Effects of lupeol and flutamide on experimentally-induced polycystic ovary syndrome in mice.

Objectives: Polycystic ovary syndrome (PCOS) is one of the main causes of infertility in women. This study was conducted to uncover the effects of lupeol as an anti-androgenic triterpene on experimentally-induced PCOS in mice. Materials and Methods: Eighty immature female mice were divided into 4 groups: Control (C), PCOS (P), Lupeol (L), and Flutamide (F). PCOS was induced in test groups by injection of Dehydroepiandrosterone (60 mg/kg/day, IP) for twenty days. Following the PCOS induction, the two groups of L and F were treated with lupeol (40 mg/kg/day) and/or flutamide (10 mg/kg/day) respectively and the two groups of C and P received sesame oil (0.1 ml/mouse/day) for 15 days. After the treatment period, ten animals in each group were selected for collecting blood and ovary samples. In vitro fertilization assessment was carried out on 10 remaining mice in each group. The hormonal assays and oxidative stress biomarker determination were performed on serum and tissue samples. Moreover, histopathological analyses were conducted on the ovaries. Results: PCOS-elevated concentration of LH and Testosterone was significantly (P<0.05) lowered in lupeol and flutamide-received animals. Lupeol and flutamide both reduced PCOS-induced fibrosis and the number of atretic follicles. Both compounds declined the PCOS-increased lipid peroxidation and protein oxidation in serum and the ovaries. Lupeol increased the PCOS-reduced fertility rate and decreased the number of arrested embryos by 12%. Conclusion: These findings indicate that lupeol could be a novel compound in the treatment of PCOS as it reduced PCOS-induced structural and also functional disorders.

Protective effects of Equisetum arvense methanolic extract on testicular tissue disorders in streptozotocin-induced diabetic murine model.

Diabetes in a long period can damage the testicular tissue and impair the male fertility potential. Recently, different herbal treatments have been used for the prevention of type I diabetes and its pathological effects. Methanolic extract of Equisetum arvense has anti-oxidant and hypoglycemic properties. Thus, the current study aimed to evaluate the protective effects of Equisetum arvense methanolic extract (EE) on diabetes-induced detrimental effects in mice testicular tissue. Thirty-two adult male mice were randomly divided into four groups including control-sham, diabetic (induced by streptozotocin, 50.00 mg kg-1 for five days), diabetic + EE 250 (250 mg kg-1) and diabetic + EE 500 (500 mg kg-1). After 45 days, all animals were euthanized and their testicles were dissected out and undergone histological analyses. Moreover, the serum level of testosterone was evaluated. Analyses showed that seminiferous tubules diameter, Leydig cells number per mm2 of the connective tissue, Sertoli cells number per tubule, serum level of testosterone and percentage of seminiferous tubules with positive tubular differentiation, repopulation and spermiogenesis indices were significantly decreased in the diabetic group in comparison with control-sham group. The administration of EE in test groups significantly decreased the adverse effects of diabetes (especially 500 mg kg-1). The results of this study revealed that diabetes disturbs spermatogenesis and spermiogenesis processes in mice. Meanwhile, the EE prevents diabetes-induced damages in mice testicular tissue, which may be associated with its hypoglycemic and antioxidative activities.

Protective effects of Equisetum arvense methanolic extract on sperm characteristics and in vitro fertilization potential in experimental diabetic mice: An experimental study.

