Increase In Testis Weight Research Articles

Bezafibrate alleviates diabetes-induced spermatogenesis dysfunction by inhibiting inflammation and oxidative stress

The metabolic disorders caused by diabetes can lead to various complications, including male spermatogenesis dysfunction. Exploring effective therapeutics that attenuate diabetes mellitus (DM)-induced male subfertility is of great importance. Pharmaceuticals targeting PPARα activation such as bezafibrate have been regarded as an important strategy for patients with diabetes. In this study, we use streptozocin (STZ) injection to establish a type 1 DM mice model and use bezafibrate to treat DM mice and evaluate the effects of bezafibrate on the spermatogenic function of the DM male mice. Bezafibrate treatment exhibited protective effects on DM-induced spermatogenesis deficiency, as reflected by increased testis weight, improved histological morphology of testis, elevated sperm parameters, increased serum testosterone concentration as well as increased mRNA levels of steroidogenesis enzymes. Meanwhile, testicular cell apoptosis, inflammation accumulation and oxidative stress status were also shown to be alleviated by bezafibrate compared with the DM group. In vivo and in vitro studies, PPARα specific inhibitor and PPARα knockout mice were further used to investigate the role of PPARα in the protective effects of bezafibrate on DM-induced spermatogenesis dysfunction. Our results indicated that the protection of bezafibrate on DM-induced spermatogenesis deficiency was abrogated by PPARα inhibition or deletion. Our study suggested that bezafibrate administration could ameliorate DM-induced spermatogenesis dysfunction and may represent a novel practical strategy for male infertility.

(068) Perfluorooctane Sulfonate (pfos) And Perfluorooctanic Acid (pfoa) Sub-chronic Exposure has Minimal Impact on Erectile Function in Sprague-dawley Ratsa

Abstract Introduction Per and polyfluoroalkyl substances (PFAS) are commonly used in industrial and consumer products and are environmental and health contaminants. The most common PFAS chemicals, perfluorooctane sulfonate (PFOS) and perfluorooctanic acid (PFOA), are used in firefighting foam, nonstick bakeware, water-resistant fabrics, grease-resistant paper and fire-resistant materials. PFAS is a known endocrine disruptor and can reduce testosterone and lower semen quality. The impact of PFAS exposure on erectile physiology is unknown. Objective Our study will assess the effect of PFOS and PFOA on erectile function in Sprague-Dawley rats. We hypothesize that PFOS and PFOA will decrease erectile function. Methods Adult male Sprague-Dawley rats were divided into three groups (n=10/group): 1) Control; 2) PFOS (10mg/kg/day); and 3) PFOA (10mg/kg/day). PFOS/PFOA was dissolved in 0.05% Tween 20 and administered in drinking water for 28 days. Controls received 0.05% Tween 20 in drinking water. The study was carried out in two different cohorts of rats (n=4 and n=6/group). Following 28 days of PFOS/PFOA exposure, rats were placed on regular drinking water for 2 weeks. Body weights were collected weekly and terminal organ weights of liver, testes, spleen, heart, lungs, brain, seminal vesicles, kidneys, bladder, and thymus were recorded. Erectile function was assessed prior to PFAS exposure and every 2 weeks thereafter via apomorphine-induced behavioral erections and yawns. Nerve-stimulated intracavernosal pressure and mean arterial pressure (ICP/MAP) was measured 2 weeks after exposure stopped. Results Four weeks of PFOS exposure significantly decreased body weight while PFOA exposure did not impact body weight (Con: 588±63.5g; PFOS: 453±16.3g; PFOA: 535±29.9g, p<0.01). The PFOS dose in the second cohort was overtly toxic with 5/6 rats dying following 3-4 weeks of exposure. Two weeks following PFOS/PFOA exposure, livers were markedly enlarged (Con: 19.4±2.85g; PFOS: 25.4±0.16g; PFOA: 20.2±3.77g, p<0.01), spleens were smaller (Con: 0.93±0.12g; PFOS: 0.65±0.03g; PFOA: 0.88±0.12g, p<0.05), and testes were larger in PFOS rats (Con: 1.8±0.07g; PFOS: 2.0±0.06g; PFOA: 1.8±0.07). No other organ weights were different across exposure groups. PFOS/PFOA exposed animals had similar numbers of apomorphine-induced erections throughout the 6 week period. Two weeks post-exposure, ICP/MAP was unchanged between all groups (n=5-10/group). Ongoing studies will assess testosterone levels. Conclusions A sub-chronic PFOS exposure markedly decreased body weight, increased liver size, decreased spleen weight, and increased testes weight. PFOA exposure did not impact body or organ weights. Following PFOS or PFOA exposure, there was no change in apomorphine-induced behavioral erections or ICP/MAP. Additionally, the dose of PFOS assessed was quite toxic to the second cohort of animals. Future studies will assess if a more chronic exposure will impact erectile function. Disclosure No.

