Protective effect of aqueous and methanolic extracts of corn silk on nicotine-induced reproductive system disorders in male mice.

OBJECTIVE: The protective effect of aqueous and methanolic extracts of corn silk on reproductive disorders induced by nicotine was investigated in the present study. METHODS: In this experimental study, 30 male NMRI mice (25-30gr) were divided into 5 groups: controls, sham, nicotine 2.5mg/kg, nicotine+aqueous extract of corn silk 400mg/kg, and nicotine+methanolic extract of corn silk 400mg/kg for 34 days. One day after the last nicotine and extracts administration, the serum samples were collected through cardiac puncture for hormonal measurements, and the testis and tail of the epididymis were isolated for the testis antioxidant, morphology, histopathology assessments, and sperm count. RESULTS: Luteinizing hormone (LH) and malondialdehyde (MDA) increased in the nicotine group. Testosterone, sperm count, and glutathione (GSH) decreased when compared to the control group. Both aqueous and methanolic extracts of corn silk led to the improvement of mentioned changes; Except for GSH, because only treatment with methanolic extract could lead to its increase (p<0.05). Nicotine decreased the thickness of the epithelium of seminiferous tubules and the separation between them, and the administration of corn silk extracts improved that. CONCLUSIONS: Nicotine consumption increased oxidative stress, LH levels, and decreased testosterone and sperm count, which indicate the induction of primary hypogonadism in animals. Moreover, the use of corn silk extracts has recovered the amounts of sex hormones and sperm count to normal conditions by reducing lipid peroxidation.

The nitric oxide-cyclic GMP-KATP channels pathway contributes to the effects of montelukast against gastric damage induced by ethanol.

The leukotrienes, lipid mediators, have a role in gastric damage induced by ethanol. Here, the gastroprotective effect of montelukast, an antagonist of the leukotriene receptor, and the involvement of the NO-cGMP-KATP channel pathway, were evaluated in gastric damage induced by ethanol in rats. For this, l-arginine, l-NAME, methylene blue (guanylate cyclase inhibitor), sildenafil, diazoxide, or glibenclamide (ATP-sensitive potassium channel blocker) were administered 30 min before montelukast (0.1, 1, 10, and 20 mg/kg, by mouth [p.o.]). After 1 h, to induce gastric damage, the rats received absolute ethanol (4 mL/kg, p.o.), and then microscopic, macroscopic, and pro-inflammatory parameters (TNF-α and IL-1ß) were assessed. Results obtained here revealed that montelukast significantly attenuated the macroscopic and microscopic lesions induced by ethanol. Montelukast also reduced IL-1ß and TNF-α levels. It was also observed that NOS inhibitor (l-NAME), methylene blue, and glibenclamide inhibited the effects of montelukast in the stomach. Moreover, the NO precursor (l-arginine), the PDE-5 inhibitor (sildenafil), and a potassium channel opener (diazoxide) before montelukast produced gastroprotective effects. In conclusion, the effect of montelukast against gastric lesions induced by ethanol is mediated, at least in part, through the pathway of the NO-cGMP-KATP channel.

Effects of aqueous and hydro-alcoholic extracts of Seidlitzia rosmarinus on male mouse reproductive system: An experimental study.

OBJECTIVE: This study investigated the effects of aqueous and hydro-alcoholic extracts of Seidlitzia rosmarinus on reproductive hormones, sperm variables, and antioxidant enzymes level in the mice testis. METHODS: In this experimental study, 24 three-month-old male NMRI mice weighing (25-30g) were divided into three groups: control, aqueous and hydro-alcoholic extracts of Seidlitzia rosmarinus 100mg/kg. Dissolved extracts were gavaged orally for 35 days. One day after receiving the last dose of the extract, the blood sample, testis, and the epididymis tail were taken for plasma hormonal, testicular antioxidants level, sperm count, and vitality assessments. RESULTS: Testicular level of malondialdehyde increased in aqueous and hydro-alcoholic extracts groups (p=0.04); total antioxidant capacity decreased in aqueous and hydro-alcoholic extracts groups (p=0.008); and the consumption of aqueous (p<0.001) and hydro-alcoholic (p=0.03) extracts decreased catalase in comparison with the control group. The plasma level of luteinizing hormone decreased in the aqueous extracts administrated group (p=0.009); the follicle-stimulating hormone increased in aqueous (p=0.03), and hydro-alcoholic extracts administered mice; and the testosterone level decreased in aqueous extract-treated animals versus the control group (p<0.001). The sperm count was increased in aqueous (p=0.04) and hydro-alcoholic (p=0.009) extracts groups, but its vitality was decreased (p=0.008) in comparison with the control group. CONCLUSIONS: In conclusion, Seidlitzia rosmarinus has an adverse effect on male reproductive hormones and sperm viability via increased lipid peroxidation and reduced antioxidant defense system performance.

