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Showing 54 results for Apoptosis

Fahime Sadat Kamali, Rasoul Shahrooz, Golamreza Najafi, Mazdak Razi,
Volume 0, Issue 12 (12-2019)

Background: Paraquat (PQ), as a pyridine compound, is widely used worldwide to control annual weeds. The oxidative stress caused by PQ can cause deleterious changes in the testicular tissue.
Objective: An investigation on the protective effects of Crocin (CCN) against PQ-induced oxidative damages and apoptotic indices in testicular tissue. 
Materials and Methods: Twenty-eight adult male albino mice (20-25 gr) were divided into four groups (n = 7/each). The control group received 0.1 ml/day of normal saline by intraperitoneal injection (IP); sham-control group received PQ 5 mg/kg/day, IP, and the experimental groups received PQ (CCN+PQ) and CCN-sole (200 mg/kg/day, IP), respectively, for 35 continuous days. At the end of the treatment period, the testes were dissected out and used for biochemical, molecular, and histological analyses. The expressions of tumor suppressor p53, B-cell lymphoma 2 (bcl-2), and caspase-3 were considered as hallmark factors of mitochondria-dependent apoptosis. Moreover, the testicular superoxide dismutase (SOD) and malondialdehyde (MDA) were evaluated as key biomarkers for oxidative stress.
Results: The PQ significantly (p< 0.02, p< 0.01) diminished the spermatogenesis indices and SOD, increased MDA levels, and enhanced the apoptosis-related gene expression. However, the co-administration of CCN and PQ significantly (p< 0.01, p< 0.01, p< 0.02) ameliorated the spermatogenesis ratio, upregulated the SOD level as well as bcl-2 expression, and reduced the MDA content and apoptosis vs the PQ-sole group.
Conclusion: This study showed that the antioxidant properties of CCN enable to ameliorate the PQ-induced destructive effects by upregulating the testicular structure, antioxidant and apoptotic status.
Ashok Agarwal, Sushil Anandh Prabakaran,
Volume 3, Issue 1 (7-2005)

Infertility is one of the most stressful conditions amongst married couples. Male factor infertility is implicated in almost half of these cases. Recent advances in the field of reproductive medicine have focused the attention of many researchers to consider reactive oxygen species (ROS) as one of the mediators of infertility causing sperm dysfunction. Although, ROS is involved in many physiological functions of human spermatozoa, their excess production results in oxidative stress. Mitochondria and sperm plasma membranes are the two locations of ROS production that involves complex enzyme systems such as creatine kinase and diaphorase. ROS causes damage to the spermatozoa DNA, resulting in increased apoptosis of these cells. The production of ROS is greatly enhanced under the influence of various environmental and life style factors such as pollution and smoking. An effective scavenging system is essential to counteract the effects of ROS. Various endogenous antioxidants belonging to both enzymatic and non-enzymatic groups can remove the excess ROS and prevent oxidative stress. Since, ROS is essential for the normal sperm physiology, rationale use of antioxidants is advocated.
Tahmineh Peirouvi, Gholamhossein Farjah, Jafar Soleimani Rad, Marefat Ghaffari Novin,
Volume 5, Issue 4 (7-2007)

Background: Phospholipids are distributed asymmetrically between inner and outer leaflets of the plasma membrane of live cells. Early during apoptosis, this asymmetry is disrupted and phosphatidylserine becomes exposed on the outside surface of the plasma membrane. There is little information about the effects of vitrification on apoptosis.
Objective: The aim of the present study was to evaluate the effect of vitrification on apoptosis of subfertile and fertile men.
Materials and Methods: In this study, semen samples were collected from subfertile (n=20) and fertile men (n=10) after 48h abstinence of intercourse. After semen analysis according to WHO criterias, each semen sample was divided into two portions. First portion was assessed by Annexin V-flous staining kit for showing apoptosis in subfertile and fertile men and second portion was assessed after vitrification-thawing. Results were analyzed by Paired t-test and Independent t-test.
Results: After vitrification-thawing, mean percentage of apoptotic spermatozoa has increased 6 and 3 times in subfertile and fertile men respectively. This difference is significant.
Conclusion: Vitrification-thawing could disrupted membrane asymmetry and caused apoptosis. Therefore, it will cause reduction of functional spermatozoa in access of Assisted Reproduction Technologies (ART).
Arash Khaki, Mahnaz Heidari, Marefat Ghaffari Novin, Amir Afshin Khaki,
Volume 6, Issue 3 (7-2008)

