Veterinary Research Forum

Abstract The present study evaluated the protective effects of royal jelly (RJ) on methotrexate (MTX)-induced testicular damage in rats, focusing on oxidative stress and autophagy. Methotrexate, a folic acid analogue used in cancer and autoimmune treatments, impairs spermatogenesis via oxidative stress and apoptosis. Twenty-four male Wistar rats were randomized into four groups: Control (normal saline, 35 days), MTX (0.30 mg kg-1, gavage, three times per week, 35 days), MTX + RJ (0.30 mg kg^-1 MTX + 0.10 mg kg^-1 RJ, gavage, three times per week, 35 days), and RJ (0.10 mg kg^-1, gavage, three times per week, 35 days). After 35 days, rats were euthanized and testicular tissue was analyzed via histopathology, immunohistochemistry for LC3-I/II expression in germ cells and qRT-PCR for mRNA expression of autophagy-related genes (Beclin-1, Atg7, LC3-I). Histopathological findings revealed that MTX caused severe interstitial edema, coagulative necrosis and disrupted spermatogenesis with reduced seminiferous tubule diameter, epithelial thickness, tubular differentiation index (TDI) and spermiogenesis index. Co-administration of RJ significantly improved seminiferous tubule morphology, diameter, epithelial thickness, TDI and spermiogenesis index. Immunohistochemistry showed a significant increase in LC3-I/II+ germ cells (spermatogonia, spermatocytes, spermatids) in the MTX group which was markedly reduced in the MTX + RJ group. Similarly, qRT-PCR analysis demonstrated elevated mRNA levels of Beclin-1, Atg7, and LC3-I in the MTX group which were significantly reduced in the MTX + RJ group. These findings suggested that RJ mitigated MTX-induced testicular damage by reducing oxidative stress and autophagy, thereby, preserving spermatogenesis and testicular integrity.

Abstract Crocetin (CRT) is one of the active chemical compounds of saffron and has many biological effects such as antioxidant property. The present study investigated the effects of CRT on crushed sciatic nerve function. Vitamin (Vit) C was used as an antioxidant drug. Thirty rats were divided into six groups including intact, sham, crush, CRT 7.50, CRT 30.00 and Vit C 100. Nine other rats with no surgery were scheduled in three groups to receive 7.50 and 30.00 mg kg^-1 CRT and 100 mg kg^-1 Vit C. In anesthetized rats, right sciatic nerve was crushed using a small hemostatic forceps. Sciatic functional index values on days five, 10, 15 and 20 after crush were accelerated, the severities of sciatic nerve degeneration and gastrocnemius muscle atrophy were ameliorated, and the increased malondialdehyde level and the decreased superoxide dismutase activity in the serum were restored by 20 consecutive days of oral administration of 30.00 mg kg^-1 CRT and 100 mg kg^-1 Vit C. No significant differences were observed between 30.00 mg kg^-1 and 100 mg kg^-1 Vit C. The groups that did not have surgery but received CRT (7.50 and 30.00 mg kg-1) and Vit C (100 mg kg-1) showed no behavioral, histopathological and biochemical alterations when compared to intact group. It was concluded that CRT and Vit C produced similar improving effects on crushed-injured sciatic nerve function. Inhibition of oxidative stress, enhancement of endogenous antioxidant activity might be involved in improving effects of CRT and Vit C.

Abstract This study aimed to evaluate malondialdehyde (MDA) expressions using the immunohisto-chemical method in order to reveal the role of lipid peroxidation in the development and progression of hepatocellular carcinoma (HCC) induced by diethylnitrosamine (DEN) in male Wistar albino rats. Avidin-biotin-peroxidase method was used for immunohistochemical evaluation. Histopathological examinations revealed that DEN caused a mixed pattern (trabecular and acinar formations) of HCC in the majority of rats. The MDA positive stainings were significantly increased in rats in the HCC group compared to the healthy rats in the control group. In conclusion, this study data contain three important findings. The first one is that DEN triggers the formation of reactive oxygen species (ROS), excessively produced ROS cause oxidative stress, and as a result, oxidative stress strongly causes lipid peroxidation. Secondly, it is clear that there is an important relationship between oxidative stress-induced lipid peroxidation and HCC progression. At the same time, MDA is an useful biomarker in determining the prognosis of patients with HCC. The third and final finding is that intra-peritoneal DEN injection once a week for 20 weeks, but not in combination with other promoting chemical agents, appears to be very effective in inducing experimental HCC.