Effects of Perilla frutescens Var. Acuta in Busulfan-Induced Spermatogenesis Dysfunction Mouse Model.

The leaves of Perilla frutescens var. acuta (PFA) are generally reported to have antioxidant, anti-allergic, anti-inflammatory, and antitumor effects and commonly used as a traditional medicine in East Asia. This study aimed to investigate the protective effect and antioxidant activity of PFA on busulfan-induced testicular dysfunction, histological damage, oxidative stress (OS), sperm quality, and hormone levels using a mouse model. C57BL/6 male mice were divided into four groups: control, busulfan-only treated, and varying concentrations of PFA (100 and 200 mg/kg) with busulfan. In the busulfan group, 40 mg/kg of busulfan was intraperitoneally injected to induce azoospermia. Mice were orally administered PFA for 35 consecutive days after busulfan administration. Samples were collected and assessed for testis/body weight, testicular histopathology, sperm quality, serum hormone levels, and OS to evaluate the effects of PFA treatment on spermatogenesis dysfunction induced by busulfan. The busulfan-induced testicular dysfunction model showed reduced testis weight, adverse histological changes, significantly decreased sex hormones and sperm quality, and attenuated OS. These results indicate that PFA treatment significantly increased testis weight, testis/body weight, epididymal sperm count, motility, and testosterone level compared with busulfan alone. PFA treatment also attenuated the busulfan-induced histological changes. Furthermore, compared with mice treated with busulfan alone, PFA supplementation upregulated the testicular mRNA expression of the antioxidant enzymes superoxide dismutase 1 (Sod1) and glutathione peroxidase 1 (Gpx1), with a decrease in malondialdehyde (MDA) production and an increase in SOD and GPx activities. This study shows that PFA exerts a protective effect against testicular damage by attenuating OS induced by busulfan. Our results suggest that PFA is a potentially relevant drug used to decrease the side effects induced by busulfan on testicular function and sperm during cancer chemotherapy.

Ameliorative Role of Allicin in Arsenic-Induced Liver and Gonad Apoptosis in Male <i>Swiss</i> Albino Mice (<i>Mus musculus</i>)

Arsenic toxicity is a matter of concern in the present-day world. Arsenic, a potentially hazardous metalloid easily gets biomagnified through the food chain and also affects not only the vital organs such as the kidney, and liver of human beings but also the reproductive organs of males. This study was designed to propose allicin, the main bio-active component of garlic to address arsenic toxicity more efficiently and without any side effects apart from the costly conventional chelation therapy which is not free from various side effects. To conduct this study, allicin has been quantified and collected from ethanolic extract of garlic by High-Performance Liquid Chromatography (HPLC) using C18 column at 254nm wave-lengths against standard allicin at the retention time of 18.775min. The calculated concentration of allicin is 77.80%. Next, arsenic trioxide (As2O3) (20mg/kg) and allicin (100mg/kg) were administered orally for toxicity and treatment respectively in mice (Mus musculus) for 30 days. Compared to the control group, liver marker enzymes i.e., Serum Glutamic Pyruvate Transaminase (SGPT), Serum Glutamic Oxaloacetic Transaminase (SGOT), and Alkaline Phosphatase (ALP) levels were significantly increased in the arsenic-induced group. On the other hand, co-treatment with allicin significantly recovered liver enzyme parameters to normal levels (p<0.05). Decreased weight of testis, sperm count, and increased numbers of Sperm Head Anomalies (SHA) indicate reduced reproductive potential in the arsenic-induced group of male albino mice. On the contrary, co-treatment with allicin significantly increased testis weight, sperm count and decreased SHA count (p<0.05). On examining histological slides of the liver and testis, normal histo-architecture was observed in both control and arsenic-induced allicin co-treated groups; whereas damage was observed in the arsenic-induced group. Generated data pointed out that allicin offers significant protection to the mammalian liver and male gonad (testis) against arsenicinduced toxicity.

Positive Effects of the Nutraceutical Association of Lycopene and Selenium in Experimental Varicocele.

