilized for ICA treatment. ICA improves PA-induced decreased TM4 cells viability, reduces the levels of lactate, and increases the levels of pyruvate and the expression of HK2, PKM2, and LDHA and restores the glycolytic process in vitro. ICA ameliorates spermatogenic dysfunction in obese mice by regulating glycolytic activity, providing effective strategies for obesity treatment.

Toxic effects of Tripterygium glycoside tablets on the reproductive system of male rats by metabolomics, cytotoxicity, and molecular docking

BackgroundTripterygium glycoside tablets (TGT) is the most common preparation from Tripterygium wilfordii Hook F, which is widely used in clinical for treating rheumatoid arthritis (RA) and other autoimmune diseases. However, its serious reproductive toxicity limits its application. PurposeThis study aimed to elucidate the toxic effects of TGT on the reproductive system of male RA rats and its potential toxic components and mechanism. MethodsCollagen-induced arthritis (CIA) rat model was established, and TGT suspension was given at low, medium, and high doses. Gonadal index, pathological changes, and the number of spermatogenic cells were used to evaluate the toxic effects of TGT on the reproductive system. Non-targeted metabolomics of testicular tissue was conducted by UHPLC-QTOF/MS. Combined with network toxicology, the key targets of TGT-induced reproductive toxicity were screened and RT-qPCR was used to validation. In vitro toxicity of 19 components of TGT was evaluated using TM3 and TM4 cell lines. Molecular docking was used to predict the interaction between toxic components and key targets. ResultsTGT reduced testicular and epididymis weight. Pathology analysis showed a lot of deformed and atrophic spermatogenic tubules. The number of spermatogenic cells decreased significantly (P<0.0001). A total of 58 different metabolites including platelet-activating factor (PAF), lysophosphatidylcholine (Lyso PC), phosphatidylinositol (PI), glutathione (GSH), and adenosine monophosphate (AMP) were identified by testicular metabolomics. Glycerophospholipid metabolism, ether lipid metabolism, and glutathione metabolism were key pathways responsible for the reproductive toxicity of TGT. Ten key reproductive toxicity targets were screened by network toxicology. The cytotoxicity test showed that triptolide, triptonide, celastrol, and demethylzeylasteral could significantly reduce the viability of TM3 and TM4 cells. Alkaloids had no apparent toxic effects. Molecular docking showed that the four toxic components had a good affinity with 10 key targets. All binding energies were less than -7 kcal/mol. The RT-qPCR results showed the Cyp19a1 level was significantly up-regulated. Pik3ca and Pik3cg levels were significantly down-regulated. ConclusionThrough testicular metabolomics, we found that TGT may cause reproductive toxicity through CYP19A1, PIK3CA, and PIK3CG three target, which was preliminarily revealed. This study laid the foundation for elucidating the toxicity mechanism of TGT and evaluating its safety and quality.

Protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse spermatogenesis and sperm chromatin condensation following unilateral testicular torsion

Testicular torsion is considered a urological disorder that requires immediate detorsion surgery. Ischemia/reperfusion (I/R) injury after testicular torsion detorsion causes of drastic impairment of spermatogenesis and infertility. Cell-free-based approaches seem to be a promising strategy to prevent I/R injury, they have more stable biological properties, and they contain paracrine factors of mesenchymal stem cells. The purpose of this study was to evaluate the protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse sperm chromatin condensation and spermatogenesis improvement after I/R injury. hAMSCs were isolated and characterized by RT- PCR and flow cytometry, preparation of hAMSCs secreted factors was performed. Forty male mice were randomly divided into 4 groups: sham-operated, torsion detorsion, torsion detorsion+ intratesticular injection of DMEM/F-12, and torsion detorsion+ intratesticular injection of hAMSCs secreted factors. After one cycle of spermatogenesis, the mean number of germ cells, Sertoli, Leydig, myoid as well as tubular parameters, Johnson score, and spermatogenesis indexes were evaluated by H& E and PAS stainings. Sperm chromatin condensation and relative expression of c-kit and prm 1 genes were assessed by aniline blue staining and real-time PCR, respectively. The mean number of spermatogenic cells, Leydig, myoid, Sertoli, spermatogenesis parameters, Johnson score, as well as germinal epithelial height and diameters of seminiferous tubules decreased significantly after I/R injury. The thickness of basement membrane and percentage of sperm with excessive histone significantly increased, while the relative expression of c-kit and prm 1 significantly decreased in torsion detorsion group (p 0.001). hAMSCs secreted factors remarkably restored normal sperm chromatin condensation, spermatogenesis parameters and histomorphometric organization of seminiferous tubules via intratesticular injection (p 0.001). Thus, hAMSCs secreted factors may potentially salvage torsion-detorsion-induced infertility.

