The testicular deficiency, cryptorchidism, varicocele, endocrine pathologies are the main causes of men infertility. The internal diseases of testicles lead to fertility declining in the major part of cases which explains the importance of studying of gonadopathies᾿ methods of correction. In recent times, vitamin D is noted among remedies suitable for effective correction of reproductive system. The aim of investigation: to determine the sex hormones levels and biochemical indices in serum of rats’ males with experimental testicles damage under the condition of vitamin D3 correction used separately or in combination with Tribestan®. Materials and methods. The experiment has been carried out on mature Vistar rats’ males; group of animals: S-model (animals with serotonin-induced gonads damage, gonadopathy (GP)); S+solvent (rats with gonadopathy which received solvent); S+vit. D3 (animals with gonadopathy received vitamin D3); S+Tr (rats with gonadopathy obtained Tribestan (Tr) – reference medicine); S+vit. D3+Tr (animals received both remedies); control – intact males. The activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), general content of nitrogen oxide cycle metabolites (NОх), concentration of TBARS and sex hormones Ts and E2 have been determined; their ratio has been calculated. Results. The NOх was increased in S-model rats. The application of vit. D3 or Tr has caused decreasing of NOх concentration in comparison with S-model group. The complex using of vit. D3 and Tr has had positive effect as for protection against oxidative stress (OS). The concentration of NOx in rats of S+vit. D3+Tr was by 56% less than in S-model group, but, as in the other experimental groups of animals, has remained increased relative to control group. The deficiency of free arginine in blood serum has been detected in all experimental groups comparing with Control group. The application of vit. D3 hasn’t affected on the arginine content. Tr has positively influenced the arginine level: the increasing of concentration of this amino acid was 95% relative to S-model group, but was less by 48% than values of intact animals. After application of Tr and vit. D3 together the concentration of arginine was higher by 2.2 times than in rats of S-model group, but hasn’t reached the values of intact animals. The concentration of TBARS has increased under the condition of GP. The using of vit. D3 has led to reducing, but not to normalizing of TBARS content. The concentration of TBARS in rats of S+Tr group was by 1.9 times lower than in rats with GP and by two times higher than in intact animals. Simultaneous using of Tr and vit. D3 has been effective as for reducing of lipid peroxidation (LP) intensity. The concentration of TBARS in S+vit. D3+Tr group has declined by 2.2 times comparing with values of animals with GP. The combination of Tr and vitamin D3 was more effective than cholecalciferol single using as for protection against OS and LP reducing and has demonstrated activity at the level of reference drug. GP has led to increasing of ALT activity 2.4 times and AST – by 2 times comparing with control group. The application of Tr, vit. D3 or their complex hasn’t changed ALT activity which was statistically higher than intact animals’ values. The activity of AST was increased in animals with GP and reduced after experimental correction of GP consequences. Vitamin D3 monotherapy has reduced by 32% the activity of AST comparing with animals with GP. AST concentration in males’ blood serum was decreased too with the combined use of Tr and vitamin D3 comparing with untreated animals. Thus, vitamin D has positively impacted the AST level in combined use with Tr as well as monotherapy. Serotonin-induced experimental GP has caused the declining of general Ts concentration in blood serum of animals of all experimental groups. The application of vitamin D3 hasn’t demonstrated androgen correcting activity. Tr has positively influenced androgen synthesized function: Ts concentration has increased by 95% comparing with S-model group, but remained lower by 31% than in intact animals. The Ts concentration in S+vit. D3+Tr group was 2.1 times higher than in animals with GP. However, Ts values in this group haven’t reached Control group values and were by 25% less. The concentration of estradiol in all experimental groups has corresponded to the values of intact animals. The calculation of Ts/Е2 ratio has shown to be decreased in S-model, S+solvent, S+vit. D3 groups in comparison with intact animals. The Ts/Е2 ratio has normalized in groups with GP treated by reference drug separately or in combination with vitamin D3. Conclusions: The application of cholecalciferol has considerably decreased NOx and TBARS concentrations, but hasn’t affected on arginine level which has declined with the gonadopathy. Vitamin D has caused reducing of aspartate aminotransferase activity in serum and hasn’t had androgenic and estrogenic effects. The application of Tribestan has led to the pro-oxidative/antioxidative balance recovery under the condition of experimental gonadopathy. This medicine has reduced the high level of aspartate aminotransferase activity, increased testosterone level and normalized testosterone/estradiol ratio in blood serum of experimental animals. The application of vitamin D and Tribestan in combination has led to improvement of androgenic status and has demonstrated better effect as for protection against oxidative stress and lipid peroxidation than cholecalciferol as a monotherapy.