Many natural substances commonly found in healthy diets have been studied for their potential to reduce male infertility associated with varicocele. A positive role of selenium (Se) or lycopene alone was demonstrated in experimental varicocele, while no data are available on their association. One group of male Sprague-Dawley rats was sham operated and daily treated with Se (3 mg/kg, i.p.), lycopene (1 mg/kg, i.p.), or their association. A second group underwent surgery to induce varicocele. Sham and half of the varicocele animals were sacrificed after twenty-eight days, while the residual animals were treated for one more month and then sacrificed. In varicocele animals, testosterone levels and testes weight were reduced, Hypoxia Inducible Factor-1α (HIF-1α) expression was absent in the tubules and increased in Leydig cells, caspare-3 was increased, seminiferous epithelium showed evident structural changes, and many apoptotic germ cells were demonstrated with TUNEL assay. The treatment with lycopene or Se alone significantly increased testis weight and testosterone levels, reduced apoptosis and caspase-3 expression, improved the tubular organization, decreased HIF-1α positivity of Leydig cells, and restored its tubular positivity. Lycopene or Se association showed a better influence on all biochemical and morphological parameters. Therefore, the nutraceutical association of lycopene plus Se might be considered a possible therapeutic tool, together with surgery, in the treatment of male infertility. However, long-term experimental and clinical studies are necessary to evaluate sperm quantity and quality.

Effect of corn straw or corncobs in total mixed ration during peri-puberty on testis development in Hu lambs

Corn straw and corncobs contain large amounts of crude fibers and are widely used in mutton sheep husbandry in northwest China. The aim of this study was to determine whether feeding with corn straw or corncobs affects lamb testis development. A total of 50 healthy Hu lamb at two-month-old (average body weight of 22.3 ± 0.1 kg) were randomly and equally divided into two groups, and the lambs were equally allocated to five pens in each group. The corn straw group (CS) received a diet containing 20% corn straw, whereas the corncobs group (CC) received a diet containing 20% corncobs. After a 77-day feeding trial, the lambs, except the heaviest and lightest in each pen, were humanely slaughtered and investigated. Results revealed no differences in body weight (40.38 ± 0.45 kg vs. 39.08 ± 0.52 kg) between the CS and CC groups. Feeding diet containing corn straw significantly (P < 0.05) increased testis weight (243.24 ± 18.78 g vs. 167.00 ± 15.20 g), testis index (0.60 ± 0.05 vs. 0.43 ± 0.04), testis volume (247.08 ± 19.99 mL vs. 162.31 ± 14.15 mL), diameter of seminiferous tubule (213.90 ± 4.91 μm vs. 173.11 ± 5.93 μm), and the number of sperm in the epididymis (49.91 ± 13.53 × 108/g vs. 19.34 ± 6.79 × 108/g) compared with those in the CC group. The RNA sequencing results showed 286 differentially expressed genes, and 116 upregulated and 170 downregulated genes were found in the CS group compared with the CC group. The genes affecting immune functions and fertility were screened out. Corn straw decreased the mtDNA relative copy number in the testis (P < 0.05). These results suggest that compared with corncobs, feeding corn straw in the early reproductive development stage of lambs increased the testis weight, diameter of seminiferous tubule and the number of cauda sperm.

Effects of Leptin antagonist treatments on testosterone and testis histological characteristics of immature male mice

The present study aimed to ascertain how leptin antagonist injection affected testis weights, testis morphology and testosterone levels in immature male Swiss mice. Animals were administered with anti-leptin antibody subcutaneously, with or without equine chorionic gonadotropin (eCG). Control animals were treated with non-immune serum. Blood and testis were collected. The Androgen profile was analyzed in serum and tissue homogenates, and testes were histologically examined. Compared to controls, mice treated with an anti-leptin antibody with or without gonadotropins had a significant (p&lt;0.05) increase in testis weight. Testosterone concentrations in the testis were significantly (p&lt;0.05) higher in mice administered with anti-leptin antibody compared to control, but testosterone concentrations in blood were not affected. The diameter of seminiferous tubules, the diameter of the lumen and the width of spermatogenic cells were significantly (p&lt;0.05) higher in mice in treatment groups compared to controls. We conclude that anti-leptin antibody administration in immature male mice increased testosterone concentrations in the testis and improved testis histological characteristics. Keywords: leptin; mouse; histology; testis; testosterone; immature male

Involvement of Hypoxia-Inducible Factor 1-α in Experimental Testicular Ischemia and Reperfusion: Effects of Polydeoxyribonucleotide and Selenium

