Effect of silymarin and metformin on the sperm parameters and histopathological changes of testes in diabetic rats: An experimental study

Background: Oxidative stress is a major contributor to diabetes, which can lead to testicular damage and infertility. Objective: This study aimed to compare the effects of metformin as a chemical drug with silymarin as an herbal agent on the sperm parameters and histopathological changes of testes in diabetic rats. Materials and Methods: Thirty-two male Wistar rats (250-270 gr) were randomly divided into four groups: 1) control; 2) diabetic; 3) diabetic+metformin 200 mg/kg; and 4) diabetic+silymarin 100 mg/kg. Daily injections were administered intraperitoneally for 56 days. At the end of the treatment, blood sampling was performed for biochemical assessment. Then, the rats were sacrificed and their left testis and epididymis were cut for sperm analysis as well as histopathology and morphometric evaluation. Results: Diabetes was associated with a reduced sperm count, motility, viability, maturity, and chromatin quality of sperm (p ≤ 0.001). It was also associated with a higher malondialdehide level and lower total antioxidant capacity level of serum in comparison with the control group (p ≤ 0.001). There was a significant difference in the seminiferous tubule diameter, germinal epithelium height, and testicular histopathological alterations in the diabetic rats compared with the control rats (p ≤ 0.001). Treatment with metformin and silymarin improved the above-mentioned parameters and this improvement was more substantial in silymarin-treated animals (p ≤ 0.001). Conclusion: In diabetic rats, metformin and silymarin improved sperm parameters, sperm DNA integrity, seminiferous tubule diameter, germinal epithelium thickness, and testicular histopathological complications; this improvement was more substantial in the silymarin-treated group. So, the findings of this study suggest that silymarin is more effective than metformin in treating diabetic-induced infertility. Key words: Diabetes, DNA damage, Metformin, Silymarin, Sperm, Testis.

Morphological and morphometric adaptations of testes in broilers induced by glucocorticoid

Glucocorticoids (GCs) cause excess fat accumulation, which leads to fertility dysfunction in broilers. The study investigated alterations in the morphology and morphometry of the testes of broilers in response to GC and dexamethasone (DEX). Male day-old chicks were randomly divided into a control group and three experimental groups (E1, E2, and E3). The control group was fed a commercial broiler ration. The experimental groups were fed a commercial broiler ration containing GC (i.e. DEX 3, 5, and 7 mg/kg, respectively). The testes were collected and stained with haematoxylin and eosin to count the number of testicular seminiferous tubules. An increase in the seminiferous tubule count was initially seen, which declined as both the age of the broilers and the dose of DEX increased. Morphometric measurements, i.e., the testicular capsule thickness, seminiferous tubule diameter, and seminiferous epithelium height, were performed. The initial thickening of the testicular capsule was evident. There was a depletion of the interstitial (Leydig) cell population in the experimental groups with the age and increased with the dose advancement. The diameter of the seminiferous tubules and testicular capsule thickness remained upregulated in the treatment groups with the increased dose of DEX. The initial height of the seminiferous epithelium increased in the experimental groups of broilers. The study suggests that DEX greatly alters the morphological architecture of broiler testes; as a result, it could be said that DEX has the effect on the infertility of the broiler by affecting the morphology as well as the functionality of the testes.

Protective effects of curcumin against methotrexate-induced testicular damage in rats by suppression of the p38-MAPK and nuclear factor-kappa B pathways

Objective: Methotrexate (MTX), is a commonly used chemotherapeutic agent. This study aimed to investigate the possibility that curcumin (CMN) protects against MTX-induced testicular damage by affecting the phospho (p)-p38 mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-kB) signaling pathways. Methods: Male Wistar albino rats were subdivided into three groups. The control group was given an intragastric (ig) administration of dimethyl sulfoxide (DMSO) daily for 14 days, the MTX group was given a single intraperitoneal (ip) dose of MTX (20 mg/kg) on the 11th day, and the MTX+CMN group was given ig CMN (100 mg/kg/day, dissolved in DMSO) for 14 days and a single ip dose of MTX (20 mg/kg) on the 11th day. Results: The animal weights, the seminiferous tubule diameter, and germinal epithelium height significantly decreased in the MTX group compared to the control group. The testes weight and the ratio of the testes to body weight did not change, whereas the number of seminiferous tubules and the interstitial space width increased significantly in the MTX group. The number of phospho-p38 (p-p38) MAPK immunopositive cells and the immunoreactivity of NF-kB also increased in the MTX group compared to the control group. Conclusion: CMN prevented the MTX-induced decreases in the body weight, seminiferous tubule diameter, and the germinal epithelium height, while significantly reducing the number of histologically damaged seminiferous tubules and the interstitial space width changes due to MTX. CMN also reduced the number of p-p38 MAPK immunopositive cells and the NF-kB immunoreactivity.

