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.

Effect of salicylidene salicylhydrazide on testes of albino mice: A histomorphological study.

Objectives: To evaluate salicylidene salicylhydrazide (SCS) for any possible toxicological effects on male reproductive system in Albino male BALB/c mice. Study Design: Experimental study. Study Setting: Department of Histopathology, Khyber Medical College Peshawar and Department of Pharmacy, University of Peshawar, Peshawar were utilized for conducting this study. Period: July, 2020 to December, 2020. Material & Methods: Male BALB/c mice were daily administered with SCS at 5, 25, and 50 mg/kg for 7 and 14 days. The body and testes weights were measured and the testes were subjected to histological tissue processing techniques. Results: The extent of testicular toxicity was evaluated by using modified Johnsen scoring system for assessing the level of spermatogenesis, and morphometric analysis by measuring the diameter of the seminiferous tubules, thickness of germinal epithelium, and area of interstitial cells of Leydig. The body and testes weights showed that the various doses of SCS have no substantial effects after 7 and 14 days. The tested doses of SCS did not produce any distinguishable change in the normal histological features of seminiferous tubules and interstitium after 7 days. However, after 14 days, the 50 mg/kg dose of SCS was associated with vacuolization and loosening of germinal epithelium. These mild-to-moderate histopathological aberrations was confirmed from morphometric analysis in this dose group in which a decrease in the seminiferous tubules’ diameter and reduction in the thickness of germinal epithelium along with an increase in the interstitial area were observed. Conclusion: These findings concluded that SCS is considered to be relatively safe.

Impact of captive conditions on female germinal epithelium of the butterflyfish Chaetodon striatus (Perciformes: Chaetodontidae)

Summary Chaetodon striatus is a cosmopolitan seawater species present in aquaria all over the world and its extractivism is quite high. The lack of studies on the reproductive biology of C. striatus contributes to the difficulty in managing the species outside its natural habitat. Without knowledge of the mechanisms that control or affect gonadal changes, reproduction of C. striatus in captivity has become almost impossible, considering that the species is quite sensitive and the effect of captive conditions on its reproductive biology is unknown. Therefore, this study aimed to evaluate the effect on its reproductive biology of the animal’s confinement and possible alteration in structure of the ovaries. In C. striatus, after oocyte development, for animals confined in small spaces, maturing oocytes undergo atresia. During atresia, ovarian follicles were at different stages of degeneration, characterized by the progressive loss of the basement membrane and disorganization of the follicle complex. In the advanced stage of follicular atresia, there was total loss of the basement membrane, culminating in degradation of the follicle complex. In unconfined animals, oocyte development and maturation were not affected. Confinement also affected the cell structure of the germinal epithelium, which showed large numbers of apoptotic bodies. The difference in cortisol and glucose levels between the unconfined and confined groups was significant, which may have to do with the change found in the ovaries, such as extensive follicular atresia and loss of the basement membrane.

Histopathological changes due to arsenic Kushta in testes of wistar rats.

Objectives: To see the detrimental effects of arsenic kushta (kushta-e-Sammulfur) induced toxicity in wistar rats. Study Design: Experimental Study. Setting: Department of Morbid Anatomy and Histopathology, University of Health Sciences, Lahore. Period: 02nd June 2016 to 30th September 2016. Material & Methods: This experimental study was conducted on a total of 48 wistar rats, each weighing approximately, 200-300grams. These rats were then randomly divided into four groups each of them comprising of 12 rats. Group I was taken as a control group which was given flour pellets. Group II was given a single dose of 180 mg/kg of arsenic kushta for 2 weeks, whereas Groups III was given 150 mg/kg of arsenic kushta only for 12 weeks and Group IV was also given dose of 150 mg/kg of arsenic kushta along with 75mg of Bovine Serum Albumin for 12 weeks. Histopathological changes were then seen in the germinal epithelium, stages of spermatogenesis and insterstitium of testes of rats. Results: Changes in germinal epithelium, stages of spermatogenesis and interstitium were seen in all the above groups except group 1 which was the control group. Maximum changes were seen in groups C and D which were given high doses of arsenic kushta along with injection of bovine serum albumin. Conclusion: Arsenic kushta preparation of kushta-e-Summulfur causes testicular toxicity in wistar rats and have similar toxic effects in human beings.