Anesthesia and analgesia for common research models of adult mice.

Anesthesia and analgesia are major components of many interventional studies on laboratory animals. However, various studies have shown improper reporting or use of anesthetics/analgesics in research proposals and published articles. In many cases, it seems "anesthesia" and "analgesia" are used interchangeably, while they are referring to two different concepts. Not only this is an unethical practice, but also it may be one of the reasons for the proven suboptimal quality of many animal researches. This is a widespread problem among investigations on various species of animals. However, it could be imagined that it may be more prevalent for the most common species of laboratory animals, such as the laboratory mice. In this review, proper anesthetic/analgesic methods for routine procedures on laboratory mice are discussed. We considered the available literature and critically reviewed their anesthetic/analgesic methods. Detailed dosing and pharmacological information for the relevant drugs are provided and some of the drugs' side effects are discussed. This paper provides the necessary data for an informed choice of anesthetic/analgesic methods in some routine procedures on laboratory mice.

Comparison of Follicular Fluid Paraoxonase 3 Level, Ovarian Hormones and Oocyte Quality between Fertile and Infertile Women.

Background: The purpose of the current study was to evaluate the possible effect of follicular fluid paraoxonase 3 (PON 3) on oocyte quality and sex hormones. Methods: This descriptive-analytical study was performed on totally 90 enrolled women including fifty infertile women presenting with polycystic ovaries and uni-lateral tubal factor and forty fertile women with male factor infertility referring to Umm-al-Banin Infertility Clinic in Dezful, Iran for in vitro fertilization during October 2018 to November 2019. Oocyte removal was carried out under transvaginal ultrasound guidance, and follicular fluid (FF) was removed and preserved to detect PON3, estrogen, and progesterone levels. In addition, oocyte number and quality were assessed and its association with PON3 activity in the FF was evaluated. Oneway ANOVA and Fisher's least significant difference (LSD) were used for data analysis and p≤0.05 were considered statistically significant. Results: A significant increase was observed in the total number of the oocytes and mature metaphase II oocytes with ≥20 pg/ml of PON3 concentration in the FF (p≤0.05). Moreover, a positive relationship was shown between the increased estradiol level in follicular fluid and PON3, so that the highest estradiol level was observed in the amount of 31-40 pg/ml of PON3 (p≤0.05). Conclusion: According to the results, as the number of the mature oocytes increased, the amount of PON3 as well as estradiol levels in the FF increased. This research displays an increase in the level of PON3 with mature oocytes, thus supporting the indirect evidence for the function of PON3 in follicle development.

The effect of montelukast, a leukotriene receptor antagonist, on the acetic acid-induced model of colitis in rats: Involvement of NO-cGMP-KATP channels pathway.

Inflammatory bowel disease is a chronic autoimmune disorder that may involve entire gastrointestinal tract. The leukotrienes have a role as mediators in the pathophysiology of colitis. Here, we investigated the effect of a leukotriene receptor antagonist, montelukast, and also the role of the NO-cGMP-KATP channel pathway in acetic acid-induced colitis. Rectal administration of acetic acid (4%) was used for induction of colitis in rats. To investigate our hypothesis, the rats were intraperitoneally pre-treated with L-NAME (NOS inhibitor), L-arginine, sildenafil, methylene blue, glibenclamide, or diazoxide 15 min before treatment with montelukast (5-20 mg/kg, i. p.), for three consecutive days. Then, microscopic, macroscopic, and inflammatory parameters were evaluated. Montelukast reduced the microscopic and macroscopic damage induced by acetic acid. Montelukast also reduced the level of IL-1ß and TNF-α. We also showed that the effects of montelukast were significantly attenuated by L-NAME, methylene blue (guanylate cyclase inhibitor), and an ATP-sensitive potassium channel blocker (glibenclamide). Also, the administration of L-arginine, sildenafil, and diazoxide before montelukast produced protective effect. In conclusion, the pathway of the NO-cGMP-KATP channel is involved in the protective effect of montelukast in acetic acid-induced colonic tissue damage.