Background: Ciprofloxacin is a commonly prescribed antibiotic in the treatment of genitourinary tract infection.
Objective: The aim of this study was to investigate the effects of ciprofloxacin on testis apoptosis and sperm parameters in rat.
Materials and Methods: Twenty male Wistar rats were selected and randomly divided into two groups; control (n=10) and experimental (n=10). The experimental group was orally received 12.5 mg/kg ciprofloxacin daily for 60 days and the control group just received water and food. Rats were then killed and sperm removed from cauda epididymis and analyzed for sperm motility, morphology, and viability. Testis tissues were also removed and prepared for TUNEL assay to detect apoptosis.
Results: Results showed that ciprofloxacin significantly decreased the sperm concentration, motility (p<0.05) and viability (p<0.001). In addition, ciprofloxacin treatment resulted in a significant decrease in the number of spermatogenic cells (spermatogonia, spermatocyte, spermatid and sperm) in the seminiferous tubules when compared with the control group. The apoptotic germ cells per seminiferous tubular cross section was significantly increased in the experimental group (15.11±3.523) as compared with the control group (7.3±0.762) (p<0.05).
Conclusion: It is concluded that ciprofloxacin has the toxicological effects on reproductive system in male rats.
Forouzan Absalan, Mansoureh Movahedin, Seyed Javad Mowla,
Volume 6, Issue 4 (7-2008)

Background: In most mammals, the testis is always maintained at a lower temperature than that in the abdomen, and exposure of the testis to body temperature causes degeneration of germ cells.
Objective: In this research, the long effect of heat exposure on sperm parameters and microstructure of mouse testis were investigated. Cryptorchid mouse were induced by exposure to abdominal heat.
Materials and Methods: Immature mice were anesthetized and a small incision was made in the abdominal skin, then fat pad at the upper end of testis was sutured to peritoneum. Weight of testis, spermatogenic cell numbers, tubular ectasis (rate of tubular lumen comparing to the thickness of germinal epithelium) as well as epididymal sperm parameters were measured.
Results: The results showed that spermatogenesis was arrested and testicular weights, seminiferouse tubular diameters and epididymal sperm parameters were significantly reduced in the bilateral undescended testis compared with unilateral undescended testis and the control mice. However, complete depletion of seminiferous tubules and absence of germ cells was not found in the animals.
Conclusion: In general, high temperature caused a decreased in all analyzed parameters except spermatogonial cell number probably due to the apoptosis and these changes significantly increase in bilateral groups compared with unilateral groups. We believe that the present model is a suitable tool for enrichment of spermatogonial stem cells, also it is useful for treatment of cryptorchidism and further biological research on spermatogenesis.
Hassan Hassani Bafrani, Hidekazu Saito,
Volume 6, Issue 5 (7-2008)

Background: The mitogen-activated protein kinase (MAPK) pathway is one of the major signaling pathways that transmit intracellular signals initiated by extracellular stimuli to the nucleus. The stress-activated protein kinase (SAPK)/c-Jun NH2-terminal kinase is a subfamily of MAP kinases implicated in cytokine and stress responses.
Objective: In this study, we have examined total and phosphorylated c-Jun in the mural and cumulus granulosa cells, and investigated also whether c-Jun can be responsible for the difference in the expression of apoptosis between mural and cumulus regions.
Materials and Methods: A total of 14 consecutive couples participating in IVF program were investigated. Aspirated follicular fluid was transferred into tissue culture dishes and oocyte-cumulus cells complexes were isolated. The cells were centrifuge and fixed with Bouin’s solution and then were put on a glass slide. After fixation, the slides were stained by immunocytochemistry method. The incidence of apoptotic granulosa cells was examined by a fluorescence microscope.
Results: The incidence of apoptotic granulosa cells was 1.27 ± 0.12 in the mural region and 0.38 ± 0.07 in the cumulus regions. All mural and cumulus cells expressed total c-Jun in 7 patients while phosphorylated c-Jun was also expressed in all cells of the other 7 patients. There was no difference between apoptotic and nonapoptotic cells in the expression of total and phosphorylated c-Jun.
Conclusion: C-Jun may not be responsible for apoptotic effect on mural and cumulus cells.
Tahereh Talaei-Khozani, Zahra Borzoei, Soghra Bahmanpour, Jaleh Zolghadr, Sedigheh Dehbashi, Hamid Reza Zareh,
Volume 9, Issue 2 (7-2011)

