Effect of [R]-[+] pulegone on ovarian tissue; correlation with expression of aromatase Cyp19 and ovarian selected genes in mice

Objective: Pulegone [PGN] is a monoterpene ketone, whose metabolites exert several cytotoxic effects in various tissues. The present study was conducted in order to evaluate the [R]-[+] PGN-induced alterations in ovarian aromatization, proto-oncogenes and estrogen receptoralpha [ER alpha] and ER beta receptors expressions

Materials and Methods: In this experimental study, mature albino mice were divided into experimental [received 25 mg/kg, 50 mg/kg and 100 mg/kg PGN, orally for 35 days] and control [received 2% solution of Tween 80 as a PGN solvent, orally] groups. The mRNA levels of Er alpha, Er beta, p53, Bcl-2, and cytochrome p450 [Cyp19] as well as ovarian angiogenesis were analyzed through reverse transcription polymerase chain reaction and immunohistochemical techniques, respectively. Moreover, apoptosis of follicular cells, serum estrogen and progesterone levels and mRNA damage were investigated via using terminal transferase and biotin-16-dUTP staining, electrochemilunescence and fluorescent microscopy methods, respectively

Results: The PGN reduced Er alpha, Er beta and Cyp19 expression at 50 mg/kg and 100 mg/kg doses, while significantly elevating p53 and reducing Bcl-2 expression. Finally, PGN impaired ovarian angiogenesis, increased apoptosis, elevated follicular atresia and reduced serum levels of estrogen and progesterone

Conclusion: Chronic exposure to PGN [50 mg/kg and 100 mg/kg], severely affects ovarian aromatization, proto- oncogenes mRNA levels and expression of ERs

Co-administration of vitamin E and testosterone attenuates the atrazine-induced toxic effects on sperm quality and testes in rats

Objective: Atrazine [ATZ] as a widely used herbicide is considered as a potent endocrine disrupter which adversely affects reproductive systems in both genders. This study aimed to assess the effects of testosterone [T]- and vitamin E [VitE]- alone and their co-administration on testicular function and sperm parameters after exposure to ATZ in rats

Materials and Methods: In this experimental study, the rats [n=30] are assigned into the following 5 groups: control-sham group [n=6] receiving corn oil, ATZ group [n=6] receiving 200 mg/kg ATZ alone, ATZ+VitE group [n=6] receiving 150 mg/kg ATZ+VitE, ATZ+T group [n=6] receiving 400 micro g/kg ATZ+T, and ATZ+VitE+T group [n=6] receiving ATZ+VitE+T for 48 consecutive days. Total antioxidant capacity [TAC], total thiol molecules [TTM], and malondialdehyde [MDA] were analyzed. Serum levels of T, luteinizing hormone [LH], and inhibin-B [IN-B] were also determined. Histological examination and sperm analysis were performed. The data were analyzed using Graph-Pad Prism software version 2.01

Results: Co-administration of VitE and T significantly [P<0.05] increased ATZ-decreased TAC and TTM levels and reduced ATZ-increased MDA content. T and VitE significantly [P<0.05] increased serum levels of ATZ-reduced T [1.94 +/- 0.96], IN-B [122.10 +/- 24.33] and LH [0.40 +/- 0.10]. The T+VitE animals showed a reduction in apoptotic cells and an increase in Leydig cells steroidogenesis. Co-administration of T and VitE significantly [P<0.05] reduced the ATZ-induced DNA disintegrity and chromatin de-condensation. VitE and T protected germinal cells RNA and protein contents against ATZ-induced damages

Conclusion: T and VitE in simultaneous form of administration were able to normalize the ATZ-induced derangements through promoting antioxidant capacity and endocrine function