BACKGROUND: Diabetes mellitus is a metabolic disorder characterized by impaired insulin secretion or the inability of tissues to respond to insulin. This disease can damage the testis and reduce semen quality. Therefore, it can impair the potential for male fertility. Different herbal therapeutic treatments have been used to control diabetes and its complications. OBJECTIVE: This study aimed to evaluate the effects of streptozotocin-induced diabetes on sperm and in vitro fertilization (IVF) potential and investigate the protective effects of Equisetum arvense methanolic extract on diabetic mice. MATERIALS AND METHODS: Twenty-four adult male mice were divided into four groups: control-sham, diabetic, diabetic + Equisetum extract (250 mg/kg), and diabetic + Equisetum extract (500 mg/kg). After 45 days, sperm samples were collected from the cauda epididymis to evaluate the characteristics of sperm (including viability, count, motility, morphology and chromatin/DNA integrity of sperm) and the IVF potential. RESULTS: Sperm motility and viability were increased remarkably (p ≤ 0.001) in the treated groups compared with the non-treated diabetic group. The decrease in sperm count in the diabetic group compared with the treated groups was not significant. Moreover, the percentage of sperm with DNA damage, nuclear immaturity, and abnormal morphology was decreased significantly (p ≤ 0.001) in the treated groups compared with the diabetic group. The treated animals exhibited remarkably higher fertilization rates and a higher percentage of fertilized oocytes that developed toward the blastocyst stage compared with the non-treated diabetic group (p ≤ 0.001). CONCLUSION: The methanolic extract of the Equisetum arvense inhibited diabetes-induced detrimental effects on sperm quality and fertilization rate, which may have been associated with hypoglycemic and antioxidative activities in this plant.

Silymarin and celecoxib ameliorate experimental varicocele-induced pathogenesis: evidences for oxidative stress and inflammation inhibition.

BACKGROUND: The present study was done to investigate the ameliorative effect of silymarin (SMN) and celecoxib (CEL) on varicocele (VCL)-induced detrimental impact in testicular tissue. METHODS: Mature Wistar rats were divided into control and test groups. Following VCL induction, the animals in test group were subdivided into non-treated VCL-induced, SMN-treated (50 mg/kg, orally), CEL-treated (10 mg/kg) and SMN + CEL-treated groups. Following 60 days, testicular total antioxidant capacity (TAC), malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), glutathione peroxidase (GSH-px), total thiol molecules (TTM), mRNA and protein levels of COX2 and mRNA level of iNos were analyzed. Moreover, the germinal cells apoptosis and mRNA damage were examined. RESULTS: Observations revealed that co-administration of SMN and CEL significantly (P < 0.05) up-regulated TAC, SOD, GSH-px and TTM levels and resulted in a remarkable (P < 0.05) reduction in iNos and COX2 expression, NO and MDA contents. The animals in SMN + CEL-treated group exhibited significantly (P < 0.05) lower number of apoptotic cells and cells with mRNA damage per one mm2. CONCLUSION: The SMN by up-regulating testicular TAC, SOD, GSH-px and TTM levels and the CEL by inhibiting COX2 and iNos expression as well as NO content could fairly ameliorate the VCL-decreased spermatogenesis.

Nanosilver particles increase follicular atresia: Correlation with oxidative stress and aromatization.

Present study was performed in order to update the possible mechanism(s), involving in nanosilver particles (NSPs)-induced detrimental impacts in ovarian tissue. For this purpose, 24 mature female rats were divided into control and 0.5, 1, 5 mg/kg NSPs-received groups (intraperitoneally, for 35 days). Follicular growth and atresia, ovarian total antioxidant capacity (TAC), malondialdehyde (MDA), superoxide dismutase (SOD) contents, serum estrogen (E2 ) level and macrophages infiltration were investigated. Moreover, ovarian angiogenesis, cellular mRNA damage and cytochrome aromatase CYP19 expression were analyzed. The NSPs enhanced follicular atresia diminished E2, reduced TAC and SOD level, elevated MDA content and up-regulated macrophages infiltration. Cellular mRNA damage, impaired angiogenesis and diminished CYP19 expression were revealed in NSPs-received groups. Therefore NSPs by down-regulating aromatization, reduce E2 synthesis which then it leads to impaired angiogenesis. The impaired angiogenesis in turn down-regulates ovarian antioxidant status, which partially enhances follicular atresia by triggering lipid peroxidation and mRNA damage.

PCOS women show significantly higher homocysteine level, independent to glucose and E2 level.