Background: Recurrent spontaneous abortion impacts almost 1% of couples. The sera from women with unexplained recurrent spontaneous abortion (URSA) have toxic effects on embryos that grow in the uterus. Therefore, the abnormal condition of the uterus may also affect sperm qualities.
Objective: The objectives of this study were to search if these sera could induce DNA denaturation in sperm nuclei and also it could reduce sperm motility.
Materials and Methods: Sera of 20 women with URSA history and sera from 20 women with at least two healthy children were added to the sperms samples from 20 healthy men for 2 hours. The sperm motility was assessed after incubation with sera. The samples were stained with Tdt mediated dUTP nick end labeling (TUNEL) assay for DNA fragmentation. The samples were analyzed with flow cytometry and the percentage of the TUNEL positive sperms were calculated. The data were analyzed by t-test.
Results: The incubation of the sperm samples in sera with URSA lead to a decrease in the percentage of the motile sperm from 55% in control to 41% in the treated group, significantly (p=0.038). The percentage of the sperm with abnormal fragmented DNA increased after incubation with URSA (26.6%) compare to the control (21.2%); however, it was not significant.
Conclusion: It seems that sera from URSA patients could not induce a significant increase in the percentage of the sperms with nuclei contain DNA fragmentation. However, the sera of women with URSA could affect the fertility rate by reduction of the sperm motility
Khadijeh Foghi, Marefat Ghaffari Novin, Zahra Madjd Jabbari, Tohid Najafi, Mohammad Hasan Heidari, Abouzar Rostampour Yasoori,
Volume 9, Issue 4 (7-2011)

Background: Non obstructive azoospermia (NOA) is one of the causes of male infertility in which spermatogenesis process is disturbed. Recent studies suggested the possible role of endothelial nitric oxide synthase (eNOS) in spermatogenesis process.
Objective: The aim of the present study is to evaluate the expression of eNOS in human testicular tissue in men with NOA and men with normal spermatogenesis by using immunocytochemistry.
Materials and Methods: In this case-control study, testicular biopsies were obtained from 10 men with NOA and 7 men with normospermia who were attended to infertility center for diagnosis or infertility treatment. Immunohistochemistry was used to localize the isoform of eNOS in these tissues and the intensity of staining was semi quantitively assessed. In addition, the histopathological evaluation was examined in both groups.
Results: The isoform of eNOS enzyme activity was detected in the cytoplasm of sertoli and leydig cells in both groups. There was, however, a considerable variability in the intensity of staining between two groups. The expression of eNOS in Leydig cells in control group was significantly (p<0.05) higher than those in the NOA group. In contrast, expression of eNOS in Sertoli cells in NOA was more than those in the control group. eNO Simmune staining was absent in the normal germ cells but was intense in the abnormal germ cells with piknotic neucleous. The most histopathological finding were hypospermatogenesis (27.2%), Sertoli cell only syndrome (18.1%) and tubular fibrotic (13.6%).
Conclusion: These results suggested that increase level of eNOS may play an important role in the apoptosis process in the abnormal germ cells and disturbance of spermatogenesis process.
Zahra Rashidi, Mehri Azadbakht, Mozafar Khazaei,
Volume 10, Issue 3 (7-2012)

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.