Changes of Serum Level of Homocysteine and Oxidative Stress Markers by Metformin and Inositol in Infertile Women with Polycystic Ovary Syndrome: A Double Blind Randomized Clinical Trial Study.

Background: Hyperhomocysteinemia plays an important role in the anovulation in infertile women suffering from polycystic ovary syndrome (PCOS). However, long-term metformin therapy elevated homocysteine (Hcy) concentration in these individuals. Inositol increases serum insulin levels and improves ovulation. The aim of this study was to compare the effect of metformin and inofolic on the level of serum Hcy and oxidative markers in the infertile patients with PCOS. Materials and Methods: Eighty PCOS infertile women undergoing in vitro fertilization in the Umm-al-Banin clinical center, Dezful, Iran from December 2018 to September 2019 were invited to participate in this double blind randomized clinical trial. They were divided into two groups; group A who received metformin (1000 mg twice/day) and folic acid (400 µg /day) and group B who used inofolic (inositol+ olic acid 200 µg twice/day) for 3 months. Results: The mean Hcy levels increased significantly by metformin (P=0.02), but not by inofolic. There was a decrease in the total antioxidant capacity (TAC) after metformin administration (P=0.01). In both groups, a significant increase in folic acid levels was observed after treatment (P=0.04). Also, no significant change in vitamin B12 and malondialdehyde levels was observed in both groups (P=0.08). Conclusion: These findings indicate an increase in the serum Hcy levels as well as a remarkable decrease in TAC following metformin treatment. Given the rise in blood Hcy in PCOS patients, inofolic and other medications containing inositol can be prescribed instead of metformin (registration number: IRCT20190508043516N1).

The effect of follicular fluid selenium concentration on oocyte maturation in women with polycystic ovary syndrome undergoing in vitro fertilization/Intracytoplasmic sperm injection: A cross-sectional study.

BACKGROUND: A high level of free radicals and oxidative substances in women with polycystic ovary syndrome (PCOS) can affect the ovaries through oxidative stress. Antioxidants such as selenium, a vital trace element in human health, can improve the prognosis of PCOS by reducing oxidative stress. OBJECTIVE: This study was performed due to the lack of comprehensive information about selenium concentration in follicular fluid and its effect on the oocyte count and quality in infertile women with PCOS. MATERIALS AND METHODS: In this cross-sectional study, 78 women with PCOS referred to Umm-al-Banin Infertility Clinic Center, Ganjavian Hospital, Dezful, Iran for in-vitro fertilization from March to November 2019 were enrolled. After ovarian stimulation with the antagonist protocol, the oocytes were retrieved under transvaginal ultrasound in in-vitro fertilization/intracytoplasmic sperm injection cycles, and selenium concentrations were measured in the follicular fluid using an atomic absorption method by spectrophotometer device. Oocyte count and morphology were evaluated using inverted optical microscopy. RESULTS: There were no significant differences between follicular fluid selenium concentrations in terms of the total number of oocytes and immature oocytes in the metaphase I and germinal vesicle stages. However, a significantly reduced number of metaphase II oocytes was observed at selenium levels < 40 µg/dL (p = 0.001). CONCLUSION: Based on our results, low levels of follicular selenium concentration in infertile women with PCOS can reduce the quality and potency of oocyte maturation.