BACKGROUND: It is reasonable to think that some biochemical characteristics of follicular fluid (FF) surrounding the oocyte may play a critical role in determining the quality of oocyte and the subsequent potential needed to achieve fertilization and embryo development. OBJECTIVE: This study was carried out to evaluate the levels of FF homocysteine (Hcy) in IVF candidate polycystic ovary syndrome (PCOS) women and any relationships with FF glucose and estradiol (E2) levels. MATERIALS AND METHODS: In this case control study which was performed in Dr. Tizro Day Care and IVF Center 70 infertile patients were enrolled in two groups: comprising 35 PCOS and 35 non PCOS women. Long protocol was performed for all patients. FF Hcy, glucose and E2 levels were analyzed at the time of oocyte retrieval. RESULTS: It was observed that FF Hcy level was significantly higher in PCOS patients compared with non PCOSs (p<0.01). Observations demonstrated that in PCOS group, the Hcy level increased independent to E2, glucose levels, BMI and age, while the PCOS group showed significantly higher BMI compared with non-PCOS group (p=0.03). However, no significant differences were revealed between groups for FF glucose and E2 levels. CONCLUSION: Present data showed that although FF glucose and E2 levels were constant in PCOS and non PCOS patients, but the FF Hcy levels in PCOS were significantly increased (p=0.01).

The Effects of Chlorpromazine on Reproductive System and Function in Female Rats.

BACKGROUND: Chlorpromazine (CPZ), an antipsychotic drug, is associated with increased risk of sexual dysfunction through increasing prolactin levels. The current study evaluates the effect of CPZ-induced hyperprolactinemia on ovarian follicular growth, gonadotropins, and alteration of ovarian source hormones. MATERIALS AND METHODS: In this experimental study, animals were divided into four groups, control and CPZ (n=8 per group). In the treated groups, CPZ was administered by gavage at doses of 3, 10 and 30 mg/kg per day for 28 days. On day 29 the animals were killed after which histopathological and histomorphometric analyses of the ovaries were performed. We evaluated the levels of prolactin serum, luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2) and progesterone. RESULTS: The ovaries of the test groups showed numerous atretic follicles of various sizes. CPZ caused a significant difference between the test groups and the control group (P<0.05) on the amount of atresia and the size of the normal corpora lutea (CL). The increased dysfunction of the ovaries from the different groups depended on the amount of CPZ administered. The serum concentrations of prolactin and progesterone significantly increased (P<0.05), while the serum concentrations of estradiol, LH and FSH notably decreased (P<0.05), depending on the CPZ dose. CPZ-induced animals had unsuccessful mating and decreased pregnancy rate. CONCLUSION: The present findings suggest that CPZ-induced disturbances not only depend on prolactin level but the increased prolactin level is largely dose-dependent.

Cytotoxic effect of nanosilver particles on testicular tissue: Evidence for biochemical stress and Hsp70-2 protein expression.

Lastly, there are growing evidences that nanosilver (NS) particles highly induce cytotoxic impacts in vitro and in vivo. Here, we analyzed the dose dependent effect of NS on histological changes, biochemical alterations and endocrine statuses, sperm parameters as well as chaperone Hsp70-2 expression. NS particles (50-60nm) were administrated in 3 doses of 0.5, 1 and 5mg/kg, intraperitoneally, for 35 days. The 0.3mL normal saline was administrated in control-sham group. Histological alterations, sperm parameters, serum levels of LH, FSH and testosterone were evaluated. Germinal and Leydig cells RNA damage, Leydig cells steroidogenic foci, the testicular and sperm total antioxidant capacity (TAC), malondialdehyde (MDA), nitric oxide (NO) levels, immunohistochemical (IHC) expression and mRNA level of Hsp70-2 were analyzed. The NS, dose dependently, resulted in enhanced germinal cells degeneration, necrosis, seminiferous tubules atrophy and decreased serum levels of LH, FSH and testosterone. Elevated germinal and Leydig cells RNA damage associated with increased sperm abnormalities were observed in NS-treated groups. Expression of Hsp70-2 was up-regulated in 0.5mg/kg, while its expression was decreased in 1 and 5mg/kg NS-treated groups. Testicular and sperm TAC levels reduced. However, the MDA and NO levels significantly (P<0.05) increased in all NS-treated groups. No histological and biochemical changes were detected in control-sham group. In conclusion, the NS particles exert their pathological impact via affecting testicular antioxidant and endocrine statuses, which in turn lead to diminished expression of Hsp70-2. Ultimately, by this mechanism NS particles adversely impact the cellular RNA, DNA and protein contents.