Abstract Diazinon (DZN) is a widely used organophosphate. We studied the effect of quercetin pegylated liposome (QPEGL) on acute low dose DZN-induced oxidative stress and behavioral disorders through monitoring brain serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) in mature male rats. Animals were treated in two control groups that received a single dose of normal saline and dimethyl sulfoxide, and four groups that received a single dose of DZN 10.00 mg kg^-1 (DZN), DZN 10.00 mg kg + quercetin 20.00 mg kg^-1, DZN 10.00 mg kg^-1 + PEGL 20.00 mg kg^-1, DZN 10.00 mg kg^-1 + QPEGL 20.00 mg kg^-1 (QPEGL), respectively. Performances of the rats were investigated by the open field and elevated plus maze tests. Twenty-four hr after the treatments, animals’ brains were harvested and frozen at – 80.00 ˚C. Brain tissues 5-HIAA level was determined by the enzyme-linked immunosorbent assay. Furthermore, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels were determined for oxidative stress analysis. The motor activity was significantly reduced in the DZN group compared to the control group following increased anxiety-like behavior and ameliorated by QPEGL. Moreover, 5-HIAA and MDA levels notably increased in the DZN group compared to the control group and significantly decreased in the QPEGL group compared to the DZN group. The SOD and GPx contents were not significantly changed in the DZN group compared to the control; although, these parameters improved after treatment with QPEGL. Acute low dose DZN exposure resulted in lipid peroxidation and elevated levels of the serotonin metabolite (5-HIAA), leading to neurobehavioral disorders, such as anxiety-like behavior and impaired motor activity, which were alleviated by QPEGL.

Abstract Phthalate esters, such as dibutyl phthalate (DBP), are extensively utilized and human and animal exposure leads to serious toxic effects, including hepatotoxicity. In the present study the protective effects of royal jelly (RJ) on DBP-induced liver damage was investigated. A total number of 40 Wistar albino rats were randomly divided into eight groups (n = 5): control (corn oil), DBP (500 mg kg^-1), RJ (200 mg kg¹), Quercetin (QCN; 50.00 mg kg^-1), RJ (100 mg kg^-1) + DBP, RJ (200 mg kg^-1) + DBP, RJ (300 mg kg^-1) + DBP, QCN (50.00 mg kg^-1) + DBP. After 28 days of daily oral gavage treatment, animals were euthanized. The insulin resistance index, lipid profile and hepatic enzymes were measured on the collected serum samples. Moreover, oxidative and nitrosative stress biomarkers were determined in the liver. Histopathological alterations and ultimately cytochrome P450 2E1 (CYP2E1) activity was also assessed. Data obtained revealed that RJ significantly reduced the insulin resistance index and liver enzymes level in RJ-DBP groups. At the same time, RJ recovered the DBP-induced oxidative stress and restored the DBP-depleted glutathione. Moreover, RJ improved lipid profile and reduced significantly the DBP-induced hepatic CYP 2E1 activity in RJ-DBP groups. Dibutyl phthalate induced-hepatic damage such as necrosis of hepatocytes and scattered bleeding was alleviated in RJ-DBP group. Our data suggested that the administration of RJ could protect the DBP-induced hepatic functional and structural alterations. The RJ protective effects might be attributed to its antioxidant and anti-inflammatory properties and reduced CYP 2E1 activity.