Follicular Fluid Zinc Level and Oocyte Maturity and Embryo Quality in Women with Polycystic Ovary Syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is considered to be one of the most common endocrine disorders in women of reproductive age. Zinc, a vital trace element in the body, plays a key role in maintaining health, especially due to its antioxidant role. On the other hand, lack of antioxidants and oxidative stress can adversely affect oocytes quality and consequently fertility rate. The available studiesthat report the effect of follicular fluid (FF) zinc in terms of the number and quality of the oocytes in infertile women with PCOS, are few and not consistent. We decided to investigate this issue. MATERIALS AND METHODS: In this cross-sectional study, from the women with PCOS referring to Omolbanin Hospital, Dezful, Iran (February to December 2019), a total of 90 samples (follicular fluid, oocytes, and embryos) were collected from those who had undergone in vitro fertilization (IVF). To measure zinc level in follicular fluid, high performance liquid chromatograpy (HPLC) was utilized. Also, oocytes maturity and embryos quality evaluation was performed using inverted optical microscopy. One-way ANOVA and Fisher's least significant difference (LSD) were used for data analysis. RESULTS: The amount of FF zinc was not associated with any significant differences in the number of oocytes and metaphase I (MI) and germinal vesicle (GV) oocytes, but a significant decrease was observed in the number of metaphase II (MII) oocytes at zinc values lessthan 35 µg/dL. The FF zinc levels lessthan 35 µg/dL were also significantly associated with decreased embryo quality. CONCLUSION: A significant relationship was found between the level of FF zinc and the quality and the number of oocytes taken from the ovaries of infertile patients with PCOS history who were candidates for IVF treatment as well as the number of high quality embryos.

Evaluation of the Effect of Captopril and Losartan on Tacrolimus-induced Nephrotoxicity in Rats.

BACKGROUND: Tacrolimus is an immunosuppressive drug. Activation of the renin-angiotensin system (RAS) and associated inflammations may exacerbate the toxic effects of tacrolimus. Given the significant role of the kidney in RAS this study aimed to evaluate the effect of captopril as an angiotensin-converting enzyme (ACE) blocker and losartan as an angiotensin II receptor blocker on tacrolimus-induced nephrotoxicity. MATERIALS AND METHODS: In total, 36 adult male rats weighing 200-250 gr were completely randomized and divided into six groups (control, tacrolimus, tacrolimus and losartan, tacrolimus and captopril, losartan, and captopril) for 30 days. Afterwards, blood urea nitrogen (BUN), creatinine (Cr) and ACE2 enzyme were measured. Also, both kidneys were collected for histological examinations. RESULTS: The level of BUN and Cr significantly increased in tacrolimus group. The level of BUN and Cr were lower in the groups treated with a combination of tacrolimus and losartan or captopril. While ACE2 level increased in the groups receiving a combination of tacrolimus and losartan or captopril, the level of increase was insignificant, compared to the group treated with tacrolimus alone. The glomerulus diameter and the thickness of the renal proximal tubular epithelium significantly decreased in the group treated with tacrolimus alone. the mentioned variables increased in the group treated with a combination of tacrolimus and losartan or captopril, compared to the tacrolimus group. CONCLUSION: According to this study, tacrolimus increased the BUN and Cr levels while decreasing the ACE2 levels. However, tacrolimus in combination with losartan or captopril seemed to decrease the nephrotoxicity of the drug.

Pistacia atlantica's effect on ovary damage and oxidative stress in streptozotocin-induced diabetic rats.

OBJECTIVE: Diabetes mellitus (DM) is associated with numerous complications, including gonadal dysfunction. There are specific traditional medicine remedies for DM, including medicinal herbs. Our study aimed to evaluate the role of Pistacia atlantica's extract in the protection against ovary damage by streptozotocin (STZ)-induced DM in rats. METHODS: We ran this experimental study on 40 adult female Wistar rats. We divided the animals into five groups, control (A); DM (STZ by 60 mg/kg- intraperitoneally) (B); DM + hexane extract of P. atlantica (200 mg/kg -orally) (C); P. atlantica extract (D) and DM + glibenclamide (200 mg/kg -orally) (F). The experiment continued for four weeks, and we administered the extract daily. After slaughtering the rats, we removed the ovaries. We assessed parameters, such as blood glucose and levels of oxidative stress markers as well as histological ovary structure. RESULTS: Blood glucose, malondialdehyde (MDA) levels, and the number of atretic follicles were elevated; catalase (CAT), superoxide dismutase (SOD) levels and the number of corpora lutea were significantly decreased in the untreated diabetic rats. These changes returned to normal or diminished with P. atlantica extract and glibenclamide in the treated rats. CONCLUSIONS: The extract of P. atlantica has antihyperglycemic and antioxidative properties, and it decreased ovarian complications in experimental diabetes mellitus.

Therapeutic effects of mesenchymal stem cells on cutaneous leishmaniasis lesions caused by Leishmania major.