Tahereh Mirjalili, Seyed Mehdi Kalantar, Maryam Shams Lahijani, Mohamad Hasan Sheikhha, Alireza Talebi,
Volume 11, Issue 1 (4-2013)

Background: Methamphetamine (MA) is a potent psychomotor stimulant with high abuse and addictive potential. MA is a neurotoxic drug which is widely abused by females of childbearing age, raising serious public health concerns in terms of exposure of the fetus to the drug. Neurotoxic effects of MA on adult are well known, such as dopaminergic nerve terminal degeneration and cell death in regions of brain in some doses.
Objective: In the present study, we examined effect of prenatal MA exposure on mouse fetuses.
Materials and Methods: In this study, forty 8-12 week-old NMRI female mice were used which were mated with male mice in serial days. When sperm plug was observed it was designated as gestational day (GD) 0. Pregnant mice were individually housed in plastic cages. Pregnant mice were divided into four groups: in first group 10 mg/kg /day MA, in second group 5 mg/kg /day MA and in third group saline were injected subcutaneously from GD 6 to GD 14, corresponding to organogenesis period, while fourth or control group were without injection. On GD 14 fetuses were removed and accomplished chromosome preparation from fetal liver. Then fetal were fixed in formalin for brain hematoxilin and eosine staining and TUNEL assay.
Results: We observed morphological abnormality including exencephal fetus in 5mg/kg MA group and premature fetuses in 10 mg/kg MA group. Also brain histological study showed subarachnoid hemorrhage in fetal brain in both experimental groups. Fetal liver karyotyping analysis was normal in fetuses of all groups and TUNEL assay in fetal striatum did not show significant difference in number of apoptotic cells between groups.
Conclusion: From our results, it could be concluded that chronic abuse of MA by pregnant females during organogenesis period can cause teratogenic effect and brain hemorrage in fetus.
Roshangar Leila, Seddighe Abdollahifard, Abbas Majdi, Armin Zarrintan, Alia Ghasemzade, Laaia Farzadi, Sara Soleimani Rad, Jafar Soleimani Rad,
Volume 11, Issue 5 (7-2013)

Background: More than 40% of infertilities are due to endometriosis. Ultrustructural and histochemical study of endometrium will help to clarify the etiology of endometriosis.
Objective: The aim of the present study was to investigate the ultrastructure and occurrence of apoptosis in endometrial cells of women with or without endometriosis.
Materials and Methods: In the present case-control study, endometrial specimens from 12 women without endometriosis (as control) and 12 women with endometriosis (as case) were examined. Specimens for control group were obtained from the patients that were referred to gynecology hospital for hysterectomy due to various reasons. In case group the endometriosis was diagnosed according to laparoscopy and endometrial samples were taken using pippel biopsy. The specimens from both case and control groups were processed for Transmission Electron Microscopy (TEM), TUNEL reaction technique and morphometric studies.
Results: The results show that endometrial epithelium lost its continuity in women with endometriosis and endometrial cells have euchromatic nucleus in comparison to those from non-endometriosis. There were several apoptotic cells in the luminal and glandular endometrial epithelium and stroma from endometrium of control group. However, apoptotic cells were rarely seen in the endometrium from women with endometriosis. The difference in number of apoptotic cells between two groups statically was significant (p<0.001).
Conclusion: Regarding the ultrastructural characteristics of endometrial epithelial cells and comparison of apoptotic occurrence in control and case groups it is concluded that endometrial cells in endometriosis group have higher potential to survive and possibly implant.
Farnaz Shapouri, Shaghayegh Saeidi, Sara Ashrafi Kakhki, Omid Pouyan, Elham Amirchaghmaghi, Reza Aflatoonian,
Volume 11, Issue 11 (12-2013)