Evaluation of the Protective Effect of Human Platelet-Rich Plasma on Spermatogenesis and Sperm Chromatin Condensation Following Ischemia-Reperfusion Injury Due to Spermatic Cord Torsion in Mice

Introduction: Torsion of spermatic cord induces ischemic-reperfusion injury which leads to testicular dysfunction, apoptosis of germ cells, ROS generation and infertility. Platelet-rich plasma (PRP) is a rich source of growth factors and anti-inflammatory cytokines. In this study, the effects of PRP on spermatogenesis and sperm chromatin condensation after ischemia-reperfusion injury were evaluated.&#x0D; Methods: Forty adult male mice were procured in the study. Control group, torsion/ detorsion group, negative control group: torsion/ detorsion surgery with intra testicular injection of culture medium, treatment group: torsion/ detorsion surgery with intra testicular injection of PRP. After one cycle of spermatogenesis, histomorphometric evaluation was performed by PAS staining and sperm chromatin condensation was assessed by aniline blue staining. Data was analyzed by using SPSS Statistics and ANOVA followed by Tukey test.&#x0D; Results: The mean number of spermatogenic cells, Sertoli, Leydig, Myoid, Johnson score and mean diameter and thickness of germinal epithelium were significantly decreased following ischemia-reperfusion injury. Mean thickness of seminiferous tubules basement membrane as well as sperm percentage with residual histone increased after ischemia-reperfusion injury (P &lt;0.001). After PRP treatment, there was a significant improvement in the histological structure of the tubules, cell viability, and sperm chromatin condensation (P &lt;0.001). While, a significant difference was observed in comparison with control group.&#x0D; Conclusion: PRP can partially ameliorate the complications of ischemia-reperfusion injury through its growth factors and anti-inflammatory cytokines involved in angiogenesis and antioxidant activity.

Effects of growth hormone on lipid metabolism and sexual development in pubertal obese male rats.

To investigate the effects of growth hormone (GH) on pubertal obese male rats, a rat model of high-fat diet-induced obesity was established in juvenile male rats. The model rats were divided into the treatment group (GH) and the non-treatment group (physiological saline). After 4 weeks, we measured the levels of alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), estrogen (E2), testosterone (T), and insulin-like growth factor (IGF-1). The morphological changes of the liver and testis were assessed, and the expression of aromatase was detected. The levels of ALT, AST, TC, TG, LDL-C, E2, and IGF-1 in the treatment group were significantly lower than in the non-treated model rats (P < 0.001). The levels of HDL-C and T of GH-treated rats were significantly higher than those of the non-treatment group (P < 0.001). Compared with non-treated model rats, GH-treated model rats showed reduced liver steatosis, improved morphological structure of the testicular seminiferous tubules, and an increased number of spermatogenic cells. The treatment group also showed lower expression of aromatase in the liver and testis compared with the non-treatment group. GH partially protected pubertal male rats from obesity-induced lipid metabolic disorder and sexual retardation.