OBJECTIVES: Leishmania major (L. major) is a cutaneous leishmaniasis causative agent. Current chemotherapeutic methods are not totally effective in treatment of this disease. The immunomodulation and tissue repairing capability of mesenchymal stem cells (MSCs), ease of isolation, detection and in vitro culture, have encouraged biologists to use MSCs for cell therapy in different infections such as cutaneous leishmaniasis. METHODS: BALB/c mice (6-8 weeks old) were infected with L. major then divided into four groups and treated with MSCs, Glucantime, Glucantime + MSCs, or PBS. Regression of lesions, potency of macrophages for phagocytosis, proliferation of immune cells against Leishmania soluble antigen, reduction of spleen parasite burden and healing of the lesions were evaluated on days 10, 20 and 30 of treatment. RESULTS: The results indicated that the mice intralesionally injected with MSCs showed significant regression in the lesions produced by L. major by day 30. Proliferation of splenocytes stimulated with SLA (soluble leishmania antigen) in vitro in MSC-treated mice on day 20 was significantly higher than in the other groups. The potency of phagocytosis in macrophages of mice treated with MSCs was significantly higher by day 30 and healing of the lesions in this group of mice showed more progress on histopathological examinations. Spleen parasite burden showed significant reduction in the mice treated with Glucantime + MSCs by day 30. CONCLUSIONS: The results showed that including MSCs in treatment of cutaneous leishmaniasis caused by L. major is a promising approach.

Level of Bisphenol A in Follicular Fluid and Serum and Oocyte Morphology in Patients Undergoing IVF Treatment.

Objective: To assess the correlation between the levels of BPA in the serum and follicular fluid (FF) using oocyte morphology. Materials and methods: In this cross-sectional research, oocyte, FF, and serum samples were obtained from a sample population consisting of 90 women undergone in vitro fertilization in Ganjavian Hospital in Dezful, Iran during October 2017-March 2018. The ELISA kit was utilized for the measurement of the BPA levels. In addition, oocyte morphology simultaneous with inverted optical microscopy. Results: Follicular fluid BPA levels had no significant effect on MII oocytes (p ≥ 0.05). However, the mean levels of degenerated oocytes and germinal vesicle (GV) were significantly higher in the women with high BPA levels in the FF (p ≤ 0.05). Moreover, the mean counts of MII oocytes and oocytes were significantly higher in the women with serum BPA levels of ≤ 50 ng/ml (p ≤ 0.05), while the mean count of GV oocytes was significantly higher in the women with serum BPA levels of ≥ 150 ng/ml (p ≤ 0.05). Conclusion: According to the results, higher FF BPA levels were associated with the higher counts of GVs and oocytes, while oocytes with higher maturity can be achieved in lower levels of BPA in the serum of patients.

Protective effects of Asparagus officinalis extract against Bisphenol A- induced toxicity in Wistar rats.

Asparagus officinalis is an herbal medicine with remarkable antioxidant, anti-inflammatory, and antihepatotoxic properties. The present study aimed to investigate whether Bisphenol A (BPA) could induce histopathological changes and oxidative stress in the liver and kidney tissues of male rats. In addition, we evaluated whether the co-administration of the Asparagus officinalis extract (AOE) could prevent the possible damages. In total, 40 adult male Wistar rats with the mean weight of 200 ±â€¯20 grams were divided into five groups. Control subjects were placed in the first group, while group two was the vehicle (5 ml/kg bwt/day). Subjects in group three were administered with 400 mg/kg of AOE (bwt/day), group four received 10 mg/kg of BPA (bwt/day) dissolved in 5 ml/kg of olive oil, and group five received oral BPA and AOE daily for eight weeks. After the experiments, the blood, liver, and kidneys of the animals were collected and examined. Biochemical results showed a significant elevation in the levels of liver and kidney biomarkers in the BPA group (P ≤ 0.05). Moreover, malondialdehyde was observed to increase, while thiol protein and total antioxidant capacity decreased. Histopathological results of the BPA group indicated dilated and congested central and portal veins and inflammatory areas in the liver. In addition, renal test results showed casts in the urinary tubules and acute tubular necrosis. According to the results, the co-administration of AOE and BPA could increase the total antioxidative capability, thereby improving the function and structure of the liver and kidney tissues. Therefore, AOE is a potential protective agent against oxidative stress, liver, and kidney damage.

