A caffeine pre-treatment and sole effect of bone-marrow mesenchymal stem cells-derived conditioned media on hyperglycemia-suppressed fertilization.

As a common metabolic disorder, hyperglycemia (HG) affects and disrupts the physiology of various systems in the body. Transplantation of mesenchymal stem cells (MSCs) has been used to control the complications of disease. Most of the therapeutic properties of MSCs are attributed to their secretome. This study aimed to investigate the effects of conditioned media extracted from sole or caffeine pre-treated bone-marrow-derived MSCs on hyperglycemia-induced detrimental impact on some aspects of reproduction. The HG was induced by intraperitoneally injection of streptozotocin (65 mg/kg) and nicotinamide (110 mg/kg). Twenty-four male Wistar rats (190 ± 20 g) were divided into control, HG, and the hyperglycemic groups receiving conditioned media of proliferated MSCs solely (CM) or MSCs pre-treated with caffeine (CCM). During the 49-day treatment, body weight and blood glucose were measured weekly. Finally, HbA1c, spermatogenesis development, sperm count, morphology, viability, motility, chromatin condensation, and DNA integrity were examined. Also, testicular total antioxidant capacity (TAC), malondialdehyde, sperm fertilization potential, and pre-implantation embryo development were evaluated. A one-way ANOVA and Tukey's post-hoc tests were used to analyze the quantitative data. The p < 0.05 was considered statistically significant. The CM and with a higher efficiency, the CCM remarkably (p < 0.05) improved body weight and HG-suppressed spermatogenesis, enhanced sperm parameters, chromatin condensation, DNA integrity, and TAC, reduced HbA1c, sperm abnormalities, and malondialdehyde, and significantly improved pre-implantation embryo development versus HG group. The conditioned media of MSCs solely (CM) and more effectively after pre-treatment of MSCs with caffeine (CCM) could improve spermatogenesis development, sperm quality, pre-implantation embryo development, and testicular global antioxidant potential during hyperglycemia.

Repro-protective role of royal jelly in phenylhydrazine-induced hemolytic anemia in male mice: Histopathological, embryological, and biochemical evidence.

To estimate the repro-protective effect of royal jelly (RJ) on phenylhydrazine (PHZ)-induced anemia's detrimental effects, 24 mature mice were divided into control group (0.10 mL normal saline; intra-peritoneally), RJ group (100 mg/kg/day; orally), experimental anemia (EA) group that received only PHZ (6 mg/100 g/48 h; intra-peritoneally), and RJ + EA (according to the previous prescription) group. After 35 days, testicular histoarchitecture, RNA damage in germinal cells, sperm characteristics, testicular total anti-oxidant capacity and malondialdehyde as well as serum testosterone levels, pre-implantation embryo development and cyclin D1 and c-myc mRNA levels at two-cell, morula and blastocyst stages were analyzed. Spermatogenesis indices were ameliorated following RJ co-administration. Moreover, RJ co-treatment reduced germinal cells RNA damage, improved sperm characteristics, boosted pre-implantation embryo development and restored androgenesis, and oxidant/anti-oxidant status. Co-administration of RJ also decreased mRNA levels of cyclin D1 and up-regulated those of c-myc in two-cell embryos, morulas and blastocysts. The findings suggest that RJ can play a repro-protective role in PHZ-induced anemia in mice through anti-oxidant defense system reinforcement and androgenesis restoration as well as cyclin D1 and c-myc expressions regulation.

Adaptogenic potential of royal jelly in reproductive system of heat stress-exposed male rats.