Effect of phosalone on testicular tissue and in vitro fertilizing potential

The current study aimed to evaluate the effects of phosalone [PLN] as an organophosphate [OP] compound on testicular tissue, hormonal alterations and embryo development in rats. In this experimental study, we divided 18 mature Wistar rats into three groups-control, control-sham and test [n=6 per group]. Animals in the test group received one-fourth the lethal dose [LD50] of PLN [150 mg/kg], orally, once per day for 45 days. DNA laddering and epi-fluorescent analyses were performed to evaluate testicular DNA fragmentation and RNA damage, respectively. Serum levels of testosterone and inhibin-B [IN-B] were evaluated. Testicular levels of total antioxidant capacity [TAC], total thiol molecules [TTM] and glutathione peroxidase [GSH-px] were analyzed. Finally, we estimated sperm parameters and effect of PLN on embryo development. Two-way ANOVA was used for statistical analyses. There was severe DNA fragmentation and RNA damage in testicular tissue of animals that received PLN. PLN remarkably [p<0.05] decreased testicular TAC, TTM and GSH-px levels. Animals that received PLN exhibited significantly [p<0.05] decreased serum levels of testosterone and IN-B. Reduced sperm count, viability, motility, chromatin condensation and elevated sperm DNA damage were observed in the test group rats. PLN resulted in significant [p<0.05] reduction of in vitro fertilizing [IVF] potential and elevated embryonic degeneration. PLN reduced fertilization potential and embryo development were attributed to a cascade of impacts on the testicles and sperm. PLN promoted its impact by elevating DNA and RNA damages via down-regulation of testicular endocrine activity and antioxidant status

Sulpiride-induced hyperprolactinemia in mature female rats: evidence for alterations in the reproductive system, pituitary and ovarian hormones

Background: The prevalence of hyperprolactinemia following administration of conventional antipsychotic drugs requires further investigation. The current study is designed to evaluate the effect of sulpiride [SPD]-induced hyperprolactinemia on alterations to ovarian follicular growth, gonadotropins, and ovarian hormones and to analyze the extent of potential problems in mammary glands

Materials and Methods: A total of 40 albino Wistar rats were divided into four groups: control [no treatment], control-sham [0.3 ml olive oil], low dose SPD [20 mg/kg] and high dose SPD [40 mg/kg]. All compounds were intraperitoneally [IP] administered for a period of 28 days

Results: After 28 days, we dissected the rats' ovarian tissues, uterine horns and mammary glands which were sent for histological analyses. We counted the numbers of normal, atretic follicles and corpora lutea [CL]. Serum levels of prolactin [PRL], estradiol, progesterone, follicle stimulating hormone [FSH] and luteinizing hormone [LH] were evaluated. SPD-administered animals showed sporadic follicular atresia in different sizes associated with higher numbers of CL on the ovaries. The mammary glands exhibited features of galactorrhea. There was remarkable [p<0.05] elevation in SPD-administered animals' uterine horn endometrium, myometrium and perimetrium thicknesses. The serum levels of PRL and progesterone significantly [p<0.05] increased, while the serum concentration of estradiol, LH and FSH notably [p<0.05] decreased according to the SPD administered dose. No histological and biological changes occurred in control-sham animals. SPD-induced animals had unsuccessful attempts at mating and decreased pregnancy rates

Conclusion: The present findings suggest that SPD-induced disturbances depend on PRL level. In addition, an increased PRL level is largely dependent on the administered doses of SPD

[ effects of vitamin E and exogenous testosterone co-administration on biochemical serum parameters of varicocele rats]

Because of impairment of testosterone production and occurrence of oxidant disorders in testis of varicocele [VCL] patients, these patients are treated with testosterone and various antioxidant. This study was performed with the objective of determining the effect of co-administration of these two drugs on the serum biochemical parameters of rats. In this study, 30 male rats were divided into 5 groups [n=6, each] of control-sham and test groups. In the control-sham group, only simple laparotomy was performed. In the test groups, experimental VCL was induced in all test groups. No treatment was performed for the first test group. The second test group was treated with testosterone [44microg/kg, ip], the third group with vitamin E [150mg/kg, orally] and the fourth group with co-administration of testosterone and vitamin E. After the end of treatment period, anesthesia and heart blood sampling were performed, and after serum separation, serum levels of glucose, creatinine, urea, total cholesterol, and triglyceride were measured. The data were analyzed using two-way variance analysis. The significance level was considered to be p < 0.05. In this study, the serum levels of the glucose, creatinine, urea, total cholesterol, and triglyceride significantly increased following VCL induction, as compared to control group [p < 0.05]. This was while single administration of testosterone and co-administration with vitamin E significantly reduced the serum levels of mentioned parameters [p < 0.05]. The results showed that co-administration of testosterone and vitamin E, in comparison with separate administration of each compound, improved the serum biochemical parameters in VCL-induced animals