Abstract Cardiac mitochondrial dysfunction is an important feature of aged heart. However, there is still no potent agent to ameliorate cardiac function abnormalities in aged hosts. Olive oil (OLO), containing monounsaturated fatty acids, has diverse protective effects on the cardiovascular system, including anti-diabetic, anti-inflammatory, and anti-hypertensive effects. We evaluated the beneficial impacts of OLO against aging-related cardiac dysfunction. Wistar rats were randomly allotted into three groups with eight rats, including control, aged rats receiving D-galactose (D-GAL), and aged rats administrated with D-galactose plus OLO (D-GAL + OLO). Aged animals were received D-GAL at a dose of 150.00 mg kg^-1 daily through intra-peritoneal injection for aging induction. The animals in D-GAL + OLO group were co-administrated with oral OLO at a dose of 1.00 mL kg^-1 by gavage feeding daily. The administration term was eight weeks. A histological examination of heart tissue was performed. The heart tissues were also harvested to assay the oxidative stress and molecular parameters. The aged animals showed cardiac hypertrophy, increased malondialdehyde level and Bax expression, and reduced mitofusin 2, phosphatase and tensin homologue-induced putative kinase 1, dynamin-related protein 1, and Bcl2 expressions in comparison with the control animals. The OLO treatment ameliorated all these parameters. Overall, OLO could improve cardiac aging through reducing oxidative stress, enhancing genes mediated mitophagy, and improving genes mediated apoptosis in the heart.

Abstract Various companion birds, including budgerigars, are anesthetized with injectable anesthesia. The current study aimed to evaluate oxidative stress indices including malondialdehyde (MDA), total antioxidant capacity (TAC), total oxidant status (TOS), and oxidative stress index (OSI) along with clinical parameters such as the time required to induce, maintain and recover from medetomidine-ketamine anesthesia and midazolam-ketamine anesthesia in budgerigars. Among 20 mature and healthy budgerigars, three groups were assigned as follows: Control (n = 4) to determine baseline oxidative stress indices medetomidine + ketamine (n = 8) anesthetized by intramuscular injections of medetomidine (0.04 mg kg^-1) and ketamine (30.00 mg kg^-1) in the pectoral muscles, midazolam + ketamine (n = 8) anesthetized by intramuscular injections of midazolam (1.00 mg kg^-1) and ketamine (50.00 mg kg^-1). Half of birds (n = 4) in the second and third groups were euthanized by cervical dislocation 1 hr after anesthesia induction, blood samples were collected directly from the heart, and sera were extracted. Additionally, the remaining birds were euthanized 24 hr later, and their serum was analyzed for oxidative stress indices. Clinical parameters were recorded during the study. Compared to the medetomidine + ketamine group, the midazolam + ketamine group experienced shorter induction, anesthetic, and recovery times. Administering medetomidine and ketamine elevated TOS levels compared with midazolam + ketamine. No significant difference was found between the test groups for TAC, MDA, or OSI. Therefore, the midazolam + ketamine regimen appears superior to medetomidine + ketamine when performing minor surgeries on budgerigars.

Abstract Studies conducted on animal models have shown that the administration of glycerol can lead to kidney tissue damage and impaired renal function. This is believed to be caused by oxidative stress and inflammation, which in turn can result in elevated levels of blood urea nitrogen (BUN) and creatinine. These metabolites are commonly used as indicators of renal function. The aim of the current experimental research was to investigate the protective efficacy of ellagic acid in a rat model of rhabdomyolysis induced by glycerol. Sixty healthy adult male Wistar rats weighing between 250 - 300 g were divided into five equal groups including control, rhabdomyolysis (administered 8.00 mL kg^-1 of glycerol), and three rhabdomyolysis plus various doses of ellagic acid (25.00, 50.00 and 100 mg kg^-1 per day; 72 hr after receiving glycerol for 14 days successively) groups. Serum levels of BUN, creatinine, lactate dehydrogenase, alkaline phosphatase, electrolytes and inflammatory cytokines were evaluated in all rats. Histopathological studies were also performed on kidney tissues from all groups. The administration of ellagic acid resulted in a significant increase in renal function biomarkers compared to the rats with acute kidney injury. This increase was consistent with notable reductions in tumor necrosis factor-α levels and increases in interleukin-10 levels observed in blood samples. Furthermore, the improvement in histopathological indices observed in rats received ellagic acid confirmed its nephroprotective role. The results of the current experimental study suggest that ellagic acid can improve kidney damage following glycerol injection, potentially by modulating the inflammatory process.

