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

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

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

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

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

Cytochrome C and caspase-3/7 are involved in mycophenolic acid-induced apoptosis in genetically engineered PC12 neuronal cells expressing the p53 gene

Mycophenolic acid [MPA] is the active metabolite of mycophenolate mofetil. This study designed to investigate the mechanism of cytotoxicity of MPA on the genetically engineered PC12 Tet Off [PTO] neuronal cells with p53 gene. Alamar Blue [AB] reduction showed concentration-dependent cytotoxicity of MPA on PTO cells with IC50 value of 32.32 +/- 4.61 mM. The reactive oxygen species [ROS] generation following exposing the cells to MPA showed a significant [p < 0.05] increase in the ROS production and in a concentration-dependent fashion. Involvement of Caspase 3/7 proteases and Cytochrome C release in the induction of DNA fragmentation are all hallmarks of MPA-induced apoptosis in PTO cells. Our data suggest that MPA exerts an apoptotic effect on PTO cells. Moreover, the apoptotic effect of MPA attribute to the elevation of ROS generation by which might trigger the cytochrome C release and the activation of Caspase 3/7 that ultimately results in DNA fragmentation

Aflatoxin M1 concentration in various dairy products: evidence for biologically reduced amount of AFM1 in Yoghurt

Aflatoxin M[1] [AFM[1]], a carcinogenic substance is found in milk and dairy products. The effect of season and type of dairy products on AFM[1] level in northern Iran was investigated in this study. Three hundred samples [each season 75 samples] including raw and pasteurized milk, yoghurt, cheese, and cream samples were collected from three distinct milk producing farms. The samples were subjected to chemical and solid phase extractions and were analyzed by using HPLC technique. Recovery percentages, limit of detection and limit of quantification values were determined. Seventy percent and 98% were the minimum and maximum recoveries for cheese and raw milk, respectively and 0.021 and 0.063 ppb were the limit of detection and limit of quantification values for AFM[1]. We found that in autumn and winter the highest level [0.121 ppb] of AFM[1] in cheese and cream samples and failed to detect any AFM[1] in spring samples. Interestingly, our data showed that the yoghurt samples had the lowest level of AFM[1] in all seasons. There are significant differences between the AFM[1] levels in dairy products in various seasons and also various types of products, suggesting spring and summer yoghurt samples as the safest products from AFM[1] level point of view

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

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

Crataegus monogyna aqueous extract ameliorates cyclophosphamide-induced toxicity in rat testis: stereological evidences

Cyclophosphamide [CP] is extensively used as an antineoplastic agent for the treatment of various cancers, as well as an immunosuppressive agent. However, despite its wide spectrum of clinical uses, CP is known to cause several adverse effects including reproductive toxicity. Crataegus monogyna is one of the oldest pharmaceutical plants that have been shown to be cytoprotective by scavenging free radicals. The present study was conducted to assess whether Crataegus monogyna fruits aqueous extract with anti-oxidant properties, could serve as a protective agent against reproductive toxicity during CP treatment in a rat model. Male Wistar rats were categorized into four groups. Two groups of rats were administered CP at a dose of 5 mg in 5 ml saline/kg/day for 28 days by oral gavages. One of these groups received Crataegus monogyna aqueous extract at a dose of 20 mg/kg/day orally four hours after cyclophosphamide administration. A vehicle treated control group and a Crataegus monogyna control group were also included. The CP-treated group showed significant decreases in the body, testes and epididymides weights as well as many histological alterations. Stereological parameters and spermatogenic activities [Sertoli cell, repopulation and miotic indices] were also significantly decreased by CP treatment. Notably, Crataegus coadministration caused a partial recovery in above-mentined parameters. These findings indicate that Crataegus monogyna may be partially protective against CP-induced testicular toxicity

Testicular biohistochemical alterations following experimental varicocele in rats

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

cytotoxicity and comparative antibacterial study on the effect of Zataria multiflora boiss, trachyspermum copticum essential oils, and enrofloxacin on aeromonas hydrophila

This study designed to test the antibacterial potency of enrofloxacin [ENR] and essential oils from Zataria multiflora Boiss [ZEO] and Trachyspermum copticum [TEO] on Aeromonas hydrophila. The antibacterial potency of test compounds was determined by several methods including the inhibition zone diameter determination, microbroth dilution method and colorimetric method of MTT. The cytotoxicity of test substances was assessed on Chinook salmon [Oncorhynchus tshawytscha] embryo [CHSE-214] cells. Results showed that ENR and tested essential oils exert antibacterial effect against A. hydrophila. Moreover, ENR exerted the most potent antibacterial effect with MIC values of 62.5 ng/ml. The natural compounds of ZEO and TEO also showed antibacterial effects with rather high MIC values of 0.315 mg/ml, and 1.25 mg/ml, respectively. None of the tested substances showed toxicity on CHSE-24 cells. It is concluded that ZEO and TEO could be applied to prevent from A. hydrophila infection. Moreover, data also suggest that MTT method could be both cost- and time-effective and accurate method of MIC determination

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

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

effect of tamoxifen on the fetal and neonatal ovarian follicles development in rats

The effect of tamoxifen as a selective estrogen receptor modulator which is widely used for treatment of early and metastatic breast cancer was investigated on the folliculogenesis in rat's fetuses and neonates. The pregnant rats assigned into test and control groups. Control group received olive oil and treatment groups received either 17-[3-estradiol [10 [micro g/kg/day] or tamoxifen [0.4 mg/kg/day] between days 8-13 of pregnancy. On day 20th of pregnancy the rats euthanized and the blood samples were collected for determination of FSH, E[2], and the fetuses fixed for histological studies. Another group of pregnant rats went forward to obtain their neonates and we euthanized the neonates and the genital system was collected for further histopathological analyses on day 5th. The histological examinations of the fetus's and neonate's ovaries and biochemical data showed significant changes in the rats which treated with tamoxifen. The absence of folliculogenesis and an increase in E[2] level in tamoxifen-treated group which accompanied with sharp decrease of FSH level in comparison with the control group were demonstrated. By contrary, E[2] treated group showed a positive progress in development in terms of the formation of secondary follicles and also supportive connective tissues in comparison with the control group. In conclusion, this study supports the previous findings showing that tamoxifen has effects on the development of ovaries and therefore, it should be avoided or used with great caution in pregnant women

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

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