[ effect of vitamin E and dexamethasone co-administration on varicocle-induced biochemical and histological alterations of liver in rats]

Varicocele [VCL] not only has functional and structural effects on the male reproductive system, but also affects other body tissues such as liver. Therefore, dexamethasone [DEX] and vitamin E [VE] co-administration is recommended invaricocele patients. The present study was conducted with the purpose of evaluating the effect of experimentally-induced varicocele on liver and the protective effects of VE and DEX. In this study, 30 male rats were divided into five groups. The experimental group were assigned to 4 groups: Varicocele; varicocele+VE [150/kg]; varicocele+DEX [0.25mg/kg]; and varicocele+VE+DEX. The control group underwent a simple laparotomy. After 60 days, the rats were anesthetized and blood was taken for serum isolation. Simultaneously with taking blood samples,the serum levels of glucose, alanine aminotransferase [ALT], and aspartate aminotransferase[AST] were evaluated. The hepatocytes' cytoplasmic carbohydrate level was assessed by periodic acid shiff staining. Statistical analysis was performed by analysis of variance and Tukey's test. The significance level was considered as p<0.05. The levels of blood glucose, ALT, and AST were significant lyreduced in VE and DEX groups compared to varicocele group. Also, tissue necrosis, central venous dilatation, and lymphatic cells infiltration decreased. According to the findings of this study, varicocelecauses liver damage at the biochemical and histological levels, and co-administration of VE and DEX could significantly reduce the VCL-induced complications and also DEX-induced side effects as well

Paraquat exposure up-regulates cyclooxygenase-2 in the lungs, liver and kidneys in rats

Paraquat is a commonly used herbicide in many countries which can lead to systematic poisoning on exposure, In this study, paraquat [PQ]-induced changes in the expression of Cyclooxygenase-2 [COX-2] along with biochemical and histopathological changes in the lungs, liver and kidneys were studied. Twenty four male Wistar rats [180-200 g] were exposed either against saline as control group or various doses of PQ [3.5, 7 and 10 mg/kg, SC] as test groups for 7 consecutive days. The animals in test groups demonstrated a significant increase of malondialdehyde and NO contents, while a remarkable decrease of total thiol molecules was recorded. Histopathological studies revealed a severe alveolar edema and hemorrhages in the lungs, congestion and glycogen degeneration in the liver and multifocal interstitial nephritis along with proximal tubular degeneration in the kidneys. PQ up-regulated the COX-2 expression at mRNA level significantly in the examined organs. This data suggest that the PQ-induced oxidative disturbances and pathological damages can be attributed to the upregulation of COX-2 in examined organs

Honey and metformin ameliorated diabetes-induced damages in testis of rat; correlation with hormonal changes

The global prevalence of diabetes mellitus is on rise. Diabetes-induced oxidative stress has been known to affect liver, pancreas, kidney and reproductive organs pathologically. Honey is a natural product of bee with antioxidant properties. Current study aimed to analyze the protective effects of Metformin [MF] alone and MF+ natural honey co-administration on diabetes-induced histological derangements in testis of rats. Thirty six, mature male Wistar rats were randomly divided into six groups including; control, honey-dosed non-diabetic, diabetes-induced [65 mg/kg, single dose], honey-administrated diabetic [1.0 g/kg/day], Metformin-received diabetic [100 mg/kg/day], Metformin and honey-co-treated diabetic which were followed 40 days. The animals were anesthetized by diethyl ether and the blood samples were collected. The serum levels of testosterone, Insulin, LH and FSH analyzed using antibody enzyme immunoassay method. The testicular tissues were dissected out and underwent to histological analyses. The biochemical analyses revealed that the diabetes resulted in significantly reduced testosterone [p<0.01], LH and FSH [P<0.01, 0.001] levels in serum. Light microscopic analyses showed remarkable [p<0.01] reduction in seminiferous tubules diameter [STD], spermiogenesis index [SPI] and thickness of the epithelium in the diabetic group versus control and co-treated groups. Simultaneous administration of the honey with MF could fairly up-regulate testosterone, LH and FSH levels. The animals in metformin and honey-treated group exhibited with improved tubules atrophy, elevated spermiogenesis index and germinal epithelium thickness. Our data indicated that co-administration of Metformin and honey could inhibit the diabetes-induced damages in testicular tissue. Moreover, the simultaneous administration of metformin and honey up-regulated the diabetes-reduced insulin, LH, FSH and testosterone levels