Background: It has been suggested that malfunction of immune system may cause testicular cancer. Recently, our understanding of innate immune system has been expanded, by discovery of “Toll- like receptors” (TLRs). Some studies have shown that polymorphisms of TLR2 and 4 can effect on the risk of cancer. Also, the role of TLRs 3and 9 have been shown in apoptosis of cancer cells and metastasis in animal models.
Objective: Little information is available about the influence of innate immunity on testicular malignancy. Therefore, expression of TLRs 2, 3, 4 and 9 as main components of innate immunity has been investigated in this study.
Materials and Methods: In this case control study, TLRs gene expression was examined by RT-PCR in normal testis and testicular cancer tissues. Real time quantitative PCR (Q-PCR) analysis was used to compare the relative expression of TLRs between the samples.
Results: mRNAs of TLR 2, 3, 4 and 9 were expressed in all normal and cancer samples. Q-PCR reveals that cancer samples had stronger expression of these genes in compared with normal ones.
Conclusion: It seems that the different TLRs expression in testicular cancer cells may contribute to extensive signaling pathways involved in carcinogenesis.
Wesam Kooti, Esrafil Mansori, Maryam Ghasemiboroon, Mahmoud Harizi, Ashraf Amirzarga,
Volume 12, Issue 5 (6-2014)

Infertility is one of the health problems that will have a negative impact on the individual, social and economic and is seen in 10-15% of couples (1). About 40 % of the issues involved with infertility are due to the man. Male sperm cells count nowadays has decreased dramatically in comparison with those who lived 50 years ago (2). Causes of infertility in men are included: oligozoospermia, immaturity of sperm, sperm deformity, and sperm non-motility. Spermatogenesis takes place within the testes under control of testosterone secreted by the testes and secretory activity of the testes controlled by the hypothalamic-pituitary-testicle axis. Due to adverse effects and side effects of chemical drugs today, the use of traditional medicine, especially herbal therapy is taken into consideration. In traditional medicine, it has been pointed to therapeutic properties of celery. Celery has anti-fungal, anti-bacterial, and anti-cancer properties (3). Also this plant is an appetite stimulant and sexual booster (4). Previous studies have shown that sperm cells are largely vulnerable to oxidative stress but celery is rich in antioxidant compounds such as flavonoids (apiin and apigenin), vitamins E and C that can reduce oxidative stress (5, 6). So, in the present study the protective effect of celery was investigated on the cauda epididymal spermatozoa and testis in rat. A total number of 32 male Wistar rats (weighting 170-220 g) were prepared from animal house central of Ahvaz Jundishapur University of Medical Sciences. Animals were maintained in plastic cages with 12/12 h light/dark cycle at 21±2oC. All experimental animals were carried out in accordance with Ahvaz University Ethical Committee. Hydro-alcoholic extract of celery was prepared by maceration method. The rats were divided into four groups of 8 animals each: control, did not receive anything; vehicle, received propylene glycol; experimental groups, and received hydro-alcoholic extract of celery with doses of 100 and 200 (mg/kg) with solvent of propylene glycol by gavage once every 48 hours for twenty days. At the end of 20th day, rats were scarified under ketamine and xylazine anesthesia then the epididymis and testes were carefully separated. The epididymis was used for sperm count and testes were prepared for morphometric and histologic evaluation. Statistical significance of differences were assessed with one-way ANOVA by SPSS for windows (version 15) followed by LSD test. P&lt;0.05 was assumed as statistically significant. Results of morphometric studies indicated a decrease in number of primary spermatocytes, Sertoli cells and sperm as well as an increase of lumen diameter of seminiferous tubules in vehicle group when compared to the control (p&lt;0.05), but there was not different between the experimental groups and control (p&gt;0.05). Evaluation of tissue sections showed that germinal epithelium in the control group was normal and tissue damage was not observed in epithelial tissue. However, in the vehicle group, epithelium was destroyed and arrangement of epithelial cells was disordered, and fluid aggregation is seen into the epithelial cell and also, reduction of epithelium thickness was observed. In the experimental group (100 mg/kg), there was arrangement in germinal epithelium cells but fluid aggregation was observed into the epithelial cells. A reduced epithelial thickness was seen only in some tubules. However, all these histological changes were less than the vehicle group. In the experimental group (200 mg/kg), tissue destruction was largely improved, and there was an arrangement of the epithelial cells, there was not fluid aggregation into the epithelium, and the thickness of the epithelium was returned almost too normal state. The results of the present study showed that hydro-alcoholic extract of celery improved the destructive effects of propylene glycol on the testes and sexual cells. These findings are similar to previous studied (7, 8). Previous studies have demonstrated that excessive alcohol consumption in men can cause a deficiency in testosterone production and testicular atrophy. Testicular atrophy results primarily from the loss of spermatogenic cells of the seminiferous tubules that this can be caused by oxidative stress generated by alcohol (9). Researches also indicated alcohol with involvement of phase system and activation of caspases induced apoptosis in testicular cells (10). Spermatogenesis and maturation of sexual cells depends on protection of cytotoxic and pathologic lesions that threatens these events. Free radicals due to a strong desire to get electrons induce damage to molecules such as fatty acid of biological membranes and its oxidation. Celery is a strong antioxidant due to flavonoids such as apiein and apigenin (5, 6). Antioxidant compounds are able to protect cell membranes against damage. Antioxidants directly or indirectly impact on hypothalamic-pituitary-testicular axis thus increase sperm count and fertility (5, 6). So celery can be considered as a medicinal herb for infertility. However, further clinical studies are recommended. The study in the form of a research plan was approved with no 91s8 of Research Deputy of Ahvaz Jundishapur University of Medical Sciences. Finally, we acknowledge deputy vice-chancellor for research affairs of AJUMS for financial support, and particularly Research Consultation Center (RCC) for technical support.
Marzieh Rahimipour, Ali Reza Talebi, Morteza Anvari, Abolghasem Abbasi Sarcheshmeh, Marjan Omidi,
Volume 12, Issue 5 (6-2014)