The Effect Of Intrauterine Hypoxia On Testicular Reproductive Function

Goal — to assess the effect of antenatal hypoxia of various origins on the morphology and reproductive function of the testes of newborn and mature rats in experiment. Material and Methods — In experiments 15 white outbred female rats aged 4 to 10 months with a weight of 200±30 g were used. Laboratory animals were randomly divided into 2 experimental and 1 control groups, 5 females each. The first group underwent normobaric hypoxia throughout pregnancy (21 days). Hypoxia modeling was conducted in accordance with the method of N.N. Karkishchenko (2010). The second group underwent hemic hypoxia during the second and third week of pregnancy, in accordance with the method of L.M. Sosedova (2012). The third (control) group was not exposed to any effect throughout pregnancy. Results — in the testicles of newborn rats of the experimental groups, the decrease of tubule diameter was observed, the increase of stroma area and development of interstitial edema were noted. In the group of hemic hypoxia, a significant decrease in the number of Leydig cells was noted. In the tissues of the testicles of mature rats, who underwent antenatal hypoxia, a decrease of tubule diameter, a significant decrease in the spermatogenesis index and a decrease of spermatogonia number were noted. The developed damage of spermatogenic epithelium in experimental groups of newborns and mature rats was confirmed by marked expression of the apoptosis marker (Bax), weak expression of proliferation markers (Ki-67) and receptor of receptor of fibroblast growth factor (FGFR). Conclusion — in animals with chronic hypoxia of various origins, there is an inhibition of spermatogenesis and a violation of the spermatogenetic function of the testicular seminiferous tubules.

GONADAL AND EXTRAGONADAL SPERM RESERVES AND TESTICULAR HISTOMETRY OF POST PUBERTAL MUTURU BULLS.

Testes and epididymides were obtained from three pubertial Muturu bulls aged 11 months and 12 post pubertal bulls aged between 13 and 17 months and weighed between 91 and 148kg. Samples of each testis and epididymal segments namely, caput, corpus and cauda, were processed for histology while samples for gonadal and extragonadal sperm reserves were processed using the homogenisation technique. Paired testes weight increased from 78.99+8.74g at 11months of age to 90.10+10.86g. 110.30+5.67g and 106.34-6.52g at 13, 15 and 17 months of age, respectively. Paired testes sperm numbers were 3.00+1.50 X 10', 5.25 0.51 x 10', 10.44+2.55 x 10° and 9.14+3.08 x 10°, during the corresponding periods. Similarly, daily sperm production (DSP) increased by 77%, 250% and 207% at 13, 15 and 17 months of age, respectively. There was a 22% increase in the seminiferous tubule diameter from the time of puberty at 11 months to the time bulls were 15 months old, but the histological characteristics of the different epididymal segments did not follow any definite trend during these periods. Correlations between paired testes weight and other testicular and epididymal parameters were positive and significant (P<0.01). These results suggest that Muturu bulls may be used for breeding from the age of 15 months.

Testicular features of semi-confined Nellore bulls subjected to immunocastration

This study aimed to evaluate the effect of immunocastration on the scrotal circumference, as well as in the macroscopic and microscopic testicular features in Nellore bulls bred under a semi-confining system for 91 days. Sixty animals were divided into two groups: 30 intact animals (non-vaccinated) and 30 immunocastrated animals. The immunocastrated animals were treated with two anti-GnRH vaccine doses (BoprivaTM - Zoetis Australia Pty Ltd). The scrotal circumference (SC) was measured on days 0 and 56. Testicular parameters: biometry traits (length, width, and height – cm; volume – cm3 ) and weight (g) was measured on day 91. Three testicles from each group were histologically processed to find the seminiferous tubule diameter and epithelium thickness (µm). Data were analyzed in the Minitab® 19 statistical software. The macroscopic features (SC, and testes weight, volume, length, width, and height) and the microscopic features (seminiferous tubule diameter and epithelium thickness) were compared between the two groups through Student’s t-test and Mann-Whitney test. All the values were lower (p<0.05) in animals immunized against GnRH, except for the right testis width and seminiferous tubule diameter. Results indicate that immunization against GnRH affected testicular development.

The outcome of potassium dichromate exposure on histological structure of male rat sexual organs in suckling period

The aim of this study was the evaluation of integrity biomarkers of reproductive toxicity: histoarchitecture of genital organs (testis and epididymis) and sexual accessory glands (prostate, seminal vesicles and bulbo-urethral glands) at sexual maturity after exposure to potassium dichromate (Cr VI) in suckling period. Consequent to the exposure of male rat pups to hexavalent chromium during suckling period structural changes appeared in genital organs and sexual accessory glands, such as: basal membrane and epithelial disintegration, interstitial edema, wavy basal membrane due to a reduction in tubule diameter, Leydig cell necrosis; epithelial smoothing, epithelial necrosis; epithelial cells desquamation and falling in lumen. Exposure to potassium dichromate (Cr VI) during suckling period produced severe congestive and degenerative lesions in genital organs and sexual accessory glands.