Abstract About a third of human infertility is related to male factors. Of these, idiopathic-related infertility is not curable. Diabetes mellitus is a metabolic disorder affecting male impotence and fertility by increased production of free radicals and oxidative stress. Saponin, a glycosidic compound found in many plants, improves sperm parameters. The present study investigated the effect of saponin on sperm oxidative stress and testicular structure in streptozotocin (STZ)-induced diabetic mice. The diabetes was induced by the administration of 150 mg kg^-1 STZ via a single intra-peritoneal injection. All experimental mice were allocated to the following groups: Control group, diabetic control group, diabetic group administrated 100 mg kg^-1 saponin daily and one healthy group administrated saponin daily for 56 days. At the end of the treatment period, serum levels of insulin, glucose and oxidative stress markers were measured. A histological evaluation of testicles was performed. Treatment of diabetic mice with saponin ameliorated testicular tissue damage as well as serum glucose and insulin concentrations. Furthermore, in the diabetic group, the serum concentration of malondialdehyde was increased; while, the activity of superoxide dismutase and glutathione peroxidase enzymes was reduced. The mean Johnsen's score and the diameter and thickness of seminiferous tubules were lower in the diabetic mice than control ones. However, these parameters were higher in the saponin-treated mice than controls. Overall, saponin administration rectified all examined parameters. The anti-oxidant role of saponin improves sperm parameters and diabetes-induced testicular oxidative damage.

Abstract Sepsis is an acute condition caused by the systemic inflammatory response syndrome to an infection. There are very few drugs that could improve the severe conditions in patients with sepsis. Hence, it is important to consider different treatment options. In this survey, we studied the effect of adenosine N1-oxide (ANO) and pioglitazone on rats with cecal ligation and perforation (CLP). They were randomly divided to four groups (n = 10) including Group A: as control group receiving normal saline, Group B: the rats with CLP as surgical control group, Group C: the rats receiving ANO, and Group D: the rats receiving pioglitazone. Interleukin (IL) -6, IL-1β, tumor necrosis factor alpha (TNF-α), nitric-oxide (NO) in serum blood and superoxide dismutase (SOD), catalase (CAT) malondialdehyde, (MDA) and myeloperoxidase (MPO) in liver and spleen homogenates were measured. The amount of antioxidant enzymes in the spleen and liver, and finally cell viability and rats’ survival were investigated. The measurement of blood serum nitric-oxide and survival of all groups of rats were also performed. The results indicated that both drugs could cause a decrease in IL-1β, IL-6, TNF-α and NO in rat blood serum and MDA and MPO in the liver and spleen homogenates, however, a significant increase in SOD and CAT in the liver and spleen homogenates in rats that received ANO and pioglitazone was observed compared to rats with CLP group. Cell viability and rats’ survival were significantly improved in rats that received ANO and pioglitazone compared to rats with CLP group. Adenosine N1-oxide showed stronger and more effective effects.

Abstract Ischemia-reperfusion (IR) injury to the lower extremities causes damage to various tissues, notably the limbs. Because research in recent years have demonstrated that saffron and its components are useful in ischemic strokes, the goal of this study was to see whether Crocin (Cr), one of the active constituents in saffron, could protect the gastrocnemius muscle from IR injury. A total number of 32 Sprague-Dawley rats were randomized into four groups randomly: control, Cr, IR, and IR + Cr. Xylazine and ketamine were used to anesthetize all of the rats. The left lower limbs of the other two groups were subjected to 2 hr of ischemia and 2 hr of reperfusion with tourniquet, with the exception of the control and Cr groups. Tumor necrosis factor alfa (TNF-α), interleukin 6 (IL-6), IL-1β, total antioxidant status (TAS) and total oxidant status (TOS) levels were assessed in the blood as well as muscle IL-6, IL1β, SOD1-2, catalase (CAT) and glutathione peroxidase (GPx) expression. According to the IR group, increases in TAS levels and decreases in TNF-α, IL-6, and IL-1β levels were substantial in the Cr therapy group. Cr significantly reduced IL-6 and IL-1β mRNA expression levels in the muscle of the IR group and increased superoxide dismutases 1 (SOD1), SOD2, catalase (CAT), and GPx. Our data showed that Cr protected the gastrocnemius muscle from IR injury in rats and reduced inflammatory markers significantly. These effects of Cr might have been mediated by improved antioxidant enzyme activity, suppression of free radical generation and reduction of oxidative stress.