Background: Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans.
Objective: The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice.
Materials and Methods: Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin (0.2% w/v) for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham’s F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology (Pap-staining) and viability (eosin-Y staining). Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD (sperm chromatin dispersion) and terminal deoxynucleotidyl transferase (TUNEL) assay.
Results: Following saccharin consumption, we had a reduction in sperm motility with respect to control animals (p=0.000). In addition, the sperm count diminished (17.70±1.11 in controls vs. 12.80±2.79 in case group, p=0.003) and the rate of sperm normal morphology decreased from 77.00±6.40 in control animals into 63.85±6.81 in saccharin-treated mice (p=0.001). Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one (p=0.001, p=0.002 respectively).
Conclusion: Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice.

Alireza Abhari, Nosratollah Zarghami, Laya Farzadi, Mohammad Nouri, Vahideh Shahnazi,
Volume 12, Issue 10 (11-2014)

Background: MicroRNA (miRNA) is small endogenous, single strand RNA molecules that regulate gene expression at post-transcriptional level through several mechanisms to affect key cellular event including male germ cells differentiation, proliferation, development and apoptosis. Mutation and/or aberrant expression of miRNAs have been associated with progression of various disorders, including infertility.
Objective: The purpose of this research was to study the estrogen receptor beta (ERβ), hsa-mir-21 and, hsa-mir-22 expression level in oligospermic infertile and control fertile men and correlation between them.
Materials and Methods: In this study, the change in mir-21, mir-22 expression and their common target gene (ERβ) expression levels were evaluated in oligospermic infertile men (n= 43) compared with 43 age matched healthy control by Real-Time PCR methods.
Results: Expression analysis by qRT-PCR test on miRNA have identified that mir-21, mir-22 levels were significantly higher than those in normal controls (p<0.0001) and ERβ expression level significantly decreased in comparison with the normal group (p<0.0001).
Conclusion: Our study showed that mir-21 and mir-22 are indirectly involved in spermatogenesis by regulating of the estrogen receptor and might have a diagnostic and prognostic value in men infertility.
Behrooz Niknafs, Ahmad Mehdipour, Amaneh Mohammadi Roushandeh,
Volume 12, Issue 12 (12-2014)