Deletion of the PDZ-binding kinase (Pbk) gene does not affect male fertility in mice

The PDZ-binding kinase (PBK) protein is localised exclusively in spermatogenic cells, such as spermatogonia, spermatocytes and round spermatids, of the adult testis. However, its role in male fertility remains unknown. Analysis of adult Pbk-knockout (KO) male mice showed no significant difference in the weight of the testes, epididymis and seminal vesicle compared with adult wild-type (WT) mice. There were no significant differences in testis morphology, tubule diameter and the number of offspring born to females mated with KO or WT male mice. Sperm number, motility and morphology did not differ significantly between KO and WT mice. The oocyte fertilisation rate and embryo development following IVF were comparable between groups fertilised using spermatozoa from KO versus WT mice (P&gt;0.05). Further analysis revealed that the phosphorylation of the mitogen-activated protein kinases (MAPKs) p38 kinase, c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinases was dysregulated in the testis of KO mice. In conclusion, Pbk-KO male mice are fertile and their spermatozoa and testis do not show any morphological and functional abnormalities despite the dysregulated phosphorylation of MAPKs. It is likely that functional redundancy of PBK and overlapping substrate specificities of the MAPK superfamily compensated for the loss of PBK from the testis.

PSX-5 Reproductive organ weights, morphology and sperm reserves in intact and short scrotum post-pubertal male dairy sheep lambs

This study evaluated the use of short-scrotum castration to create sterile ram lambs. East Friesian x Lacaune male lambs born in February on a commercial sheep dairy were left intact (n = 19; IN) or rendered short-scrotum (n = 17; SS) within 10 d of birth. At 3 mo of age lambs were transferred to the study site and grazed as one group, while being supplemented with a grain mixture at 0.7 kg/d. Lambs were harvested at 6 mo of age, and reproductive tract of 5 IN and 5 SS lambs collected at random and shipped cold overnight for processing. Testis and epididymis were weighed. Segments of the left testis and epididymis were processed for histology, while a subsample of testicular parenchyma and epididymal segments of the right side were used to determine sperm reserves by tissue homogenization. Differences between testis types were determined using GLM of SAS. Testes and epididymal weights were reduced (P &lt; 0.001) in SS (115 and 22 g) compared to IN (364 and 50 g) lambs. Seminiferous tubule diameter was also smaller (P &lt; 0.001) in SS (142 µm) than IN (234 µm) lambs. Daily sperm production per gram testicular parenchyma (DSPG) was not different between types (3.8 vs. 2.3 million sperm/g; P = 0.06), but total daily sperm production was lower (P &lt; 0.01) in SS (0.23 billion) than IN (1.34 billion) ram lambs. Considerable variation was noted in DSPG in IN lambs suggesting still incomplete development at this age, and accounting for the lack of a significant difference. Epididymal sperm reserves were markedly reduced (P &lt; 0.001) in SS (0.1 billion) compared to IN (10.5 billion), and decreased along the epididymis, supported by evidence of phagocytic action by the epididymal epithelium. Data suggest that ram lambs rendered short-scrotum shortly after birth develop smaller testes with reduced sperm production, making them effectively infertile.

Effects of a standardized 40% ellagic acid pomegranate (Punica granatum L.) extract on seminiferous tubule histopathology, diameter, and epithelium thickness in albino Wistar rats after heat exposure

Background and Aim: It has long been known that the spermatogenic tissue is very sensitive to temperatures higher than its physiologic temperature and causing cessation of activity and resulting in sterility. This study investigated the effect of a standardized 40% ellagic acid extract of pomegranate on the histopathology, diameter, and epithelial thickness of seminiferous tubules in albino rats exposed to heat. Materials and Methods: Twenty-five male albino Wistar rats were randomized at 7-8 months of age to five treatment groups. Group C was not treated; Group T0 was treated with 0.5% of Na carboxymethyl cellulose (CMC) 2 ml/day and exposed to heat. T1, T2, and T3 were treated with 75, 150, and 300 mg/kg/day of a standardized 40% ellagic acid extract of pomegranate (Punica granatum L.), respectively. The animals were orally administered Na CMC or pomegranate extract and were exposed to sunlight for 15 min at 40°C-42°C for 14 days. The animals were sacrificed on day 15 and the testes were removed for histological evaluation and measurement of seminiferous tubule diameter and epithelium thickness. Results: The diameter of seminiferous tubules from rats exposed to heat and treated with 300 mg/kg/day pomegranate extract was larger and the epithelia thicker than those in the other groups (p= <0.05). The protective effects of the standardized 40% ellagic acid extract may have been mediated by its antioxidant activity. Conclusion: Compared with controls, administration of 300 mg/kg/day of a standardized 40% ellagic acid extract of P. granatum L. for 14 days increased seminiferous tubule diameter and epithelium thickness in albino Wistar rats exposed to heat. Keywords: diameter, epithelium thickness, heat, pomegranate extract, seminiferous tubule.