Testicular heat stress (HS) can lead to testicular tissue destruction and spermatogenesis disturbances. Royal Jelly (RJ) has been introduced as a potent antioxidant. We investigated the effects of RJ on testicular tissue, oxidative stress and sperm apoptosis in HS-exposed rats. Compared to HS-exposed groups, RJ co-treatment could improve testosterone reduction and histopathological damages. The RJ co-administration decreased MDA level in testicular tissue, while TAC and CAT levels were remarkably increased compared to HS-exposed groups. Moreover, significant higher expression level of Bcl-2 and lower expression levels of P53 and Caspase-3 were seen following RJ co-administration compared to HS-exposed groups. Our data suggest that RJ can effectively ameliorate experimental HS-induced testiculopathies in rats through testicular antioxidant defense system restoration and germ cells apoptosis regulation.

Protective effects of licorice extract on ovarian morphology, oocyte maturation, and embryo development in PCOS-induced mice: An experimental study.

BACKGROUND: Polycystic ovary syndrome (PCOS) is an oxidative state resulting in ovarian dysfunction. Licorice is one of the natural antioxidants used for the treatment of infertility. OBJECTIVE: To evaluate the effect of licorice extract on ovarian morphology, oocyte maturation, and embryo development in PCOS-induced mice. MATERIALS AND METHODS: thirty-two female NMIR mice were divided into four groups (n = 8/each): control group receiving no treatment (group I); PCOS group injected with estradiol valerate once daily for 21 days (group II); and experimental groups receiving either 100 mg/kg (group III) or 150 mg/kg (group IV) licorice by gavage along with estradiol valerate once daily for 3 wk. Serum levels of the testosterone and estrogen were measured using ELISA kit. Histological study of ovaries was evaluated, and oocyte maturation, fertilization rate, and embryo development were determined after in vitro maturation. RESULTS: Experimental groups (III, IV) had significantly higher testosterone and estradiol levels compared to the PCOS group (p ≤ 0.001). A significant increase in the number of healthy follicles (primary, preantral follicles) (p = 0.001), corpus luteum (p = 0.001) with significant decrease in the number of atretic follicles (primary, preantral, cystic follicles) (p ≤ 0.001) was seen in the experimental groups. Increase in the fertilization rate (p ≤ 0.001) and blastocyst stage embryos (p = 0.02, p = 0.004) were observed in the experimental groups. CONCLUSION: It appears that the two doses (100 mg and 150 mg) of licorice could decrease ovarian cyst and improve the fertilization rate of oocyte and embryo development in PCOS mice. However, there was no statistically significant difference between the two experimental groups.

Bisphenol-A analogue (bisphenol-S) exposure alters female reproductive tract and apoptosis/oxidative gene expression in blastocyst-derived cells.

OBJECTIVES: One of the major endocrine-disrupting chemicals, bisphenol-S (BPS) has replaced bisphenol-A due to public health anxiety. The present study evaluated low dosage BPS effect on female reproductive potential, hormonal disruption, and gene expression pathways of blastocyst-derived cells. MATERIALS AND METHODS: NMRI female mice (5-6 weeks) in the estrous stage were chosen following vaginal smear examination for estrus cycle detection and BPS (0, 1, 5, 10, 50 and 100 µg/kg) was administrated subcutaneously for twenty-one consecutive days. After the last administration, blood, ovary tissue and oocytes were collected for further examination. RESULTS: BPS induced oxidative stress in ovarian tissue and reduced hormonal status, LH and FSH, even at low concentration. Furthermore, apoptosis was induced in blastocyst derived cells in BPS administrated mice groups even at low BPS concertation, however, P53 and E2f1 expression were downregulated in doses more than 50 µg/kg, which might indicate apoptosis pathway exchange from P53 dependent to p53 independent pathways. IVF outcome was negatively associated with blastocyst apoptosis gene expression, estrogen receptor beta (ERß) as well as oxidative status in ovaries. Finally, Stepwise regression indicated that E2f1, Nrf2, catalase (CAT), and malondialdehyde (MDA) could be chosen as predictor values for hatch percentage in IVF outcome. CONCLUSION: In summary, this study revealed BPS might have detrimental potential in the female reproductive system by oxidation induction and hormonal alteration as well as next generation blastocyst derived cells apoptosis induction. Further studies are recommended for public health assurance of BPS safety especially for female consumed products.