Polydeoxyribonucleotide (PDRN) is an agonist of the A2A adenosine receptor derived from salmon trout sperm. Selenium (Se) is a trace element normally present in the diet. We aimed to investigate the long-term role of PDRN and Se, alone or in association, after ischemia-reperfusion (I/R) in rats. The animals underwent 1 h testicular ischemia followed by 30 days of reperfusion or a sham I/R and were treated with PDRN or Se alone or in association for 30 days. I/R significantly increased hypoxia-inducible factor 1-α (HIF-1α) in Leydig cells, malondialdehyde (MDA), phosphorylated extracellular signal-regulated kinases 1/2 (pErk 1/2), and apoptosis decreased testis weight, glutathione (GSH), testosterone, nuclear factor erythroid 2-related factor 2 (Nrf2), induced testicular structural changes, and eliminated HIF-1α spermatozoa positivity. The treatment with either PDRN or Se significantly decreased MDA, apoptosis, and HIF-1α positivity of Leydig cells, increased testis weight, GSH, testosterone, and Nrf2, and improved the structural organization of the testes. PDRN and Se association showed a higher protective effect on all biochemical, structural, and immunohistochemical parameters. Our data suggest that HIF-1α could play important roles in late testis I/R and that this transcriptional factor could be modulated by PDRN and Se association, which, together with surgery, could be considered a tool to improve varicocele-induced damages.

Methanol Extract of Azanza garckeana Fruit Pulps Protects against Formalin-Induced Reproductive Toxicity in Adult Albino Male Mice

Introduction: The aqueous extract of Azanza garckeana was recently reported of exhibiting ameliorative and pro-fertility properties however the protective effects on formalin testicular toxicity have not been studied.&#x0D; Objective: This study investigated the protective effect of methanol extract of Azanza garckeana on formalin-induced testicular toxicity.&#x0D; Methods: Forty male albino mice were randomly divided into 8 groups of 5. Animals in the first group (1) served as control and administered normal saline (1 ml/kg) by the oral route daily for 40 days. In similar manner, animal in groups 2 received formalin (10 mg/kg) by the IP route, while animals in groups 3; 4 and 5 concurrently received formalin (10 mg/kg IP) and extract at doses of 125; 250 and 500 mg/kg respectively by the oral route. Mice in groups 6; 7 and 8 received the extract at doses of 125; 250 and 500 mg/kg respectively. Phytochemical analysis was conducted for each constituent using specific methods. Gonadotropin and sperm analysis were carried out using standard methods.&#x0D; Result: Phytochemical screening revealed the presence of various constituents, but notably flavonoids. Induced-toxicity with formalin and concurrent treatment with extract at doses of 250 and 500 mg/kg from day 20 to 40 caused significant body weight increase compared to baseline (p &lt; 0.05). Similarly, treatment with the extract alone at all doses caused significant increase in body weight from day 20 to 40 (p &lt; 0.05). Treatment with the extract at 250 and 500 mg/kg, caused a significant increase in weight of testes and epididymis compared to control and untreated group (p &lt; 0.05).The extract induced significant increase in gonadotropin levels of animals compared to control and the untreated group (p &lt; 0.05).The extract at 125 mg/kg demonstrated the highest fecundity potential, but there was no any consistent relationship between GSI and fecundity.&#x0D; Conclusion: This investigation was able to establish the protective and pro-fertility potentials of methanol extract of Azanza garckeana.

Icariin improves testicular dysfunction via enhancing proliferation and inhibiting mitochondria-dependent apoptosis pathway in high-fat diet and streptozotocin-induced diabetic rats

BackgroundDiabetes mellitus (DM), a chronic metabolic disease, severely impairs male reproductive function. However, the underpinning mechanisms are still incompletely defined, and there are no effective strategies or medicines for these reproductive lesions. Icariin (ICA), the main active component extracted from Herba epimedii, is a flavonoid traditionally used to treat testicular dysfunction. Whether ICA can improve male reproductive dysfunction caused by DM and its underlying mechanisms are still unclear. In this study, by employing metformin as a comparative group, we evaluated the protective effects of ICA on male reproductive damages caused by DM and explored the possible mechanisms.MethodsRats were fed with a high fat diet (HFD) and then intraperitoneally injected with streptozotocin (STZ) to induce diabetes. Diabetic rats were randomly divided into T2DM + saline group, T2DM + metformin group and T2DM + ICA group. Rats without the treatment of HFD and STZ were used as control group. The morphology of testicular tissues was examined by histological staining. The mRNA expression levels were determined by quantitative real-time PCR. Immunostaining detected the protein levels of proliferating cell nuclear antigen (PCNA), hypoxia-inducible factor 1-alpha (HIF-1α) and sirtuin 1 (SIRT1) in testicular tissues. TUNEL assay was performed to determine cell apoptosis in the testicular tissues. The protein expression levels of HIF-1α and SIRT1 in the testicular tissues were determined by western blot assay.ResultsICA effectively improved male reproductive dysfunction of diabetic rats. ICA administration significantly decreased fasting blood glucose (FBG) and insulin resistance index (IRI). In addition, ICA increased testis weight, epididymis weight, sperm number, sperm motility and the cross-sectional area of seminiferous tubule. ICA recovered the number of spermatogonia, primary spermatocytes and Sertoli cells. Furthermore, ICA upregulated the expression of PCNA, activated SRIT1-HIF-1α signaling pathway, and inhibited intrinsic mitochondria dependent apoptosis pathway by upregulating the expression of Bcl-2 and downregulating the expression of Bax and caspase 3.ConclusionThese results suggest that ICA could attenuate male reproductive dysfunction of diabetic rats possibly via increasing cell proliferation and decreasing cell apoptosis of testis. ICA potentially represents a novel therapeutic strategy against DM-induced testicular damages.