Abstract Linguatula serrata is a worldwide zoonotic food-borne parasite. The parasite is responsible for linguatulosis and poses a concern to human and animal health in endemic regions. This study investigated the hematological changes, oxidant/antioxidant status and immunological responses in goats and sheep naturally infected with L. serrata. Hematological changes, antioxidant enzymes and malondialdehyde (MDA) levels were measured. The level of inter-leukin-2 (IL-2), IL-4, IL-5, IL-10, and tumor necrosis factor alpha (TNF-α) mRNA expression was investigated in lymph nodes. According to the hemogram results, eosinophils were significantly increased in the infected goats and sheep, and Horizontal Gene Transfer (HGT), hematocrit (HCT), and mean corpuscular hemoglobin concentration (MCHC) were significantly decreased. The levels of MDA and the activity of glutathione peroxidase (GPx) were significantly higher in infected animals than in non-infected animals. However, the activity of superoxide dismutase (SOD) and catalase (CAT) was significantly lower in infected animals than in non-infected animals. A comparison of the cytokine mRNA expression in lymph nodes from infected and non-infected animals showed higher cytokine expression in the infected animals. Infection with L. serrata caused microcytic hypochromic and normocytic hypochromic anemia in goats and sheep. The inconsistent results of immunological changes were found in infected goats and sheep. In both animals, oxidative stress occurred and led to an increase in lipid peroxidation. L. serrata created a cytokine microenvironment biased towards the type 2 immune responses.

Abstract Polycystic ovarian syndrome (PCOS) is a complex endocrine and metabolic disorder. Chlorogenic acid (CGA) bears antioxidant properties with protective effects on different tissues. This study was conducted to evaluate the effect of CGA on follicular development, hormonal status and biomarkers of oxidative stress in a rat model of PCOS. In this experimental study, 18 rats were divided into three equal groups including: control, non-treated PCOS [(estradiol valerate (EV): 40.00 mg kg^-1 intramuscularly)], and PCOS-CGA (EV: 40.00 mg kg^-1 intramuscularly and CGA: 100 mg kg^-1 intraperitoneally once a week for eight consecutive weeks). At the end of treatment period, all rats were anesthetized. Then 5.00 mL blood samples of rats in the three groups were taken and prepared for hormonal analyses and their ovaries were isolated and dissected mechanically free of fat and mesentery. The ovaries underwent the following analyses: Morphological study with Hematoxylin and Eosin staining and biochemical study using the malondialdehyde (MDA) level and total antioxidant activity. Data were analyzed using one-way ANOVA and post hoc Tukey’s test. The serum level of luteinizing hormone, estrogen, testosterone, antioxidant capacity, glutathione and the number of cystic follicles in the PCOS group treated with 100 mg kg^-1 Chlorogenic acid compared to the non-treated PCOS group were significantly decreased, however, the serum level of follicle stimulating hormone, progesterone, MDA and the number of secondary, graafian follicles and corpus luteum were significantly increased. Chlorogenic acid could be effective in ameliorating follicular development as well as hormonal and biochemical disorders in rats with PCOS.