Actuality. Nowadays, the disturbances of men’s reproductive function are the important problem which often accompany sex glands diseases. During last years, scientific researchers are taking into consideration vitamin D with aim to create safe medicines or treatment schemes for therapy hypofertility developed due to gonadopathies. It is thought, Vitamin D may improve not only spermogramm, but, probably, acts on men’s reproductive ability by influencing fertility. The correction of reproductive disturbances requires the experimental grounding of vitamin D and its complex schemes using on the infertility experimental models. The aim of this investigation was to determine the condition of rat males’ reproductive system with experimental damage of testicles during correction by vitamin D3. Materials and methods. The correction by vitamin D3 (cholecalciferol) as a monotherapy or together with Tribestan® has been carried out in Vistar rats with experimental gonadopathy induced by Serotonin hydrochloride. The drug of comparison Tribestan® has been introduced per os in dose of 68 mg/kg body mass three days before Serotonin hydrochloride introduction; under the condition of serotonin obtaining (14 days) and during three days after its last injection once a day. Vitamin D3 has been given according to the same scheme in value of 0,5 ml in dose of 4000 IU (per os). The male fertility has been determined according to fertility and fecundity indices of rats’ females fertilized by experimental rats’ males. Due to results of mating with intact females the integral index of average realized fecundity (fertility) of rats’ males has been calculated. The researches of spermatogenesis has been carried out in epididymis’s suspension, the morphometric indices were studied on the histological sections. Results. Under the condition of serotonin induced gonadopathy the disturbance of all indices of fertility and reproductive function has been observed. The introduction of cholecalciferol as a monotherapy during three weeks has statistically increased the part of motile epididymal sperms relative to gonadopathy, but not reaching the intact animals level; the number of spermatogonia and index of spermatogenesis have been normalized. However, the vitamin D using hasn’t influenced general number and percentage of pathological forms of sperm cells in rats with modelled testicles damage; hasn’t improve males’ fertility. The introduction of vitamin D3 together with Tribestan to rats mails with serotonin induced testicles damage has positively affected all the spermogramm and gonads morphometric indices. The fecundity, as an embryonic losses level, has almost declined to Control rats’ values, whereas, the number of alive fetuses in thus group has been statistically higher than in groups with pathology as well as in those animals which obtained only cholecalciferol or referent medicine separately. The index of realized fecundity Fi has normalized in rats obtained vitamin D3 together with Tribestan. Fi in this group was higher than in other experimental animals. Conclusions. The using of cholecalciferol in complex schemes of treating of reproductive system disturbances due to gonads damage has led to improving of reproductive function and may be considered being pathogenetic therapy due to influence the different chains of reproductive health.
It was studied parameters of sexual activity and quality of spermatogenesis in male rats at the age of 4–4.5, 10, 18 and 20–21 months. It was found that the age involution of sexual activity manifested by changes in the qualitative composition of the group initially sexually active males (decrease in the proportion of animals that are capable of ejaculation and the beginning of the next series of couples), as well as changing the sexual behavior pattern (strengthening of court behavior, weakening of the copulatory behavior and slowing of the postejaculatory recovery). It was identified nonlinear character of age changes in spermatogenesis as the best semen parameters in animals 10 months of age compared with younger and older males. It was suggested that asynchrony of the age involution of the different components of the male reproductive function has adaptive sense.
One of the current problems is the study of the effects of vitamin D on the body, and in men its action is closely related to the pathogenesis of androgen deficiency and hypofertility. Particular attention needs to be paid to determining whether cholecalciferol (D3) has a negative effect on the gonads and spermatogenesis of intact individuals, as vitamin D therapy is used in reproductive disorders with or without vitamin D deficiency. The aim of the study was to determine the effect of vitamin D on the histological structure of gonads and morphometric parameters of spermatogenesis of adult intact male rats. The studies were performed on adult sexually active male Wistar rats. Vitamin D3 was administered orally in doses of 1000 IU, 4000 IU and 10000 IU. The solutions were made on seed oil. The control was intact rats. Vitamin D and its solvent were administered throughout the period of spermatogenesis and the time of maturation of sperm in the epididymis, after which the structural organization of the testes was determined. Gonadal samples were fixed in 10 % formalin solution, leave in alcohols of increasing strength, and embedded in paraffin. In addition to survey microscopy, morphometric evaluation of the process of spermatogenesis was performed on sections of gonads stained with hematoxylin and eosin. Micropreparations were examined using a Granum L 30 (03) light microscope, and microscopic images were taken with a Granum DSM 310 digital video camera. Photographs were processed on a Pentium 2.4 GHz computer using Toup View. Statistical processing of data results was performed in the standard software package "Statistica 6.0" using Student's t-test and using a non-parametric analogue of one-way analysis of variance – Kruskal-Wallis criterion. In rat testicular sections, seminiferous tubules were located transversely or obliquely and were oval or round in shape. The diameter of the tubules is normal, the tubular membrane, as well as the protein and vascular membranes were normal. The basal department contains the youngest cells of the germinal epithelium – spermatogonia. Cells have a pronounced functional activity. Morphometric characteristics of spermatogenesis of intact rats corresponded to the physiological norm. The introduction of the solvent throughout the period of spermatogenesis and maturation of mature sperm in the epididymis did not affect the histoarchitectonics of the testicles. The testicular lobes are filled with concentric or flattened profiles of sections of the seminal tubules, which are close enough to each other. The diameter of the tubules is normal, the intrinsic membrane of the tubules, as well as the protein and vascular membranes corresponded to those in intact animals. 3-4 generations of spermatogenic cells, which were at different stages of development, can be seen in the tubules. However, few tubules with focal destruction of the germinal epithelium and exfoliation of germ cells in the lumen of the tubules have been observed. No significant changes in the microstructure of the seminal tubules were observed after administration of vitamin D at all doses studied. Not only spermatogenesis but also spermiogenesis is clearly traced in different tubules of rats – stages of cellular transformations from spermatid to sperm. Morphometric parameters of the process of spermatogenesis of rats receiving different doses of vitamin D3 do not differ from those of intact rats. Thus, the use of vitamin D in these doses revealed the safety of its effect on the number of spermatogonia and tubules with stage 12 meiosis. When cholecaciferol was used in male rats for 68 days, the spermatogenesis index remained at the level of animals that did not receive the test substance. The use of vitamin D3 in intact adult male rats does not adversely affect the histological structure of the testes.
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