Background: Melatonin, a reactive oxygen species (ROS) scavenger and an antioxidant, has been shown that can inhibit apoptosis. Administration of melatonin may improve embryo development in assisted reproductive technology (ART).
Objective: The aim of this study was to evaluate the role of melatonin in inhibition of spontaneous and induced apoptosis by Tumor Necrosis Factor Alph (TNF-α) and actinomycin-D during preimplantation development of mouse embryos.
Materials and Methods: Female BALB/c mice were superovulated with pregnant mare serum gonadotropin (PMSG) followed by human chorionic gonadotropin (HCG), then allowed to mate with male mice. The resultant 2-cell embryos were divided into six groups as follows: control (group I), melatonin (group II), actinomycin-D (group III), actinomycin-D + melatonin (group IV), TNF-α (group V), and TNF-α + melatonin (group VI). We recorded the numbers and developmental rates of the 4-cell, 8-cell, morula and blastocyst embryos. Blastocysts were stained with acridine orange in order to assess for the embryo quality.
Results: The group IV showed a significantly higher developmental rate of blastocysts compared to group III (p<0.05). The number of dead blastomers was significantly decreased in group IV in comparison to group III (p<0.05). Both V and VI groups had a lower developmental rate and lesser quality of blastocysts compared with group I. There was no significant difference in the developmental rate of blastocysts from group II compared to group I (p<0.05).
Conclusion: Supplementation of embryo culture media with melatonin can improve the quality and developmental rate of embryos. Melatonin can prevent cell death that was induced by TNF- α and actinomycine-D.
Arezoo Khoradmehr, Amirhossein Danafar, Iman Halvaei, Jalal Golzadeh, Mahya Hosseini, Tahereh Mirjalili, Morteza Anvari,
Volume 13, Issue 1 (1-2015)

Background: Methamphetamine (MA) is one of most common illicit drugs which were reported that nearly half of MA consumers are women. MA can cross through placenta and affects pregnancy and fetus development.
Objective: Our aim was to evaluate effects of injected MA on crown-rump length, head and placental circumference, body weight, histological changes and apoptosis in fetus.
Materials and Methods: Twenty-four NMRI pregnant mice were randomly divided into five groups. First, second and third groups were injected intraperitoneally 10 mg/kg/day MA during gestational days (GD): GD1-7, GD8-14, and GD1-14, respectively. Forth group, as sham, was injected saline from GD1-14, and finally control which was received neither MA nor saline. On GD15 cervical dislocated pregnant mice, fetus and placenta were weighed and fetus crown-rump length was measured. Hematoxylin and Eosin staining and TUNEL assay were applied to assess histological changes and apoptosis respectively.
Results: Fetus body weight and crown-rump length showed significant decrease in third compared to first and second groups (p≤0.001). There were significant differences in head circumference in control and sham compared to third group (0.5 (0.5-0.6), 0.6 (0.5-0.8), 0.4 (0.4-0.5) cm respectively, p≤0.001). Also fetus that treated with MA showed lower placenta circumference compared to control and sham groups. Histological changes such as exencephaly, hemorrhage and immature fetus were observed in second and third groups. Apoptotic cells in second and third groups were higher than controls, but differences were not significant.
Conclusion: It seems MA abuse during pregnancy can cause morphological and histological changes in mice fetus but the exact mechanism remains unclear.
Ali Shalizar Jalali, Gholamreza Najafi, Mohammadreza Hosseinchi, Ashkan Sedighnia,
Volume 13, Issue 1 (1-2015)

Background: Stanozolol (ST) is a synthetic anabolic-androgenic steroid often abused by athletes. An increasing body of evidence points towards the role of ST misuses in the pathogenesis of a wide range of adverse effects including reprotoxicity.
Objective: The aim of this study was to analyze the possible reproprotective effect of royal jelly (RJ) as an efficient antioxidant in ST-treated mice.
Materials and Methods: Adult male mice were divided into four groups (n=5). Two groups of mice received ST (4.6 mg/kg/day) via gavage for 35 days. RJ was given orally to one of these groups at the dose level of 100 mg/kg body weight per day synchronously. Untreated control group and RJ-only treated group were also included. Epididymal sperm characteristics and in vitro fertilizing capacity were evaluated after 35 days.
Results: ST treatment caused a significant (p<0.05) decrease in sperm count and motility and fertilization rate along with poor blastocyst formation and increased sperm DNA damage. Moreover, the incidence of apoptosis and abnormality in spermatozoa was significantly (p<0.05) higher in ST-exposed mice than those of control. The above-mentioned parameters were restored to near normal level by RJ co-administration.
Conclusion: Data from the current study suggest that RJ has a potential repro-protective action against ST-induced reproductive toxicity in mice. However, clinical studies are warranted to investigate such an effect in human subjects.
Majid Taati, Mehrnoush Moghadasi, Omid Dezfoulian, Payman Asadian, Morteza Zendehdel,
Volume 13, Issue 2 (2-2015)