Anatomical and cytohistological study of the pituitary gland in adult turkey.

In order to conduct this study, eight adult turkey heads were obtained. Pituitary glands were harvested following cranial bones removal and examined morphologically and anatomically as well as topographically. Then, tissue sections were prepared and stained using Hematoxylin and Eosin, Alcian blue, orange G and periodic acid-Schiff staining techniques. The results showed that turkey pituitary gland as a pea-sized structure is located in the ventral part of the cerebrum and composed of adenohypophysis and neurohypophysis parts. Moreover, histological analyses revealed that sinusoids are well-developed at the distal part of the adenohypophysis and irregular masses of endocrine cells exist among them. Distributions of basophilic cells in the distal part of adenohypophysis were significantly higher than those of other endocrine cells, while the acidophilic cells had the lowest distribution. Lower and higher numbers of chromophobe cells were also found compared to those of basophilic and acidophilic cells, respectively. These findings were mostly similar to the other birds' pituitary gland anatomical and histological features, but there were also differences in cellular elements distributions along with infundibular cavity topography.

Antioxidant and anti-apoptotic effects of royal jelly against nicotine-induced testicular injury in mice.

This study describes the effects of royal jelly (RJ) on testicular injury induced by nicotine (NIC) in mice. Thirty-six male BALB/c mice were randomly divided into six groups (n = 6). Group 1 received normal saline, group 2 received 100 mg/kgBW/day RJ, groups 3 and 4 received NIC at doses of 0.50 and 1.00 mg/kgBW/day, respectively, and groups 5 and 6 received NIC at doses of 0.50 and 1.00 mg/kg BW/day, respectively, plus RJ. Following 35 days, the serum level of testosterone, histopathological changes, germ cell apoptosis, proliferating cell nuclear antigen (PCNA), malondialdehyde (MDA) content, and antioxidant indexes including total antioxidant capacity (TAC) and catalase (CAT) activity were determined. In addition, the mitochondria-dependent apoptosis was investigated by assessing the Bcl-2, p53, and Caspase-3 mRNA levels expression by reverse transcription-PCR (RT-PCR). Compared to NIC receiving groups, the concomitant administration of RJ could protect the testosterone reduction and histological damages. After RJ treatment, the level of tissue MDA content decreased, while tissue TAC and CAT levels were remarkably increased compared to NIC-exposed groups. Remarkable higher TUNEL-positive germ cells and low PCNA index were observed in NIC receiving groups. Besides, the expression level of Bcl-2 was significantly higher and the p53 and Caspase-3 levels were significantly lower in the RJ co-administration groups than NIC-only receiving groups. Our results confirmed that RJ effectively protects the testis against NIC evoked damages by antioxidant and anti-apoptotic effects involving the up regulation of the antioxidant status, mitochondria-dependent apoptosis pathway prevention, and the proliferating activity improvement.

Ameliorative effect of omega-3 on spermatogenesis, testicular antioxidant status and preimplantation embryo development in streptozotocin-induced diabetes in rats.

PURPOSE: The present study was done to determine the ameliorative effect of polyunsaturated fatty acid omega-3 against experimental diabetes-induced damages on testicular tissue, sperm parameters and preimplantation embryo development in rat model. METHODS: Thirty-two mature male rats were divided into two control and test groups. The experimental diabetes (50 mg kg-1 streptozotocin, ip) was induced in test group and subdivided into non-treated diabetic, 300 and 600 mg kg-1 omega-3-treated (orally by gavage) groups. The rats in control group received 0.5 ml saline using intra-gastric gavage. Following 45 days, general histopathological changes, serum level of testosterone, inhibin B, glucose, and sperm parameters, testicular total antioxidant capacity (TAC) and malondialdehyde (MDA) content were analyzed. The mitochondria-dependent apoptosis was investigated by assessing the Bcl-2 and caspase-3 expression as well as DNA fragmentation. Finally, the in vitro fertilization (IVF) potential was examined by evaluating preimplantation embryo developing. RESULTS: The omega-3 significantly ameliorated the diabetes-induced histological damages, diminished serum level of glucose, testicular MDA content, and enhanced the serum testosterone, inhibin B and testicular TAC. The animals in omega-3-treated groups exhibited a significant (p < 0.05) up-regulation in Bcl-2, as well as remarkable (p < 0.05) down-regulation in caspase-3 expression compared to non-treated diabetic rats. Moreover, the omega-3 maintained DNA integrity, improved sperm quality as well as preimplantation embryo development. CONCLUSION: Our data showed that the omega-3 (especially at 600 mg kg-1 dose level) effectively ameliorates the experimental diabetes-induced infertility in rats by up-regulating the testicular endocrine and antioxidant statuses, preventing mitochondria-dependent apoptosis pathway and potentially improving the sperm quality.

