Hydrostatic pressure improves in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries

Background: Cryopreservation has limited successes and in-vitro maturation is used to improve its results. Hydrostatic pressure (HP) plays an important role in follicular development. Objective: This study was designed to examine the effects of HP on in-vitro maturation of oocytes and cell death in cumulus cells derived from vitrified-warmed mouse ovaries. Materials and Methods: Preovulatory follicles were harvested from non-vitrified and vitrified-warmed 6-8 week-old female NMRI mouse ovaries and randomly assigned to following groups: non-vitrified (control), non-vitrified with HP exposure (treatment I), vitrified-warmed (treatment II) and vitrified-warmed with HP exposure (treatment III). The follicles of treatments I and III were subjected to HP (20 mmHg) for 30 min and after that all groups were cultured for 24h and assessed for in-vitro maturation of oocytes. The viability and apoptosis of cumulus cells and oocytes were assessed using supravital nuclear staining and TUNEL assay, respectively. Results: Oocytes harvested follicles in both control and treatment II had a significantly lower percentage of metaphase II oocytes (MII) than the treatment I and III (23.5±3.1, 15.03±4.6 and 32.7±3.2, 25.5±4.6; respectively) (p<0.05). Viability of the cumulus cells reduced in treatment I, II and III (83.4, 83.3 and 77.7%) compared to control (86.9%), (p<0.05). The apoptotic index in cumulus and oocyte complexes in treatments I and III (10.7±0.8 and 15.3±0.8) was higher than in control and treatment II (6.7±0.5 and 9.7±0.5) (p<0.05). Conclusion: These results demonstrate that HP had a mild effect on cell death incidence in cumulus cells without any effect on oocyte. However, it can be used as a mechanical force to improve in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries

Possible involvement of calcium channels and plasma membrane receptors on Staurosporine-induced neurite outgrowth

Staurosporine as a protein kinases inhibitor induced cell death or neurite outgrowth in PC12 cells. We investigated the involvement of calcium channel and plasma membrane receptors on staurosporine inducing neurite outgrowth in PC12 cells. PC12 cells were preincubated with NMDA receptor inhibitors (1.8 mM ketamine and 1μM MK801, treatment 1) or L-Type Calcium channels (100 μM nifedipine and 100 μM flavoxate hydrochloride, treatment 2) or calcium-calmoduline kinasses (10 μM trifluoprazine, treatment 3) and nifedipine, MK801, flavoxate hydrochloride and ketamine (treatment 4 or without pretreatments (control). Then, the cells were cultured in RPMI culture medium containing 214nM staurosporine for induction of neurite outgrowth. The percentage of Cell cytotoxicity and apoptotic index was assessed. Total neurite length (TNL) and fraction of cell differentiation were assessed. After 24h, the percentage of cell cytotoxicity were increased in treatments 1, 2 and 4 compared with control (p<0.05). After 6h, apoptotic index was similar between all treatments. After 12h, apoptotic index were increased in treatment 4 compared with control (p<0.05). After 24h, apoptotic index were increased in treatments 1, 2 and 4 compared with control (p<0.05). TNL were decreased in treatments 1, 2 and 4 compared with control in different times of assessment (6, 12 and 24 h) (p<0.05). The fraction of cell differentiation were decreased in treatments 1, 2 and 4 compared with control (p<0.05). It can be concluded that the possible involvement of L-type calcium channel and the N-methyl D-aspartate receptor on staurosporine-induced neurite outgrowth process in PC12 cells

Protective Effect of Royal Jelly against Renal Damage in Streptozotocin Induced Diabetic Rats