The adiponectin receptor agonist, AdipoRon, promotes reproductive hormone secretion and gonadal development via the hypothalamic-pituitary-gonadal axis in chickens

Adiponectin is a key hormone secreted by fat tissues that has multiple biological functions, including regulating the energy balance and reproductive system by binding to its receptors AdipoR1 and AdipoR2. This study investigated the correlation between the levels of adiponectin and reproductive hormones in the hypothalamic-pituitary-ovarian (HPO) axis of laying hens at 4 different developmental stages (15, 20, 30, and 68 wk) and explored the effects of AdipoRon (an activator of adiponectin receptors) on the hypothalamic-pituitary-gonadal (HPG) axis and follicle and testicular Leydig cells in vitro and in vivo. The results demonstrated that the adiponectin level was significantly correlated with that of reproductive hormones in the HPO axis (e.g., GnRH, FSH, LH, and E2) in laying hens at 4 different ages. Moreover, AdipoRon could promote the expression of AdipoR1 and AdipoR2 and the secretion of reproductive hormones in the HPG axis, including GnRH, FSH, LH, P4, and T. AdipoRon could also upregulate the expression of genes related to follicular steroidogenesis (STAR, CYP19A1, CYP17A1, and CYP11A1), hepatic lipid synthesis (OVR, MTP), follicular lipid uptake (PPAR-g), and follicular angiogenesis (VEGFA1, VEGFA2, VEGFR1, ANGPT1, ANGPT2, TEK) in the oviposition period, and all of these findings were consistent with the results obtained from in vitro experiments after the transfection of small white follicles (SWFs) with AdipoRon. Furthermore, the results suggest that AdipoRon increases the diameter of testicular seminiferous tubules, the number of spermatogenic cells and sperm production in vivo and enhances the expression of AdipoR1, AdipoR2 and steroid hormones in vitro. Collectively, the findings suggest that AdipoRon could facilitate the expression and secretion of reproductive hormones in the HPG axis by activating its receptors and then improve the growth and development of follicles and testes in chickens.

Beneficial effects of Spirulina platensis on mice testis damaged by silver nanoparticles.

Silver nanoparticles (AgNPs) have been used widely in medical applications and various industries. Humans could be exposed to the risk of AgNPs toxicity through different routes. The current study aimed to investigate the role of Spirulina platensis (SP) against the side effects of AgNPs on mice testis. Adult male NMRI mice were divided into four groups: control group, SP group (300 mg/kg bwt), AgNPs (20 nm) group (500 mg/kg bwt), Co-treated group (SP + AgNPs). The groups were treated orally for 35 days. Subsequently, epididymal sperm parameters, sperm DNA integrity, daily sperm production (DSP), sexual hormones level, malondialdehyde (MDA), total antioxidant capacity (TAC) and spermatogenesis indices were measured. In addition, the histopathology of testes was evaluated using tissue processing, haematoxylin-eosin staining and stereology techniques. A significant decrease in the number of spermatogenic cells, Leydig cells and sperm parameters was observed in the AgNPs treated group. Serum levels of testosterone and TAC were decreased significantly following AgNPs treatment. Also, MDA incremented in the serum of AgNPs treated mice. The stereological analysis revealed that AgNPs exposure induced histopathological changes in the seminiferous tubules, degeneration and dissociation of spermatogenic cells. In contrast, SP co-administration significantly counteracted AgNPs reproductive toxicity impacts. SP co-exposure caused an increase in spermatogenesis indices, TAC and also a decrease in MDA. SP improved the histopathological changes of testes tissue and spermatozoa abnormalities. In parallel, SP modulated levels of testosterone, FSH and LH. Spirulina platensis exhibited the protective potential by regulating oxidative stress against AgNPs-induced reproductive toxicity. SP could be a candidate therapy against AgNPs reprotoxic impacts.