Attenuation of Methotrexate-Induced Embryotoxicity and Oxidative Stress by Ethyl Pyruvate.

BACKGROUND: Methotrexate (MTX), as an anti-folate agent, is widely used in the treatment of rheumatic disorders and malignant tumors, however it damages reproductive sys- tem in mice. The aim of this research was to study the effects of ethyl pyruvate (EP) on embryo development and oxidative stress changes in the testis of mice treated with MTX. MATERIALS AND METHODS: In this experimental study, thirty-two adult male Naval Medical Research Institute mice, with average weight of 26 ± 2 g, were divided into four groups. The first group (control) received distilled water (0.1 ml/mice/day), while the second group was intraperitoneally (IP) treated with 20 mg/kg MTX once per week. The third group was IP treated with 40 mg/kg/day EP, and the fourth group was IP treated with both 20 mg/kg MTX and 40 mg/kg/day EP for 30 days. At the end of treatment fertilization rate and embryonic development were evaluated. Differences between these groups were assessed by ANOVA using the SPSS software package for Windows with a Tukey-Kramer multiple post-hoc comparison test. RESULTS: MTX treatment caused significant (P<0.05) increase in malondialdehyde (MDA) and reduced catalase (CAT), as well as leading to in vitro fertilization (IVF) and embryonic development. The improved effects of EP on the IVF were determined by the reduced level of MDA (index of oxidative stress) and significant increased level of CAT (a key antioxidant). We observed significant increase in fertilization rate and embryonic development in the treated group with both MTX and EP. CONCLUSION: It is suggested that EP can be useful in ameliorating testicular damages and embryotoxicity induced by MTX. These effects could be attributed to its antioxidant properties.

Protection against Cyclosporine-Induced Reprotoxicity by Satureja khuzestanica Essential Oil in Male Rats.

BACKGROUND: The effects of cyclosporine (Cs), a fungal cyclic polypeptide with potent immunosuppressive activity, on fertility have assumed greater significance with the increasing numbers of transplantations being performed all over the world. Current study was undertaken to investigate the potential of Satureja khuzestanica Essential Oil (SEO) as an antioxidant to mitigate Cs-induced reprotoxicity. MATERIALS AND METHODS: In this experimental study (April-July 2012), thirty-two adult male Wistar rats were randomly divided into 4 groups of 8 animals each. Two groups of rats were administered Cs [40 mg/kg/day, per oral (p.o.)] for 45 days. One of these groups received SEO (225 mg/kg/day, p.o.) four hours after Cs administration. A vehicle-treated control group and a SEO control group were also included. Epididymal sperm characteristics, in vitro fertilizing capacity as well as embryo development were evaluated. For statistical analysis, one-way ANOVA and Tukey's post-hoc test were used, and the value of P<0.05 was considered as the criterion for statistical significance. RESULTS: Sperm count and viability along with fertilization and blastocyst development rates were significantly decreased by Cs treatment. Moreover, Cs-treated group showed significant increases in DNA damage, protamine deficiency of the sperm cells and proportion of spermatozoa with cytoplasmic droplet. Notably, aforementioned parameters were improved to near normal level by SEO co-administration. CONCLUSION: These results suggest that SEO has a protective action against Cs-induced reprotoxicity in a rat model.