Background: Royal jelly has been shown to have antioxidant and antidiabetic effects. The objective of this study was to evaluate the protective effect of RJ against kidney damage in streptozotocin induced diabetic rats. Methods: Thirty two male Wistar rats were divided randomly into four groups (n=8 per group). Normal control and diabetic control groups received 1cc/day distilled water, normal RJ-treated and diabetic RJ-treated groups received 100mg RJ/kg body weight daily. Diabetes was induced by intraperitoneal injection of streptozotocin. At the end of the experiment, urine and kidney samples were collected for biochemical and histopathological analysis. Results: The results showed that diabetes could increase levels of urine urea, total protein and albumin significantly, and could decrease the levels of creatinine and uric acid in urine. In the kidney tissue homogenates, catalase activity and antioxidant power were significantly lower, whereas malondialdehyde levels were significantly higher in diabetic group when compared with control group. Diabetic rats showed severe histological changes in kidney tissues. Treatment of diabetic rats with RJ improved significantly all of these parameters. Conclusion: The present study revealed that treatment with RJ resulted in significant improvement in histopathological alterations in kidney tissue and urine parameters of diabetic rats. This could be due to its antioxidant activity and the ability of RJ for scavenging the free radicals released in diabetes. These findings suggest that RJ has protective effects on kidneys affected by diabetes mellitus

Polycystic Ovary Induction in Mouse by Testosterone Enanthate

Background &Objective: Polycystic ovary is the most common cause of infertility in Women. Animal models are required for understanding the pathogenesis of polycystic ovary. The objective of this study then was to develop an animal model for inducing the polycystic ovaries using testosterone enanthate.Materials & Methods: In this study, for inducing the polycystic ovary phenotype, female rats about12-14 days-old were injected daily with testosterone enanthate for 2 and 4 weeks (experiment groups: 1 and 2), while the control groups (1 and 2) were injected only with vehicle.The ovaries from both groups were fixed and then were used for histological studies.Results: Testosterone enanthate treatment causes the histological changes in mouse ovary and significantly increased the percentage of preantral and cystic follicles and decreased the percentage of antral follicles in the experiment group, comparing with the control group (P<0.05).Conclusion: It concluded that testosterone enanthate can induces polycystic ovary in mouse

The Effect of Repaglinide on Polycystic Ovary Induced by Testosterone Enanthate in NMRI Mice

Background & Objective: Poly Cystic Ovarian Syndrome (PCOS) is the most common cause of infertility due to anovulation in women. One of the treatments of this syndrome is using Insulin sensitive drugs. The main objective of this project is to analyze the effect of Repaglinide on the histomorphology of polycystic ovary. &nbsp;Materials & Methods: In this study, PCOS is induced by the injection of Testosterone Enanthate in immature mice aged five to seven weeks old for four weeks. Then, the mice were divided in to two groups of control and case. The control group had no injection. In the case group, each mouse received 30 &micro;gr Repaglinide. Body weight, the ratio of ovary weight to body weight, and ovarian diameter were measured and the changes in histomorphology of the ovary, characteristics of ovarian tissue, and the number of follicles were investigated. T- Test was used in this study (T-test, P&pound;0.05. ( Results: The result revealed that Repaglinide had a reducing effect on body weight, the ratio of ovary weight to body weight, and the diameter of ovary (P&pound;0.05). Repaglinide also caused a decrease in Tunica albuginea changes, hyperthecosis, the number of degenerated oocyte, granolusa cell, picnosis and hyper vascularization. The average of primordial, primary, pre-antral, cystic, and atretic follicles are significantly decreased in the case group compared to the control group (P&pound;0.05). Conclusion: Repaglinide can compensate the PCOS ovarian damages. Further studies are recommended in this field

A Histomorphological Comparative Effects of Metformin, Pioglitazone, Repaglinide and Acarbose on Polycystic Ovary in Mice