Uterine artery interruption: evidence for follicular growth and histochemical and biochemical changes

The present study was conducted in order to evaluate the effects of bilateral uterine artery ligation [BUAL] on the ovarian follicular fate, and alterations in carbohydrate, lipid, lipase and serum levels of F9SH, LH, prolactin, estrogen and progesterone. Twenty-four mature female rabbits divided into two test and control-sham groups. The animals underwent ovariohystrectomy on days 23, 43 and 63 after BUAL. Later serum and tissue samples were processed for histological and biochemical analyses. Two-way ANOVA test was used for statistical analyses and p<0.05 was considered as significant. The ovaries from the case groups exhibited markedly increased atretic follicles, which were characterized by early antrum formation, ooplasmic vacoulation, granulosa cells dissociation and oocyte deformation. Lipid foci were remarkably present in the cytoplasm of oocytes, granulosa and theca cells in BUAL rabbits. Smaller sized atretic follicles showed higher lipid reactions than large ones. The PAS reaction was highly positive in zona pellucida [ZP], basement membrane, granulosa cells and follicular fluid of atretic follicles. Early atresiated follicles showed remarkable reaction sites for lipase. Significant [p<0.05] increase in serum levels of FSH, LH, progesterone, and prolactin was revealed in BUAL rabbits compared to the control group while serum levels of estrogen decreased time-dependently in the test groups. The current study suggests the critical role of the uterine artery in controlling ovulation and follicular growth. Moreover atresia processes might relate to lipid accumulation in the cells along with attenuation of lipase activity

Histological and histochemical effects of Gly-phosate on testicular tissue and function

In this study we aimed to evaluate the impact of chronic exposure to the Gly-phosate [GP] on rat's testicular tissue and sperm parameters. Testicular tissue, morphology of sperms and testosterone level in serum of mature male rats were analyzed. Animals were divided into two test and control-sham groups. The test group was subdivided into 4 groups [10, 20, 30 and 40 days GP administrated]. Each test group [n=8] received the compound at dose of 125 mg/kg, once a day, orally for 40 days while control-sham group [n=16] received the corn oil [0.2 ml/day]. Microscopic analyses revealed increased thickness of tunica albuginea, obvious edema in sub-capsular and interstitial connective tissue, atrophied seminiferous tubules, arrested spermatogenesis, negative tubular differentiation and repopulation indexes, decreased Leydig cells/mm[2] of interstitial tissue, hypertrophy and cytoplasmic granulation of Leydig cells, elevated death, immature sperm and increased immotile and abnormal sperm percentage. The carbohydrate ratio was reduced in first three layers of the germinal epithelium [GE] cytoplasm. The upper layers of the GE series were manifested with low rate of lipid accumulation in cytoplasm, while the cells which were located in first layers were revealed with higher amount of lipid foci. Hematological investigations showed significant [p<0.05] decreasing of testosterone level in serum. The current data provide inclusive histological feature of chronic exposure against GP with emphasizing on reproductive disorders including histological adverse effect on the testicular tissue, spermatogenesis, sperm viability and abnormality which potentially can cause infertility

Testicular biohistochemical alterations following experimental varicocele in rats