Protective Effect of Aloe vera Extract against Bisphenol A Induced Testicular Toxicity in Wistar Rats.

OBJECTIVES: Bisphenol A (BPA), an endocrine-disrupting chemical, has been considered as a possible risk factor for fertility because it induces testicular toxicity. Thus, we sought to analyze the effect of Aloe vera as plant with antioxidant properties on tissues and oxidative stress parameters in male rats. MATERIALS AND METHODS: In this experimental study, 50 adult male Wistar rats (200 ± 20 g) have been used in this 56 day study. Animals were completely randomized and divided into five groups: A1 (control), A2 (vehicle control), A3 (Aloe vera gel 300 mg/kg), B1 (BPA 20 µg/kg bw) and B2 (Aloe vera gel+ BPA). At the end of the study, the rats were anesthetized and 2 ml blood samples were obtained for evaluation of oxidative stress markers. Also, both testes were collected for histological examinations. RESULTS: BPA significantly decreased (P<0.05) body and testis weights. Seminiferous tubule diameter (STD) and height of seminiferous epithelium (HSE), were significantly decreased (P<0.05) in the groups receiving BPA as compared to the control. There was also a reduction in the quantity of spermatocyte and spermatids. Moreover, malondialdehyde (MDA) increased and thiol protein (G-SH) decreased. But, co-administration of Aloe vera with BPA accelerated the total antioxidant capacity and testicular tissue structure healing. CONCLUSIONS: According to our findings, Aloe vera gel extract can overcome the damaging effects of BPA on the reproductive system of rats and protects rats' testes against BPA-induced toxicity.

Topical vitamin K1 promotes repair of full thickness wound in rat.

OBJECTIVES: Application of vitamin K to the skin has been used for suppression of pigmentation and resolution of bruising. However, in rats, no study was reported on its effect regarding wound healing. Thus, the present study was designed to examine the healing effects of creams prepared from vitamin K1 on full-thickness wound in rats. MATERIALS AND METHODS: For inducing full-thickness wound in rats, the excisional wound model was used. Five groups consisting of 8 rats each were used. Vitamin K cream (1% and 2%, w/w) was prepared in eucerin base and applied on the wound once a day until complete healing had occurred. Healing was defined by decreased wound margin (wound contraction), re-epithelialization, tensile strength and hydroxyproline content. Histopathological examination was also done. RESULTS: The effects produced by the topical vitamin K showed significant (P < 0.01) healing when compared with control group in parameters such as wound contraction, epithelialization period, hydroxyproline content and tensile strength. Histopathological studies also showed improvement with vitamin K. CONCLUSIONS: Topical vitamin K demonstrates wound healing potential in full-thickness wound model.

Potentiation of indomethacin-induced anti-inflammatory response by montelukast in formalin-induced inflammation in rats.

The leukotrienes and prostaglandins are biologically active metabolites derived from arachidonic acid. The leukotrienes have a role in inflammatory diseases such as allergic rhinitis, inflammatory bowel disease and asthma. Montelukast, a cysteinyl leukotriene receptor antagonist, is claimed to be effective in asthma. The present study aimed to assess the role of cysteinyl leukotriene receptor antagonist on peripheral inflammation and whether montelukast treatment enhances the anti-inflammatory effect of indomethacin. Anti-inflammatory response was measured using a plethysmometer. Histopathologic examination for leukocyte accumulation was done. Montelukast (0.5-2mg/kg, i.p.) produced a significant anti-inflammatory effect in dose dependent manner against formalin-induced rat paw oedema at 1h but not in 3 and 5 h. When indomethacin (5 mg/kg, i.p) was co-administered with montelukast (1 mg/kg, i.p), the anti-inflammatory effects of indomethacin were significantly increased as compared to the per se effect at 3 and 5 hour after formalin challenge. In histopathology it has been found that combination therapy significantly decreased migration of leucocytes into the site of inflammation. These results show that montelukast has anti-inflammatory properties in peripheral tissue and markedly potentiates the anti-inflammatory activity of indomethacin at 3 and 5 h. It is expected that combination of montelukast with cyclooxygenase inhibitor would prove to be a novel approach to manage complex inflammatory conditions.