The Effect of Propranolol on Sperm Parameters, CatSper 2 Gene and Protein Expression, and Oxidative Stress in Adult Mice

The aim of this study was to investigate the effects of propranolol on sperms, histopathology of testes, and CatSper 2 gene and protein expression in adult mice. 18 adult male mice were randomly divided into control, propranolol 1 (receiving 10 mg/kg dose) and propranolol 2 group (receiving 15 mg/kg dose for 35 days). The mean amount of sperm parameters in the propranolol 1 and propranolol 2 groups was significantly lower than the control group (p<0.05). CatSper2 gene and protein expression have significantly decreased in propranolol 1 and propranolol 2 groups compared to the control group (P<0.05). Reduction of CatSper2 gene and protein expression in low dose of propranolol was more severe than high dose. In testicular tissues of the propranolol 1 group, vacuoles and necrosis in the germinal epithelium were observed, and in testicular tissues of propranolol 2 group decrease in the thickness of the germinal epithelium, some vacuoles and necrosis were observed in germinal epithelium as well as congestion in the interstitial space. The mean value of thiol and catalase enzyme in the propranolol 1 and propranolol 2 groups, and the mean value of superoxide dismutase in propranolol 1 group, were significantly different compared to the control group (P<0.05).

Cellular development of the germinal epithelium during the female and male gametogenesis of Chaetodon striatus (Perciformes: Chaetodontidae)

SummaryButterflyfish Chaetodon striatus is highly sought after in the marine ornamental aquarium, although studies about its reproductive biology are scarce. Therefore, to contribute to a better understanding of the reproductive aspects of C. striatus, we describe in detail with the use of high resolution histology the cellular dynamics of the germinal epithelium during the reproductive life history of this species. Based on the activity of the germinal epithelium, this study describes different stages of the gonadal development, similar to the reproductive phases found in other fish, to determine the reproductive period of C. striatus. In characterization of gonadal development, the following germ cells are described for males: spermatogonia, spermatocytes, spermatids and spermatozoa. Oogonia, early, primary, secondary, full-grown and maturing oocytes are described for females. Female germinal epithelium of C. striatus showed substantial changes over the study period, indicating that there was an active spawning period. Male germinal epithelium also presented relevant alterations, indicating reproductive activity in the testicular lobules. Morphological data confirm how informative was the cellular dynamics of the germinal epithelium for understanding gonadal development during adult reproductive life of fish in general. Although Chaetodon are a popular species, previous studies have only produced superficial and rough histological analyses. Therefore, this study demonstrates important information on germinal epithelium of Chaetodon. This knowledge could be a fundamental tool for development of new strategies for breeding of several species in captivity, especially butterflyfishes.

Protective effects of vitamin E on aluminium sulphate-induced testicular damage

Male infertility can be caused by environmental factors, genetic defects, physiological and endocrine deficiencies and testicular pathologies. Aluminium (Al) can cause male infertility through a number of mechanisms. The aim of our study was thus to determine whether vitamin E (VitE) has protective effects on Al-induced testicular damage, which was determined according to sperm counts and morphology and using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method. Thirty-four male Wistar rats (250–300 g) were randomly assigned to control (no procedures performed; n = 6) or 0.2 mL intraperitoneal injection group ( n = 7 each; three times per week for 4 weeks): sham (distilled water), 10 mg/kg Al, 500 mg/kg VitE and 10 mg/kg Al plus 500 mg/kg VitE (Al + VitE). Sperm samples were evaluated for andrological parameters. The testes were examined by haematoxylin/eosin. The epithelial thickness and areas were calculated and Johnsen scores were determined for the germinal epithelium; the apoptotic indices were determined from TUNEL staining. For Al, the bonds between the germinal epithelial cells were broken in some tubules, and there were unidentified cells in the lumen of some tubules. For control, sham and VitE, normal morphology of the germinal epithelium was generally preserved. With Al + VitE, the full germinal epithelium cell series was maintained, with only mature sperm in the lumen. TUNEL-positive cells were significantly higher with Al compared to control and sham ( p < 0.05). For Al + VitE, the number of apoptotic cells was reduced compared to Al alone and was therefore similar to control, sham and VitE ( p > 0.05). Our findings show that Al caused testicular damage. VitE reduced the number of apoptotic cells during the damage caused by Al.