Abstract The anti-diabetic effects of Ribes khorasanicum as a traditional remedy were investigated in diabetic rats. Thirty-five male rats were divided into five groups: control, diabetic, diabetic treated with metformin (300 mg kg^-1; D+Met), diabetic treated with 250 and 500 mg kg^-1 of Ribes khorasanicum hydro-ethanolic extract (D+Rib250 and D+Rib500). After six weeks of treatment, sera of overnight fasted animals were collected and used for measurement of glucose, insulin, lipid profile, urea, creatinine, and hepatic enzymes levels. Moreover, liver and kidney of rats were removed and used for measurement of oxidative stress including malondialdehyde (MDA), thiol content, and the activity of catalase (CAT) and superoxide dismutase (SOD). Streptozotocin (STZ)-induced diabetes increased the levels of serum glucose, triglycerides (TG), total cholesterol (TC), and LDL-C, urea, creatinine, hepatic enzymes, and kidney and liver oxidative stress markers, while decreased insulin and HDL-C when compared to control group. In all treated groups serum levels of glucose, TC, LDL-C, TG, and urea were decreased, while liver SOD activity was increased compared to the diabetic group. The D+Rib500 group had lower Serum glutamic pyruvic transaminase (SGPT), creatinine, and kidney MDA levels, but higher insulin, HDL-C levels, liver CAT activity, and kidney thiol content, and CAT activity compared to diabetic group. In D+Met group, serum levels of serum glutamic-oxaloacetic transaminase (SGOT), creatinine, and MDA of liver and kidney were decreased, while liver SOD activity was increased compared to the diabetic group. Based on our findings, treatment with Ribes khorasanicum improved diabetic complications, while the effect of a higher dose of the extract was comparable to metformin’s.

Abstract The objective of the present study was to evaluate the alterations in selected indicators of immune responses and oxidative stress of broilers fed with nano-manganese. One hundred-sixty 1-day-old broiler chicks were randomly assigned into four groups with three replicates. Birds were fed the same basal diet supplemented with nano-manganese oxide, as 0.00 (control group), 50.00, 100, or 150 mg kg^-1 of diet. The birds were vaccinated against avian influenza (AI), Newcastle disease (ND), infectious bronchitis (IB) and infectious bursal disease (IBD) as the standard vaccination schedule. Blood sample was taken from the brachial vein of birds on 42^th day. A significant decrease in antibody titer against sheep RBC was revealed in the nano-manganese 100 and 150 groups compared to the control group. In addition, the antibody titers against IB and ND were significantly lower in the all nano-manganese groups compared to the control group. No significant difference was observed for the antibody titer against AI and oxidative stress indices among the experimental groups. The findings in the present study suggested that nano-manganese at 50.00, 100 and 150 mg kg^-1 levels might suppress humoral immune response in broilers which should be taken into consideration in supplementation.

Abstract The present study was conducted to evaluate the effects of different doses of haloperidol (HP) on induction of oxidative stress in blood and liver cell degeneration in comparison with influences of HP pre-treatment on inflammatory process induced by intraperitoneal (IP) administration of lipopolysaccharide (LPS). One hundred twenty male albino Wistar rats were randomly divided into eight groups (15 in each), including: Control group, LPS group, three groups as HP administration in three divided doses (0.50, 1.00 and 2.00 mg kg^-1), and three treatment groups that HP was administered in three doses (0.50, 1.00 and 2.00 mg kg^-1) prior to LPS administration.Concentrations of malondialdehyde, activities of antioxidant enzymes including glutathione peroxidase, superoxide dismutase and also the levels of tumor necrosis factor-alpha and interleukin 1-beta were measured in blood and serum. In addition to liver histopathological changes evaluation, hepatic silent information regulator of transcription 1 (SIRT1) and phosphorylated-nuclear factor-κB (p-NF-κB) levels were quantitated. Our findings indicated that sole administration of HP (particularly higher doses) can induce oxidative stress in blood and cell degeneration in liver, while it can attenuate inflammatory process induced by LPS administration presumably via SIRT1 up-regulation and preventing the induction of p-NF-κB. The oxidative and degenerative effects of HP and its impact on inflammatory status were completely dose- dependent according to our results. The possible anti-inflammatory effects of HP may affect reparative mechanisms and hepatic cell degeneration. However, the influences of HP on immune system need further investigations and its higher doses should be administered cautiously especially in patients with immune system dysfunctions.