Background & Objective: Polycystic ovary syndrome (PCO) is one of the most common causes of infertility due to anovulatory in women. The use of insulin-sensitizing agents was one of the treatments for this syndrome. The main purpose of this study was to evaluate the effects of Metformin, Pioglitazone, Ripaglinide and Acarbose on histomorphological changes of polycystic ovaries. Materials & Methods: In this study, Polycystic ovaries were induced by injection of testosterone enanthate (TE) in immature female mice, for four weeks, then they had been divided into five groups; the control, Metformin, Pioglitazone, Repaglinide and Acarbose groups. Body weight, the ratio of ovary-to-animal weight, ovarian diameter, histomorphological changes of ovaries and characteristics of ovarian tissues were studied. Tukey test was used to calculate these parameters (P≤0.05). Results:TE significantly increased the percentage of cystic Follicular and decreased follicular growth compared to treatment groups. The body weight, the ratio of ovary-to-animal weight and ovarian diameter, in all groups showed a significant decrease compared to the control group (p<0.05). In Metformin and Pioglitazone treated group, the number of degenerated oocytes, pyknotic-granlosa cells and vascularization were decreased and luteinization can be seen only in these groups. There is a significant reduction in mean growth of primordial, primary, pre-antral, cystic and atretic follicles in the treatment group. However, the mean number of pre-antral follicle showed a remarkable increase. The average number of antral follicles increased in metformin and pioglitazone groups (p<0.05). The results showed that Metformin and Pioglitazone groups cause a meaningful decrease in the ratio of ovary-to-animal weight (p<0.05). Conclusion: According to these results, Metformin and Pioglitazone have the same effects and can compensate for the damages due to PCOs. These drugs can develop follicular growth. Repaglinide can compensate for the damages in some cases, and, Acarbose has a negative effect on follicular growth

The Study of Retinoic Acid Effects on Testicular Development of NMRI Mice

Background & Objectives: Retinoids are important molecules that regulate crucial processes of development in all vertebrates. In this article, we study the effect of retinoic acid on testis development. Materials & Methods: A group of newborn NMRI mice was chosen to receive intraperitoneally injections of 25 mg/Kg.b.w of retinoic acid and the control group had no injection. After 21 days, the male animals were isolated and sacrificed in 60 days postpartum, and testes were removed from their bodies. Apparent characteristics of the testes of seven mice from each group were observed. In the light microscopic study, these epithelial cells of the tubules were counted and the diameter of seminiferous tubules were measured with statistical T-test analysis and were compared with control group. Results: The findings showed that the weight of the testes in animals that were affected by retinoic acid did not change significantly compared to control group. In germ epithelium of testis, the thickness of the epithelium and the number of the spermatogonia, round and elongated spermatids has decreased significantly compared to the control group (P<0.05). Conclusion: According to the findings of this article, using the retinoic acid after birth has influence on testis development and its seminiferous tubules epithelium. Retinoic acid has an adverse effect on the cell divisions in seminiferous epithelium, therefore the number of affected germ cells decrease. Therefore, the use of vitamin A and its synthetic derivatives like retinoic acid for pregnant women should be done with caution

Ovary stereological features and serum biochemical factors following induction of polycystic ovary syndrome with testosterone enanthate in mice: An experimental study

Background: Polycystic ovary syndrome (PCOS) is an endocrine disorder featured by insulin resistance and hyperandrogenism. Testosterone enanthate can induce PCOS in mice models. Objective: We investigated the ovary stereological features along with the oxidative stress and inflammatory factors in mice following PCOS induction using testosterone enanthate. Materials and Methods: Twelve female NMRI mice (3 wk old) were divided into 2 groups (n=6/each): Control and PCOS. PCOS was induced through daily injections of testosterone enanthate (1 mg/100g subcutaneous s.c for 5 wk). Finally, ovaries were studied stereologically. The serum levels of the follicle-stimulating hormone, luteinizing hormone, testosterone, interleukin-6, and tumor necrosis factor-&alpha; were measured using ELISA kit. Serum levels of Malondialdehyde and the antioxidant capacity were measured relatively using thiobarbituric acid and ferric reducing antioxidant power assay. Results: The mean total volume of ovary and the mean volume of cortex (p<0.001), volume of oocyte in the preantral (p=0.011) and antral follicle (p=0.015), thickness of zona pellucida (p=0.016), the number of antral follicles (p=0.012), the serum levels of follicle-stimulating hormone (p<0.001) and the antioxidant capacity (p=0.020) reduced significantly in the PCOS group compared to the control. The number of primary (p=0.017) and preantral (p=0.006) follicles and the serum levels of testosterone (p<0.001), Luteinizing hormone (p=0.002), Malondialdehyde, Interleukin 6 and Tumor necrosis factor-&alpha; (p<0.001) showed a significant increase in the PCOS group compared to the control. Conclusion: Testosterone enanthate induced PCOS causes stereological features in the ovary, increases the oxidative stress and inflammatory markers in mice