Chlorpromazine-Induced Hyperprolactinemia on Rat's Uterus.

BACKGROUND: Hyperprolactinemia is a common side effect of antipsychotic drugs that requires further investigation. The current study was designed to evaluate dose-dependent effect of chlorpromazine (CPZ) on hormonal changes and uterine horn histological structure in rats. Moreover, the mammary glands were analyzed to show hyperprolactinemia-induced histological changes. METHODS: Albino Wistar rats (n = 32) were divided into four groups. The first group was set as a control. In the three drug-treated groups (eight rats in each group), CPZ was administered by a gavage at doses of 3, 10, and 30 mg/kg/day for 28 days. One day after the last administration of the drug, the animals were sacrificed. Histopathological and histomorphometrical analyses of the uterine horns and mammary glands were carried out to evaluate dose-dependent effect of CPZ on histological structure. Serum levels of prolactin (PRL), estradiol, progesterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were also evaluated. RESULTS: Remarkable (P < 0.05) elevation was observed in CPZ-administrated animals' uterine horn endometrium, myometrium, and perimetrium thicknesses, and the mammary glands were observed with galactorrhea features. The serum level of progesterone and PRL significantly (P < 0.05) increased, while the serum concentration of LH, FSH, and estradiol was notably (P < 0.05) decreased depending on administrated CPZ dose. No histological and biological changes were occurred in the control animals. CONCLUSION: The present findings suggest that CPZ-induced disturbances not only depend on PRL level and increased PRL level largely depends on administrated doses of the CPZ.

Comparing protective effect of grape seed extract versus atorvastatin on endometriosis in rat model: Evidence for immunohistochemical and biochemical alterations.

Thirty six Wistar albino rats with implant induced endometriosis were randomly divided into six groups of six animals each. The rats in the first group received nothing and were euthanized at day 21. In the second group, rats received nothing and were euthanized at day 36. The third group received atorvastatin (ATV; 5 mg kg(-1) per day, orally) until 21 days from induction of endometriosis, and the fourth group received ATV from the 15(th) day after induction of endometriosis for 21 days. The fifth group received grape seed extract (GET; 450 mg kg(-1) per day, orally) until 21 days from induction of endometriosis. In the sixth group, GET was administered from the 15(th) day after induction of endometriosis for 21 days. The estrogen receptor positive cells (ER+) distribution and angiogenesis were assessed using immunohistochemical and immunoflourescent analyzes, respectively. The active cells with intracytoplasmic carbohydrate content were analyzed. Erα mRNA expression was assessed using semiquantitative real time-PCR and the tissue levels of malondialdehyde (MDA), glutathione peroxidase (GSH-px) and superoxide dismutase (SOD) were evaluated. The GET and ATV-treated animals showed significant reduction in endometriosis-increased ER+ cells distribution as well as significant decrease in Erα mRNA levels (p < 0.05(. Our data suggests that GET exerts a potent inhibitory effect on development of endometriotic implants similar to ATV.

Protective effects of cornus mas extract on in vitro fertilization potential in methotrexate treated male mice.

Current study was aimed to evaluating protective effects of cornus mas fruit extract (CMFE) in mice treated with methotrexate (MTX). For this purpose, 48 young mature male mice were divided into 6 groups. Control group received only normal saline (0.1 mL per day, intraperitoneally), and the second group was administered MTX (20 mg kg(-1) per week, intraperitoneally). The third, fourth and fifth groups received MTX daily oral doses of 250, 500 and 1000 mg kg(-1) CMFE as well as MTX. The sixth group was only given CMFE with a dose of 1000 mg kg(-1) per day, orally, for 35 days. Then, the animals were anesthetically euthanized and the sperms were separated from epididymis. DNA damage level, the amount of malondialdehyde (MDA) as well as in vitro fertility was evaluated. The number of sperms with damaged DNA and MDA level in MTX-treated group showed a significant increase compared to control group (p < 0.05). In groups receiving CMFE along with MTX, DNA damage level and MDA amount suggested a decrease in comparison with MTX group (p < 0.05). Also, in vitro fertilization and embryonic development in MTX-treated group was significantly lower than the control group, and the level of embryonic arresting was higher (p < 0.05). In groups which received CMFE along with MTX, in vitro fertility and embryonic development was higher than MTX group (p < 0.05) and the arrested embryos showed a decrease. This study suggested that cornus mas is able to ameliorate the side effects of MTX.