Protective effect of royal jelly on fertility and biochemical parameters in bleomycin- induced male rats.

BACKGROUND: Bleomycin (BL) is a glycopeptide antibiotic obtained from the bacterium Streptomyces verticillus which is routinely used for treatment of human cancers. Royal jelly (RJ) is a production from the hypo pharyngeal, mandibular and post cerebral glands of nurse bees. RJ consists of 66% water, 15% sugars, 5% lipids, and 13% proteins, essential amino acids and vitamins. OBJECTIVE: The aim of present study was to evaluate protective effect of royal jelly on sperm parameters and malondialdehyde (MDA) production in rat. MATERIALS AND METHODS: Forty adult male wistar rats (220±20gr) were randomly divided into 4 groups (n=10). Control group (CG) received normal saline 10 ml/kg twice a week with Intraperitoneal (I.P) for 48 days (0.3 ml/rat(. Royal Jelly group (RJG) received jelly (100 mg/kg daily) for 48 days orally. Bleomycin group (BLG) received BL (10 mg/kg twice a week) with I.P for 48 days. Royal Jelly+ Bleomycin group (RJ+BLG) received royal Jelly (100 mg/kg /day) orally concomitant with BL administration. Sperm count, motility, and viability were investigated and chromatin quality and DNA integrity were also analyzed. Serum testosterone and MDA concentrations were measured as well. RESULTS: BL caused decline significantly (p<0.05) sperm count, sperm viability, motility as well as testosterone concentration compared to control group while significant (p<0.05) increases in immature sperm, sperm with damaged DNA and MDA concentration were announced in BL in comparison with CG and RJ+BLG. Royal jelly improved Bleomycin-induced toxicity on sperm parameters and testosterone and MDA concentrations. CONCLUSION: The present results support the idea that BL adversely affects sperm parameters and MDA and the RJ with antioxidant properties has positive effects on these parameters. This article extracted from M.Sc. thesis. (Tayebeh amirshahi).

Protective effect of ethyl pyruvate on epididymal sperm characteristics, oxidative stress and testosterone level in methotrexate treated mice.

BACKGROUND: Methotrexate (MTX) is an anti-metabolite drug widely used in treatment of neoplastic disorders, rheumatoid arthritis and psoriasis. The ester derivative, ethyl pyruvate (EP) is stable in solution and should function as an antioxidant and energy precursor. This study was conducted to evaluate the protective role of EP on sperm parameters, testosterone level and malondialdehyde (MDA) production in mice treated with MTX. METHODS: 32 adult male NMRI mice weighing 26±2 g were divided into 4 groups. Group 1 received 0.1 ml/mice/day of distilled water intraperitoneally for 30 days (ip). Group 2 was treated with methotrexate at a dose of 20 mg/kg once a week (ip) for 30 days. Group 3 was treated with ethyl pyruvate at a dose of 40 mg/kg/daily (ip) for 30 days. Group 4 was treated with methotrexate (20 mg/kg) once a week simultaneously with ethyl pyruvate 40 mg/kg for 30 days. The results were analyzed by oneway ANOVA. A p<0.05 was considered to be significant. RESULTS: The results showed significant (p<0.05) decrease in sperm count and sperm motility as well as testosterone concentration while sperm with damaged DNA and MDA concentration in mice treated with MTX in comparison with control and MX+EP groups increased significantly (p<0.05). Instead, MTX+EP group caused partial amelioration in all parameters mentioned above. CONCLUSION: Based on the present study, it can be concluded that MTX induced toxicity in sperm parameters and serum level of testosterone and increased MDA level. EP with its antioxidant properties could be administrated during treatment with MTX due to its protective effects on sperm parameters, plasma testosterone levels and lipid peroxidation.