Background: Testicular torsion is a medical emergency that requires surgical intervention to reperfuse the affected testis. Ischemia reperfusion injury is usually associated with proinflammatory cytokine generation and apoptosis of germ cells in the testes.
Objective: In this study we investigate the effect of ghrelin on the proinflammatory cytokines levels and germ cell apoptosis in testicular ischemia reperfusion.
Materials and Methods: 45 male rats were selected for the study and randomly divided into 3 groups, each containing 15 rats. Animals in the testicular torsion and ghrelin treated groups were subjected to unilateral 720 counterclockwise testicular torsion for 1 hr and then reperfusion was allowed after detorsion for 4 hr, 1 and 7 days. The ghrelin-treated group received intraperitoneal injection of ghrelin 15min before detorsion. The expression levels of bcl-2-associated X protein and proliferating cell nuclear antigen in testicular tissue in the different groups were detected by immunohistochemical assay and tissue cytokines interleukin-1β, tumor necroses factor-α and interleukin-6 were measured using enzyme-linked immunosorbent assay
Results: After being treated by ghrelin, the population of immunoreactive cells against BAX in the spermatocytes on day 7 after reperfusion significantly decreased when compared to tortion/ detortion-saline animals (p=0.024). In contrast, PCNA expression in the spermatocytes and spermatogonia were not significantly different between tortion/ detortion-ghrelin and tortion/ detortion-saline groups on both experimental days. Administration of ghrelin significantly attenuated the testicular tumor necroses factor-α and interleukin-6 levels compared with the untreated animals, but had no significant effect on the level of interleukin-1β.
Conclusion: Ghrelin offers remarkable anti-inflammatory and anti-apoptotic effects in testicular ischemia reperfusion injury.
Arash Khaki,
Volume 13, Issue 3 (3-2015)

Background: Antibiotic therapies used in treatment of many diseases have adverse effects on fertility. This review analyzes previous comparative studies that surveyed the effects of two common groups of antibiotics on male fertility.
Objective: To evaluate histo-pathological effects of fluoroquinolones and aminoglycosides on sperm parameters and male reproductive tissue.
Materials and Methods: Articles about the effects of aminoglycosides and fluoroquinolones on male infertility, sperm parameters, male reproductive tissue, and spermatogenesis in English and Persian languages published on Google Scholar and PubMed databases from January 2000 to December 2013 were assessed. Randomized controlled trials (RCTs) assessing the effects of aminoglycosides or fluoroquinolones on sperm parameters, artificial insemination, and male reproductive tract or RCTs comparing aminoglycosides vs. fluoroquinolones were eligible for inclusion. For ascertaining the reliability of study, data were extracted independently and in duplicate by two investigators.
Results: Sperm viability was decreased significantly with streptomycin, gentamicin, and neomycin (p<0.001). Sperm motility was decreased significantly with gentamicin and neomycin (p<0.05). Total sperm count was significantly decreased with ofloxacin, gentamicin, streptomycin, and neomycin (p<0.022). There was significant decrease in post-thawing motility with low dose and high dose of ciprofloxacin. Testis weight was decreased with gentamicin and ofloxacin significantly (p<0.011). There was significant decrease in seminal vesicle weight with gentamicin, neomycin, and ofloxacin (p<0.022). Furthermore, changes in epididymis weight, percentage of total apoptotic cells, and diameter of seminiferous tubule were significant with all drugs including streptomycin, gentamicin, neomycin, and ofloxacin (p<0.05).
Conclusion: Streptomycin has less negative effects on cell’s apoptosis and sperm parameters as compared to other drugs. Gentamicin has more detrimental effects so lesser dosage and duration is recommended. Fluoroquinolones showed negative effects on testis tissue and sperm parameters. Ciprofloxacin has less adverse effects than gentamicin in artificial insemination.

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