Abstract Increasing applications of carbon nanotubes (CNTs) indicate the necessity to examine their toxicity. According to previous studies, CNTs caused oxidative stress that impaired β-cell functions and reduced insulin secretion. Our previous study indicated that single-walled carbon nanotubes (SWCNTs) could induce oxidative stress in pancreatic islets. However, there is no study on the effects of multi-walled carbon nanotubes (MWCNTs) on islets and β-cells. Therefore, the present study aims to evaluate effects of MWCNTs on the oxidative stress of islets and the protective effects of caffeic acid (CA) as an antioxidant. The effects of MWCNTs and CA on islets were investigated using MTT assay, reactive oxygen species (ROS), malondialdehyde (MDA), activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), the content of glutathione (GSH) and mitochondrial membrane potential (MMP) and insulin secretion measurements. The lower viability of islet cells was dose-dependent due to the exposure to MWCNTs according to the MTT assay. Further studies revealed that MWCNTs decreased insulin secretion and MMP, induced ROS creation, increased the MDA level, and decreased activities of SOD, GSH-Px, CAT, and content of GSH. Furthermore, the pretreatment of islets with CA returned the changes. These findings indicated that MWCNTs might induce the oxidative stress of pancreatic islets occurring diabetes and protective CA effects that were mediated by the augmentation of the antioxidant defense system of islets. Our research suggested the necessity of conducting further studies on effects of MWCNTs and CA on the diabetes.

Abstract Acetamiprid (ACP) belonging to the neonicotinoid family used against wide array of pests in agriculture and domestic purposes. In this study, we evaluated the attenuating effects of ethanolic extract of Mangifera indica leaves (EEMI) in averting reproductive toxicity caused by ACP in male guinea pigs. Thirty male guinea pigs were randomly assigned to five treatment groups (n = 6). Group 1 (T0) received distilled water orally; group 2 (T0-) was given 80 mg kg^-1 of ACP and groups 3, 4 and 5 were treated, respectively, with EEMI at doses of 50, 100 and 200 mg kg^-1 plus ACP. After 90 days, the reaction time, sexual organ weights, sperm count, motility and anomalies, spermatozoa with entire plasma membrane, testicular histology, serum testosterone concentration, testicular malondialdehyde (MDA) level, reduced glutathione (GSH) concentration, testicular superoxide dismutase (SOD) and catalase (CAT) activities were assessed. Co-administration of EEMI significantly reduced the reaction time, sperm anomalies and testicular MDA, SOD and CAT levels compared to the T0- group. Co-treatment of EEMI significantly alleviated sperm count and motility, percentage of spermatozoa with the normal plasma membrane, serum testosterone concentration, accessory sex gland weights, and testicular GSH concentrations. The ACP treatment induced cell membrane degradation in the testis and this effect was prevented with the addition of EEMI. In conclusion, ACP negatively affected the animal reproductive function and induced oxidative stress. The addition of EEMI alleviated the toxic effects of ACP on the reproductive function of male guinea pigs.

Abstract The aim of this study was to evaluate the potential use of a DNA comet assay, DNA fragmentation fluorimetric assay and reactive oxygen species levels as potential biomarkers of genome conditions of dental pulp stem cells (DPSCs) isolated from dog canine teeth. Mesenchymal stem cells were isolated from the dental pulp collected from dog teeth. The results obtained suggest the ideal moment for clinical application of cellular therapy for this type of cell. The cell culture was maintained with Dulbecco’s modified Eagle’s medium supplemented with 10.00% fetal bovine serum for eight passages. During each passage, cell proliferation, oxidative stress and level of DNA fragmentation were assessed by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay, testing 2,7 dichlorodihydro-fluorescein-diacetate and PicoGreen^®, respectively. There were important differences among the first three DPSC passages compared to passages 4–8 and a large number of nuclei with some levels of DNA damage (30.00 to 40.00% in initial DPSC passages and > 50.00% in late passages), indicating in vitro DPSC genomic fragility. Within the limitations of this study, the results suggest these relatively simple and inexpensive approaches - comet and DNA fragmentation assays - could help sort stem cells with less DNA damage for use in research or therapies.