Dietary supplementation of grape seed tannin extract stimulated testis development, changed fatty acid profiles and increased testis antioxidant capacity in pre-puberty hu lambs

Grape seed tannin extract (GPE) from wine grape pomace has many effective anti-oxidative effects and is used as a promising natural feed additive in the animal feed industry. This study investigated the effect of GPE as a source of tannin on the antioxidant capacity and testis development in Hu lambs. Twenty-seven 3-month-old ram lambs were randomly assigned to three groups. For each treatment group, nine lambs were allocated to nine pens (one lamb per pen). The lambs in the control group were fed a control diet without GPE for 61 days from D21 to D80. Group I (TAN1) was fed with 0.36% GPE diet, and Group II (TAN2) was fed with 0.72% GPE diet. After an 81-day feeding trial, all lambs except the heaviest and lightest in each group were humanely slaughtered and investigated. Results showed that feeding GPE did not affect the body weight, average daily gain, dry matter intake, scrotal circumference, and testis index. Meanwhile, feeding with 0.36% GPE diet increased testis weight, testis volume, and epididymis weight (P ≤ 0.05) compared with those of the control, but no difference was found between TAN1 and TAN2 groups. Copper-zinc superoxide dismutase (Cu-ZnSOD), steroid acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), follicle-stimulating hormone receptor (FSHR), elongation of very long chain fatty acid protein 2 (ELOVL2), fatty acid desaturase (FADS2), and proliferating cell nuclear antigen (PCNA) mRNA in TAN1 and TAN2 groups were significantly up-regulated (P < 0.05). GPE also markedly increased the antioxidant status of testis. Compared with the control group, the treatment groups showed significantly increased superoxide dismutase (SOD) activity (314.23 ± 18.64 U/mg prot in control, 505.22 ± 63.47 U/mg prot in TAN1 and 587.88 ± 55.94 U/mg prot in TAN2, P < 0.05) and total antioxidant capacity (T-AOC) (98.23 ± 18.99 U/g prot in control, 202.15 ± 34.19 U/g prot in TAN1 and 189.57 ± 18.95 U/g prot in TAN2, P < 0.05). Consuming 0.72% GPE also changed the fatty acid profiles in testis with increased C15:1, C22:6n3, and total n-3 fatty acids (P < 0.05) but decreased C22:5n3 (P < 0.05). Therefore, feeding lambs with GPE stimulated testis seminiferous tubule development and increased the number of Sertoli cells (10.56 ± 0.44 in control, 14.10 ± 0.57 in TAN1 and 13.60 ± 0.42, P < 0.05), and seminiferous tubule diameter (109.30 ± 4.56 μm in control, 164.49 ± 5.37 μm in TAN1 and 146.56 ± 4.53 μm in TAN2, P < 0.05). These results suggested that feeding GPE in the early reproductive development stage of lambs upregulated the expression of antioxidative, steroidogenesis, and polyunsaturated fatty acid metabolism-related genes, changed the fatty acid profiles, increased the antioxidant capacity in lamb's testis, and contributed to testis development and spermatogenesis.