Protective effects of vitamin E and Cornus mas fruit extract on methotrexate-induced cytotoxicity in sperms of adult mice.

This study was aimed to assess the protective effects of Cornus mas fruit extract (CMFE) and vitamin E (Vit E) on sperm quality parameters in the methotrexate (MTX)-treated mice. Forty-eight young adult male mice (8-12 weeks) were randomly divided into six groups including control and test groups. The control group received normal saline orally , and the test groups were treated MTX (20 mg kg(-1), ip, once weekly), MTX + CMFE (250 mg kg(-1)), MTX + CMFE (500 mg kg(-1)), MTX + CMFE (1000 mg kg(-1)), and MTX + Vit E (100 IU kg(-1), po) for 35 consecutive days. On day 35, after euthanasia the epididymal sperms were isolated. Then the total mean sperm count, sperm viability and motility were determined. The total antioxidant capacity (TAOC) of all experimental groups were also evaluated. The MTX-treated animals showed a significant changes in all parameters of sperm quality assessment compared to the control group. Both Vit E and CMFE were able to protect from MTX-induced effects on sperm maturity and DNA damage. Co-administration of MTX and CMFE and/or Vit E resulted in protection from MTX-reduced TAOC. In conclusion, these data suggested that MTX administration could adversely affect the sperm quality. Moreover, the protective effect of Vit E and CMFE on MTX-induced sperm toxicity was also documented.

Role of mitochondria in ciprofloxacin-induced apoptosis in murine sperm cells.

RATIONALE: Ciprofloxacin (CPFX) has been reported to inhibit cell growth and induce apoptosis in certain eukaryotic cells. The role of the mitochondrial pathway in CPFX-induced apoptosis in cultured murine sperm cells was investigated. METHODS AND RESULTS: Sperm cells (5×10(3) cells/well) from 8-week-old NMRI male mice were cultured in 150 µL of HAM's F10 with 25 mmol/L (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ) HEPES and 10% human serum albumin and were incubated with 50, 100, 200, 400, and 800 µg/mL CPFX for 24 and 36 hours. Cell cytotoxicity, mitochondrial membrane potential (ΔΨM), and concentrations of caspase 3 and caspase 9 were assessed in CPFX-treated cultured murine sperm cells by MTT (3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide) , JC-1 (5, 5á, 6, 6á-tetrachloro-1, 1á, 3, 3á-tetraethylbenzimidazol-carbocyanine iodide)aggregation, and caspase 3 and caspase 9 assays, respectively. Increasing doses of CPFX showed significant cytotoxicity (EC50 = 146.73 µg/mL). Significant loss of ΔΨm was observed in sperm cells treated with ≥50 µg/mL CPFX for 36 hours. Significant increases in caspase 9 and caspase 3 concentrations were also found in cells treated with ≥50 µg/mL CPFX for 24 and 36 hours, respectively (P < .001). CONCLUSIONS: Effects of clinically reachable doses of CPFX on cultured murine sperm cells were investigated and revealed that it may cause sperm cell toxicity by induction of apoptosis through the mitochondrial pathway in the clinically reachable concentrations.

Long-term effect of ciprofloxacin on testicular tissue: evidence for biochemical and histochemical changes.