Improving the process of spermatogenesis in azoospermic mice using spermatogonial stem cells co-cultured with epididymosomes in three-dimensional culture system

AimsThis study aimed to explore the effect of epididymosomes on the proliferative efficiency of spermatogonial stem cells (SSCs) in vitro and the resumption of spermatogenesis in the azoospermic mice. Main methodsThe epididymosomes were extracted from the epididymis and characterized. SSCs were cultured in 2D (two-dimensional) and hydrogel-based 3D culture in the presence of 20 μg/mL epididymosome or 10 ng/mL GDNF. After two weeks of culture, the proliferation and purity of the separated SSCs were evaluated using the MTT test and flow cytometry, respectively. qRT-PCR was used to analyze PLZF, caspase-3, TGF-β, miR-10b, and miR-21 expression levels. Then, SSCs grown in the 3D culture system were labeled by DiI and transplanted into azoospermic mice via the efferent duct. After 2 weeks, tracing of DiI and cell homing were evaluated. Subsequently, histomorphometric studies and immunohistochemistry analysis were performed in testes after eight weeks of transplantation. Key findingsThe expression of PLZF, TGF-β, miR-10b, and miR-21 increased significantly (*p < 0.05) in the 3D + GDNF and 3D + epididymosomes groups than in the 2D group. Transplanted SSCs migrated into the seminiferous tubules of recipient mice and the number of spermatogenic cells and protein expression of PLZF, SCP3 and ACRBP in the 3D + GDNF and 3D + epididymosomes groups were considerably higher (∗ ∗ ∗ p < 0.001) compared to the azoospermic group. SignificanceThis finding indicates that culturing SSCs on decellularized testicular matrix (DTM) hydrogel with 10 ng/mL GDNF or 20 μg/mL epididymosomes could lead to an increase in SSCs proliferation which provides a sufficient number of SSCs for successful transplantation in azoospermic mice.

Effect and mechanism of Dahuang Zhechong Pills against testicular aging in rats by inhibiting necroptosis signaling pathway

To explore the effect and mechanism of Dahuang Zhechong Pills(DHZCP), a classical prescription, in improving testicular aging(TA) in vivo, the authors randomly divided 24 male rats into four groups: the normal, model, DHZCP and vitamin E(VE) groups. The TA rat model was established by continuous gavage of D-galactose(D-gal). During the experiment, the rats in the DHZCP and VE groups were given DHZCP suspension and VE suspension, respectively by gavage, while those in the normal and model groups were gavaged saline separately every day. After the co-administration of D-gal and various drugs for 60 days, all rats were sacrificed, and their blood and testis were collected. Further, various indexes related to TA and necroptosis of testicular cells in the model rats were examined and investigated, which included the aging phenotype, total testicular weight, testicular index, histopathological features of testis, number of spermatogenic cells, sex hormone level, expression characteristics of reactive oxygen species(ROS) in testis, expression levels and characteristics of cyclins in testis, and protein expression levels of the key molecules in receptor-interacting serine/threonine-protein kinase 1(RIPK1)/receptor-interacting serine/threonine-protein kinase 3(RIPK3)/mixed lineage kinase domain like pseudokinase(MLKL) signaling pathway in each group. The results showed that, for the TA model rats, both DHZCP and VE improved their aging phenotype, total testicular weight, testicular index, pathological features of testis, number of spermatogenic cells, serum testosterone and follicle stimulating hormone levels, expression characteristics of ROS and protein expression levels and characteristics of P21 and P53 in testis. In addition, DHZCP and VE improved the protein expression levels of the key molecules in RIPK1/RIPK3/MLKL signaling pathway in testis of the model rats. Specifically, DHZCP was better than VE in the improvement of RIPK3. In conclusion, in this study, the authors found that DHZCP, similar to VE, ameliorated D-gal-induced TA in model rats in vivo, and its mechanism was related to reducing necroptosis of testicular cells by inhibiting the activation of RIPK1/RIPK3/MLKL signaling pathway. This study provided preliminary pharmacological evidence for the development and application of classical prescriptions in the field of men's health.

Nicotinamide mononucleotide improves spermatogenic function in streptozotocin-induced diabetic mice via modulating the glycolysis pathway.