The exact pathophysiology of testicular degeneration, following varicocele has not been completely understood yet. The current study was designed to determine the effect of varicocele on germinal epithelium [GE] cytoplasmic biohistochmical alterations. To follow-up this study, left varicocele was induced in test groups. Non-varicocelized rats were served as control-sham [n=6]. Following 4, 6 and 8 months, right and left testes were dissected out and the blood serum sample was taken. The GE cytoplasmic carbohydrate, lipid accumulation, lipase and alkaline-phosphates [ALP] ratios were analyzed. Serum levels of LH, FSH and testosterone were measured. Observations demonstrated that in varicocele-induced rats, the spermatogenesis cell lineage exhibited lower number of cells with periodic acid shift positive cytoplasm, higher number of cells with lipid and ALP positive stained cytoplasm in comparison to control animals. Lipase enzyme decreased by the time in the test animals. In varicocelized groups the number of Leydig cells decreased in to 2.25 +/- 0.41 and 1.16 +/- 0.75 per one mm[2] in left and right testicles respectively after 8 months, and these cells demonstrated an ALP positive feature. In test groups, the serum levels of LH and FSH reduced into 1.12 +/- 0.01 and 2.03 +/- 0.05 ng/ml respectively after 8 months. Although testosterone level diminished by the time in the test animals, and this decreasing was significant [p=0.031] after 8 months [3.08 +/- 0.10 ng/ml]. Our results suggest that following varicocele induction major alterations occur in GE, which may lead to loss of GE cells physiological function and ultimately result in fertility problems

Varicocele time-dependently affects DNA integrity of sperm cells: evidence for lower in vitro fertilization rate in varicocele-positive rats

We designed this study to clarify how varicocele can time-dependently affect sperm morphological parameters and DNA integrity. In this study, we intend to estimate the effect of various periods of varicocele on the in vitro fertilization [IVF] rate in rats. In this experimental study, left varicocele were induced as the test group [n=18] which was further sub-divided into three groups based on the study termination time [4, 6 and 8 months after varicocele induction]. The control-sham group [n=6] consisted of rats who received no treatment. Repopulation index [RI], tubular differentiation index [TDI], sperm viability and motility, morphological maturity, chromatin integrity and ability to undergo IVF were assessed. In addition, the potential impact of varicocele on serum total antioxidant capacity [TAOC] and total thiol molecules [TTM] were examined. Histological results showed that varicocele negatively influenced TDI and RI. All sperm morphological parameters were lower than those in the control-sham group. DNA damage was severely and time-dependently substantiated in all test groups. Varicocele significantly reduced the ability of sperm derived from varicocele rats to undergo IVF. Serum TAOC and TTM levels reduced in a time-dependent manner. Right testes varicocele-induced rats showed remarkably less damaged profile for all investigated parameters compared to the left testes varicocele. Our data suggested that experimentally induced varicocele negatively impacted sperm maturation and chromatin integrity in a time-dependent manner. This consequently caused a remarkable reduction in IVF ability. The detrimental effect of varicocele may be attributed to the significant reduction of antioxidant capacity of the serum

Follicular viability and histological alterations after auto-transplantation of dog ovaries by experimentally inducing blood sinus on stomach

Currently, chemotherapy and radiotherapy and considered most effective methods for cancer treatment, however these strategies often result in fertility problems. A favorable alternative to prevent fertility loss in cancer patients is the cryopreservation and transplantation of sexual tissues [ovaries and/or testes]. There is a low rate of fertilization following cryopreservation of ovaries prior to implantation. Therefore, in our opinion, this low rate is caused by instable blood flow during organ transplantation. Thus, this study researches a canine ovarian model that focuses on direct exposure of ovaries with blood in an experimentally induced sinus-like cavity. We implanted this tissue on the muscular layer of the stomach, which is its most vascularized region. Ovarian transplantation was conducted on T1 animals [n=5], bilateral ovariectomy was performed on T2 animals [n=5], unilateral ovariectomy was conducted on T3 cases and animals in the control-sham group [n=5] did not undergo ovariectomy or transplantation. All isotransplanted ovaries survived. Ovaries resumed follicular growth and revascularization. Transplanted ovaries contained 75%-76% of survived small follicles [pre antral] after 60 days. The ovarian granulosa cells showed considerable resistance against ischemia. After day 30 no statistically significant differences in the level of estradiol and progesterone were observed between T1 animals and the T3 group. T1 animals showed considerably high level of progesterone and estradiol in comparison to T2 cases. This study showed that using blood sinus method for ovarian isotransplantation helps ovarian tissue to survive from post implantation ischemia which confirms with normal follicles presentation and intact endocrine function of the implanted ovaries