Seasonal change in male reproductive investment of a fish

Many animals are sexually dimorphic, whereby males may display brighter body coloration and more distinctive ornamentation than females. Fishes in temperate regions markedly change their energy allocation toward reproduction in response to the seasonal environment. Seasonal change in reproductive investment affects the expression of sexually dimorphic traits in males through gonadal weight change. Here, we report seasonal changes in body size, testis weight and sexual dimorphism of the fins (anal fin length and dorsal fin length) in the northern medaka Oryzias sakaizumii. Fish were collected periodically from a wild population (Aomori). Gonad weight increased from May to July in both males and females, corresponding to the reproductive season. Moreover, during this period, the degree of sexual dimorphism in fin length increased. To investigate the relationship between testis weight and individual differences in male fin length, we analyzed relationships among morphological traits using structural equation modeling. In the reproductive season, increased testis weight was associated with longer fin length in males, but the relationship disappeared after the reproductive season. These observations suggest that the sexually dimorphic fin in this fish is a mating signal. Results from a mating experiment also support this view. Males with larger size and/or longer fin attracted more mates than those with smaller fins, suggesting that sexual selection operates through females choosing to mate with males having longer fins.

Aromatase Inhibitor, Flax seed and Sage Effects on Adult Rats Fertility and Sexual Behavior

The current study was designed to detect estrogen role in adult male rats by giving aromatase inhibitor ( ltrozole 1 mg̷ Kg. B. W) for 60 days orally to inhibit aromatase enzyme activity and to study the effect of giving source of estrogen flax seeds (25 g ̷ 100 g diet) and aqueous extract sage (1000mg ̷ Kg. B.W.) orally. The study showed that adult male rats treated with ltrozole resulted in a significant decrease in the weight of testes, prostate , percentage of live sperms , number of sperms , number of mounting , intromission, ejaculation and in the time period from mixing with females to the first mounting , intromission and ejaculation , the time period between mount and another, intromission and another, ejaculation and another, and significant increase in the percentages of dead as well as abnormal sperms. Flax seeds treatment significantly reduced percentage of live sperms, number of sperms, number of mounting , intromission, and ejaculation, and significantly increased percentage of dead sperms compared with control group. Sage extract treatment caused a significant decrease in number of sperms, number of mounting, intromission and ejaculation , as well as a significant increase in the time period from mixing of female to the first mounting, first intromission and first ejaculation compared with control. The results showed that administration of sage extract with letrozole showed a significant increase in testis weight, percentages of live sperms and number of sperms, and asignificant decrease in dead and abnormal sperms compared with the letrozole group. Administration of flax seed with letrozole did not significantly change weight of the testis , prostate, percentage of live, dead , abnormal sperm and number of sperm compared with letrozole group. Administration of Flax seed and sage extracts to male rats treated with letrozole did not significantly change all the done tests of sex behavior. The study concluded that estrogen has an important role in the function of male reproductive system, which is shown through the negative effects of the aromatase inhibitor on the weights of the testis, prostate, live and dead and abnormal sperm, and sexual behavior. Sage extract, but not flax seed, has benefit effects on the defect induced by aromatase inhibitor letrozole both flax seed and sage do not produce positive effects on sexual behavior .

Prepubertal PTU treatment in rat increases Sertoli cell number and sperm production.

The number of Sertoli cells (SCs) ultimately determines the upper limit of sperm production in the testis. Previous studies have shown that thyroid hormones (TH) receptors are abundantly expressed in developing SCs; therefore, it was highly significant to discover that transient neonatal hypothyroidism induced by the goitrogen 6-n-propyl-2-thiouracil (PTU) can extend SCs proliferation beyond the first 2 weeks postnatal and increase testis weight and sperm production. Further studies concluded that treatment must begin before day 8 post birth in rats. Recent studies, however, showed that SCs present in the transition region at the rete testis exhibit a more immature phenotype and have prolonged mitotic activity, which led to the hypothesis that SCs in this region will retain the capacity to respond to PTU treatment over a longer period of time. In the present study, male Wistar rats were treated with PTU from days 21 to 40 and were evaluated at 40 and 160 days of age. Similar to neonatal rat SCs, it was demonstrated that prepubertal SCs in the transition region have a high mitotic activity and are highly sensitive to TH levels. This delayed, transient hypothyroidism resulted in significantly increased testis weight, SCs number and daily sperm production. The results demonstrate for the first time that Sertoli cells showing plasticity in the transition region can be stimulated to increase proliferation and contribute to a late stage surge in testis weight and sperm output.

Effects of quantitative trait loci determining testicular weight in DDD/Sgn inbred mice are strongly influenced by circulating testosterone levels.