BACKGROUND: This research studied the effect of ciprofloxacin (CPFX) on spermatogenesis. We aimed to estimate the effect of CPFX on serum levels of testosterone, LH and FSH. MATERIALS AND METHODS: In this experimental study, a total of 24 mice were assigned to controlsham and test groups. We subdivided the test group into low (206 mg/kg) and high (412 mg/kg) dose CPFX groups. Control-sham animals received carboxymethyl cellulose (CMC). All animals were treated orally for 45 days. Cytoplasmic carbohydrate, lipid accumulation, cytoplasmic lipase and alkaline phosphatase (ALP) ratios were examined. Serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH ) and testosterone were measured in the control and test groups. RESULTS: The spermatogenesis cell series exhibited low numbers of cells with periodic acid Schiff (PAS)-positive cytoplasm and higher numbers of cells with lipid-positive foci. The tissue to ALP ratio and germinal epithelium (GE) lipase synthesis increased in CPFX-treated animals. In contrast to the CPFX groups, control animals showed normal cytoplasmic carbohydrate, lipid, lipase and ALP ratios in all cellular layers. In the CPFX-treated groups there was a significantly lower serum testosterone level compared with the control group. The serum levels of FSH and LH in high dosetreated animals decreased. CONCLUSION: Our results suggest that following long time CPFX administration major alterations occur in GE intracytoplasmic biochemistry, which may lead to loss of physiological function and ultimately result in fertility problems. CPFX is able to imbalance serum levels of gonadotropins and testosterone levels by affecting Leydig cells.

Effect of purine nucleoside analogue-acyclovir on the sperm parameters and testosterone production in rats.

BACKGROUND: Acyclovir (ACV), a synthetic purine nucleoside analogue derived from guanosine, is known to be toxic to gonads and the aim of this study was to evaluate the effect of ACV on the sperm parameters and testosterone production in rat. MATERIALS AND METHODS: In this experimental study, forty adult male Wistar rats (220 ± 20 g) were randomly divided into five groups (n=8 for each group). One group served as control and one group served as sham control [distilled water was intraperitoneally (i.p.) injected]. ACV was administered intraperitoneally in the drug treatment groups (4, 16 and 48 mg/kg/day) for 15 days. Eighteen days after the last injection, rats were sacrificed by CO2 inhalation. After that, cauda epididymides were removed surgically. At the end, sperm concentrations in the cauda epididymis, sperm motility, morphology, viability, chromatin quality and DNA integrity were analyzed. Serum testosterone concentrations were determined. RESULTS: The results showed that ACV did not affect sperm count, but decreased sperm motility and sperm viability at 16 and 48 mg/kg dose-levels. Sperm abnormalities increased at 48 mg/kg dose-level of ACV. Further, ACV significantly increases DNA damage at 16 and 48 mg/kg dose-levels and chromatin abnormality at all doses. Besides, a significant decrease in serum testosterone concentrations was observed at 16 and 48 mg/ kg doses. CONCLUSION: The present results highly support the idea that ACV induces testicular toxicity by adverse effects on the sperm parameters and serum level of testosterone in male rats.

Toxic effect of acyclovir on testicular tissue in rats.

BACKGROUND: Acyclovir (ACV), a synthetic purine nucleoside analogue, is known to be toxic to gonads. OBJECTIVE: The current study evaluated cytotoxicity of ACV on histopathological changes in testis tissue and serum testosterone and lipid peroxidation concentrations of male rats. MATERIALS AND METHODS: Animals were divided into five groups. One group served as control and one group served as control sham. In the drug treated groups ACV administered for 15 days. 18 days after the last injection, animals were sacrificed. Histopathological and histomorphometrical analysis of the testis was carried out. Serum levels of testosterone and Lipid Peroxidation and potential fertility of animals was evaluated. RESULTS: Male rats exposed to ACV had significant reduction in serum testosterone concentrations at 16 and 48mg/kg dose-levels (p<0.01). ACV induced histopathological changes in the testis and also increase the mean number of mast cells in peritubular or interstitial tissue in the testis at at 16 and 48mg/kg dose-levels (p<0.01). In addition ACV caused increase of serum level of Lipid Peroxidation at 48mg/kg dose-level (p<0.05). As well ACV decreased potential fertility in male rats. CONCLUSION: The present results highly support the idea that ACV has adverse effect on the reproductive system in male rat.

An investigation on body weights, blood glucose levels and pituitary-gonadal axis hormones in diabetic and metformin-treated diabetic female rats.