Spermatogenic dysfunction is one of the major secondary complications of diabetes; however, the underlying mechanisms remain ill-defined, and there is no available drug or strategy for the radical treatment of diabetic spermatogenic dysfunction. Therefore, the objective of this study is to investigate the protective effects of nicotinamide mononucleotide (NMN) on testicular spermatogenic function in streptozotocin (STZ)-induced diabetic mice. The results show that oral administration of NMN significantly increases the body and testis weight and the number of sperms. Moreover, the abnormal sperm count and the rate of sperm malformation are significantly decreased compared with the saline-treated diabetic mice. Histological analysis reveals that NMN treatment significantly increases the area and diameter of seminiferous tubules, accompanied by an increased number of spermatogenic cells and sperms. Immunohistochemistry and qRT-PCR results show that NMN increases Bcl-2 expression and decreases Bax expression in the testis. NMN also increases the protein expression of Vimentin and the mRNA expressions of WT1 and GATA4. In addition, qRT-PCR, western blot analysis and immunohistochemistry results also show that NMN increases the expressions of glycolysis-related rate-limiting enzymes including HK2, PKM2, and LDHA. In summary, this study demonstrates the protective effects of NMN on the testis in an STZ-induced diabetic mice model. NMN exerts its protective effects via reducing spermatogenic cell apoptosis by regulating glycolysis of Sertoli cells in diabetic mice. This study provides an experimental basis for the future clinical application of NMN in diabetes-induced spermatogenic dysfunction.

Dendrobium nobile-derived polysaccharides ameliorate spermatogenic disorders in mice with streptozotocin-induced diabetes through regulation of the glycolytic pathway

Dysfunction of spermatogenesis is a major complication of diabetes mellitus (DM). This study characterized the protective effects of Dendrobium nobile-derived polysaccharides (DNP) against spermatogenetic dysfunction in mice with streptozotocin (STZ)-induced diabetes. The diabetic mice had lower body and testicular mass, and fewer spermatozoa with a higher incidence of malformation. The testicular histology showed disordered narrow seminiferous tubules covering a smaller area, and fewer spermatogenic cells. Moreover, the qRT-PCR analysis indicated that DM was associated with high expression of the pro-apoptotic factor Bax and low expression of the anti-apoptotic factor Bcl-2 in the testes. The qRT-PCR and immunohistochemical analysis clarified that DM was also associated with low testicular expression of the Sertoli cell (SC) markers GATA-4, WT1, and vimentin, and genes encoding the glycolytic rate-limiting enzymes LDHA, PKM2, and HK2. DNP treatment increased the body and testicular masses, sperm count, and number of spermatogenic cells of the mice, and reduced the proportion of abnormal sperm. DNP also reduced the expression of Bax, and increased that of Bcl-2, GATA-4, WT1, vimentin, LDHA, PKM2, and HK2, in the testes of the diabetic mice. Thus, DNP protects against spermatogenic dysfunction in diabetic mice by inhibiting apoptosis and activating the glycolytic pathway in their testes.

Protective effect of indole-3-carbinol on testis of a high fat diet induced obesity

ABSTRACT We investigated the effects of obesity caused by a high fat diet (HFD) on rat testes and evaluated the possible protective effects of indole-3-carbinol (IND). We used 24 8–10-week-old 200 g male rats randomly assigned to 4 groups: non-obese control (NC), obese control (OC), non-obese IND group (NI), obese + IND group (OI). Testis samples were examined using stereological, immunohistochemical, biochemical and histological methods. The number of spermatogenic cells, Leydig cells, mean volume of testes and seminiferous tubules was significantly decreased in the OC group compared to the NC group, but these values were increased significantly in the OI group compared to the OC group. We found a significant increase in catalase and myeloperoxidase activities in the OC group compared to the NC group. In the OI group, catalase and myeloperoxidase levels were decreased compared to the OC group. TUNEL-positive cells also were increased in the OC group compared to the NC group (p < 0.05), but these were fewer in the OI group than the OC group. We found marked morphological changes in testicular tissues between the NC and OC groups, as well as between the OI and OC groups. We found that HFD induced obesity was detrimental to rat testes and that administration of IND ameliorated testicular changes caused by obesity.