ObjectiveTesticular growth and development are strongly influenced by androgen. Although both testis weight and plasma testosterone level are inherited traits, the interrelationship between them is not fully established. Males of DDD/Sgn (DDD) mice are known to have extremely heavy testes and very high plasma testosterone level among inbred mouse strains. We dissected the genetic basis of testis weight and analyzed the potential influence of plasma testosterone level in DDD mice.MethodsQuantitative trait loci (QTL) mapping of testis weight was performed with or without considering the influence of plasma testosterone level in reciprocal F2 intercross populations between DDD and C57BL/6J (B6) mice, thereby assessing the influence of testosterone on the effect of testis weight QTL. Candidate genes for testis weight QTL were investigated by next-generation sequencing analysis.ResultsFour significant QTL were identified on chromosomes 1, 8, 14, and 17. The DDD-derived allele was associated with increased testis weight. The F2 mice were then divided into two groups according to the plasma testosterone level (F2 mice with relatively “low” and “high” testosterone levels), and QTL scans were again performed. Although QTL on chromosome 1 was shared in both F2 mice, QTL on chromosomes 8 and 17 were identified specifically in F2 mice with relatively high testosterone levels. By whole-exome sequencing analysis, we identified one DDD-specific missense mutation Pro29Ser in alpha tubulin acetyltransferase 1 (Atat1).ConclusionMost of the testis weight QTL expressed stronger phenotypic effect when they were placed on circumstance with high testosterone level. High testosterone influenced the QTL by enhancing the effect of DDD-derived allele and diminishing the effects of B6-derived allele. Since Pro29Ser was not identified in other inbred mouse strains, and since Pro29 in Atat1 has been strongly conserved among mammalian species, Atat1 is a plausible candidate for testis weight QTL on chromosome 17.

Pemberian Fraksi Daun Kemangi (Ocimum americanum L.) terhadap Spermatozoa Tikus Putih Jantan (Rattus norvegicus)

The major cause of a spermatogenetic process disorder in testis is free radical. One of the traditional medicines commonly used as an aphrodisiac is Kemangi plant. Rosary Basil leaf ( Ocimum americanum L.), Indonesian name is Kemangi, contains active substances, such as flavonoid, saponin, tannin, and triterpenoid/steroid which is antioxidant and fertility. The purpose of the present research was to analyze the effect of testis weight, diameter, epithelium thickness, morphology, and spermatozoa amount of male white mouse ( Rattus norvegicus ) after feeding fractions of Kemangi leaf ( Ocimum americanum L.) This study was experimental with a posttest-only control group design. The research sample size was 24 furrow male white mice Sprague Dawley, 60-74 days old. They were divided into 4 treatment groups, each being fed the fractions of n-hexane, ethyl acetate, and water-ethanol of Kemangi leaf at a dosage of 100 mg/mouse/day, compared to CMC 1% for the control group for 36 days, and then a dissection was done to take out secreted substances and to examine the histology of testis. The result showed that there were increases of testis weight in the treatment group as compared to that of the control (p=0.018). In addition, there were increases in diameter and thickness of seminiferous tubule epithelium of testis, in the morphology of abnormal sperm (p=0.000), and in spermatozoa amount (p=0.020). Fractions of Kemangi leaf can increase testis weight, diameter, and epithelium thickness, morphology, and spermatozoa amount of furrow male white mice Sprague Dawley, particularly the fraction of ethyl acetate.

Leaf-Methanolic Extract of Pseudopanax arboreus (Araliaceae) (L. F. Phillipson) Reverses Amitriptyline-Induced Sexual Dysfunction in Male Rats

The people of the Bayangi tribe (Manyu Division) of Cameroon have used the leaves of Pseudopanax arboreus (Araliaceae) traditionally for decades as an aphrodisiac. In order to scientifically investigate this folk claim, we evaluated the effects of the leaf-aqueous extract of P. arboreus on the sexual behavior of normal male rats. The present study was designed to assess the effects of the leaf-methanolic extract of P. arboreus on amitriptyline-induced sexual dysfunction in male rats. Sexually impaired male rats were randomly divided into 4 groups of 8 rats each. Group 1 received 10 ml/kg distilled water, while group 2 was given 6 mg/kg Viagra. Groups 3 and 4 received 46.5 and 93 mg/kg of the leaf-methanolic extract, respectively. Female rats were made receptive by ovariectomy and subsequent hormonal treatment. Sexual behavior parameters were monitored on days 1, 7, 14, and 21 by pairing each male to a receptive female. The extract-treated rats registered significant decrease in mount latency (ML) and intromission latency (IL); nonsignificant increase in mount frequency (MF), intromission frequency (IF), and penile licking (PL); significant decrease in postejaculatory interval (PEI); contrasting effects in ejaculation latency (EL); and mean intromission interval (MII). Both doses of the extract also provoked a significant increase in relative weight of testes, but had no significant effect on the plasma hormonal profile. These findings are similar to those obtained with normal male rats and show that the leaf-methanolic extract of P. arboreus could constitute a potential solution to male sexual impairment.