Diabetes is a metabolic disorder which affects whole body systems including reproductive system. Diabetes is also a contributing factor to infertility. Metformin is one of the most common drugs to control hyperglycemia. In this study, 36 adult Sprague-Dawley female rats (170-210 g) were divided into 3 groups (control, diabetic and diabetic-treated by metformin). In second and third groups, diabetes was induced by streptozotocin injection (45 mg kg(-1), IP) and the third group was treated by metformin hydrochloride (100 mg kg(-1) day(-1), PO) for 8 weeks. Body weights were compared and blood glucose, gonadotropins and sexual hormones were measured. In diabetic group the blood glucose level significantly (P < 0.05) increased in comparison with that of control and metformin-treated diabetic rats. The results also revealed that, in the untreated diabetic rats, the mean body weights and pituitary-gonadal axis hormones were significantly (P < 0.05) reduced in comparison with the control. Although there were significant (P < 0.05) reduction in mean body weights in metformin-treated diabetic rats, reduction in pituitary-gonadal axis hormones was not as sharp as in untreated diabetic rats and only level of progesterone was significantly (P < 0.05) reduced in comparison with the control. The results of this investigation revealed that there was a clear relationship between experimental diabetes with body weight and pituitary-gonadal axis hormones, and treatment with metformin relatively restored diabetic complications.

The effect of ciprofloxacin on sperm DNA damage, fertility potential and early embryonic development in NMRI mice.

Side effects of ciprofloxacin (CPFX), a widely used broad spectrum antibiotic with fluoroquinolone core, have been reported in different organs. In the present study we sought to elucidate the impact of ciprofloxacin on sperm chromatin integrity and sperm DNA damage using Aniline Blue and Acridine Orange technique, respectively. The fertility potential in male mice was also evaluated. NMRI male mice of 8-week old were included in this study and they were randomly divided into three groups. The first group was received low dose (LD) of ciprofloxacin (206 mg kg(-1), PO) and the second was treated with high dose (HD) of ciprofloxacin (412 mg kg(-1), PO) for 45 consecutive days. The control mice were only treated with oral carboxymethyl cellulose for 45 consecutive days. Sperm cells were removed from cauda epididymis and analyzed for chromatin integrity and DNA damage. In addition, the rate of fertilization, two cell embryos, blastocysts, arrested embryos and their types was examined using zygotes cultured in human tubal fluid - bovine serum albumin (HTF-BSA) medium. Concomitant significant increase in DNA damage and protamine deficiency of the sperm cells in ciprofloxacin treated mice were observed (P < 0.05). In addition, the fertilization rate and embryonic development in treated mice were significantly lower than that of control mice, but the embryo arrest rate in treated mice was significantly higher than that of control group (P < 0.001). In conclusion CPFX was able to induce DNA damage and chromatin abnormalities of sperm cells which could be contributed in the observed low fertilization rate and retarded embryonic development.

The effects of dimethyl sulfoxide and ethylene glycol as vitrification protectants on different cleavage stages of mouse embryo quality.

The effect of modified vitrification was assessed on cellular development capability in mouse embryos cultured in vitro. In this study, 466 embryos (from zygote to morula stages) were vitrified then thawed embryos have been incubated for in vitro farther development up to blastocyst stage. Also, vitrification and thawing procedures were the same for all experimental groups. Mouse different embryonic cleavage stages were vitrified in ethylene glycol (EG) plus dimethyl sulfoxide (DMSO) and sucrose (VS-1) and EG plus DMSO (VS-2) and thawed by directly placing the vitrified drop into sucrose solution (TS) at 37 ˚C. High recovery (72-97%) of morphologically normal embryos was evident following vitrification and thawing. Development of the vitrified morulae into blastocysts (92%) was higher (p < 0.05). The amount of zygote and 2-cell stages that achieved to blastocyst stage was very low. With progressing the embryo cleavage to morula stage, the embryos that reached to blastocyst were increased to its maximum number. We concluded that the modified vitrification procedure supported better survival of morula stage compared to other cleavage stages in mouse embryos.