Neonatal exposure to T3 disrupts male reproductive functions by altering redox homeostasis in immature testis of rats.

T3 is the active hormone, produced by peripheral deiodination of thyroxine. Exposure to excess thyroid hormones leads to hypermetabolic state and thus generates oxidative stress which seems to be involved in hyperthyroidism-induced testicular pathophysiology. We investigated the effects of T3 administration on the testis during development throughout sexual maturation in rats. Male pups were divided into two groups. T3 group was administered 80µg/kg body weight intraperitoneal T3 injections daily for 21days from the 1st postnatal day, while the control group was administered saline intraperitoneal injections. The pups were sacrificed at pnd 10, 20 and 30. T3 treatment resulted in a significant decrease in body weight at all ages tested and an increase in testis weight during the treatment period. The treatment produced imbalance in their testicular redox status, reflected by a significant increase in the amount of thiobarbituric acid-reactive substances and protein carbonyl content in the testicular homogenates of 20-day-old rats. We observed a significant increase in antioxidant system activities γ-glutamyl transferase, glucose-6-phosphate dehydrogenase, catalase and superoxide dismutase, reduced glutathione content and lactate dehydrogenase activity. Histological examination showed altered seminiferous tubules, degenerated germ cells and decreased height of the germinal epithelium. Chronic neonatal exposure to T3 resulted in redox state alterations which contribute to testicular impairment.

Effects of astaxanthin on biochemical and histopathological parameters related to oxidative stress on testes of rats on high fructose regime.

Astaxanthin (ASX) is a xanthophyll family of hydroxycarotenoids which contains several double bonds. It is produced by Haemococcus pluvialis, a microalgae and possesses antioxidant and anti-inflammatory properties. The aim of this study was to test whether ASX could protect against oxidative damage in the testicular tissues of rats receiving high fructose. The rats (n=24) were randomly divided into two main groups: control and fructose (30%, via drinking water) and then each main group either not supplemented or supplemented with ASX (1mg kg-1 day-1 , within 0.2ml olive oil) via oral gavage. Data were subjected to two-way ANOVA. High fructose consumption tended to increase testis weight and serum testosterone concentration and decreased testicular tissue glutathione-S-transferase (GST) and superoxide dismutase (SOD) levels, but did not affect testicular tissue malondialdehyde (MDA) concentration. Astaxanthin administration increased testosterone, GST and SOD levels and testis weight and decreased MDA concentration. However, ASX administration did not reverse alterations in antioxidant parameters caused by high fructose consumption. Inducible nitric oxide synthase (iNOS) tended to increase in sertoli cell, spermatid and spermatogonia, but not in spermatocytes and leydig cell in response to high fructose consumption. Astaxanthin administration tended to reverse elevation in iNOS in testis cells. In conclusion, ASX could help alleviate oxidative damage caused by high fructose consumption.

Asiatic acid attenuates high-fat diet-induced impaired spermatogenesis.

Testicular cell apoptosis is associated with impaired spermatogenesis. It has been reported that Asiatic acid (AA) may suppress apoptosis. However, little is known about the effect of AA on high-fat diet (HFD)-induced impairment of spermatogenesis. The aim of the present study was to determine whether AA protects against HFD-induced impairment of spermatogenesis. Sprague-Dawley rats were randomly divided into three groups: Control group, HFD group and AA (50 mg/kg) + HFD group. Rats fed an HFD were orally administered with AA (50 mg/kg) daily for 12 weeks, and blood samples, testis and epididymis were harvested for further analysis. Sex hormones were detected and hematoxylin and eosin staining was performed to examine the morphological changes of the testis. Semen samples were collected to evaluate sperm quality and apoptosis was determined. The results indicate that AA treatment significantly increased testis weight, testis/body weight, spermatogonia, Leydig cells and Sertoli cells in the testis of obese mice (P<0.05). AA treatment also attenuated HFD-induced histological change. AA treatment prevented HFD-induced decrease of sex hormones and the quality of semen samples (P<0.05). Furthermore, HFD-induced apoptosis was significantly attenuated by AA treatment (P<0.05). In conclusion, the results suggest that AA is able to ameliorate HFD-induced impaired spermatogenesis via inhibiting apoptosis in Sprague-Dawley rats. AA may have therapeutic value in the treatment of obesity-related impairment of spermatogenesis.