Abstract Cyclophosphamide is a chemotherapy drug for the treatment of cancer. Chicken embryo amniotic fluid, vitamin C and coenzyme Q10 have anti-oxidant properties. Total of 70 adult female mice were selected and divided into seven groups. The first group that received 2 ml kg^-1 of inactivated amniotic fluid subcutaneously. The second group treated with 75 mg kg^-1of cyclophosphamide by intraperitoneal injection. Third to fifth groups received 1, 2, and 4 ml kg^-1 of chicken embryo amniotic fluid, respectively. The sixth group received vitamin C at a dose of 0.2 mg g^-1 of body weight by oral gavages. Seventh group received 10 mg kg^-1 coenzyme Q10 intraperitoneally. All cyclophosphamide treated groups (3-7) received 75 mg kg^-1 of cyclophosphamide intraperitoneal on day 22. The mice were euthanized on day 29 and ovarian tissue antioxidant enzymes including glutathione peroxidase, superoxide dismutase and catalase activities and malondialdehyde (MDA) were evaluated. Activities of above mentioned enzymes in treatment groups (3-7) was significantly higher than patient control group (2). The results also revealed that MDA levels were higher in the control group in comparison to other treatment groups. Therefore, it is concluded that the chick embryo amniotic fluid and coenzyme Q10 can compete with compounds like vitamin C in increasing the anti-oxidant level in ovarian tissue.

Abstract Bisphenol-S (BPS) is a new bisphenol-A substitute widely used in many plastic products. Bisphenol-A as a main member of bisphenol family has been known as an endocrine system disrupter chemical compound. Like other members of bisphenol family, there is public health concern about the toxic effects of BPS on reproductive system, thus, we examined BPS effects on in vitro fertilization (IVF) potential and oxidative stress status in a murine model. Adult female mice (n = 70) were randomly divided into control and BPS-treated groups. Bisphenol-S was administered at doses of 0, 1, 5, 10, 50 and 100 µg kg^-1 body weight per day intraperitoneally for 21 consecutive days. Twenty-Four hr after the last treatment, five mice in each group were super-ovulated and the oocytes were harvested for IVF. All ovaries were collected and used for biochemical factors analyses. Bisphenol-S exposure at doses more than 10 µg kg^-1 induced developmental arrest of pre-implantation embryos. Further, lipid peroxidation measurement in ovaries indicated that all doses of BPS cause oxidative stress in female mice. In conclusion, BPS administration even in low doses can result in female reproductive toxicities and oxidative stress in mice.

Abstract Nicotine (NIC) adversely influences male reproductive system. Achillea millefolium (Achm) as a medicinal plant is highly regarded for its antioxidant and anti-inflammatory properties. The present study was conducted to assess whether Achminflorescences alcoholic extract could serve as a protective agent against reproductive toxicity in NIC-exposed male rats. Adult male rats were randomly divided into six groups. Two groups received NIC at doses of 0.20 and 0.40 mg kg^-1 per day in 0.50 mL sterile distilled water for 48 days intraperitoneally, respectively. The further two groups received NIC at doses of 0.20 and 0.40 mg kg^-1 per day in 0.50 mL sterile distilled water for intraperitoneally along with Achm extract at a dose of 1.20 g kg^-1 per day in 1 mL sterile distilled water orally for 48 days, respectively. A vehicle treated control group and an Achm-only treated group were also included. The NIC-exposed groups showed significant reductions in epididymal sperm count, motility, viability and serum levels of FSH, LH and testosterone as well as testicular antioxidant capacity. Moreover, the incidence of apoptosis and abnormality in spermatozoa along with testicular malondialdehyde and total nitrite levels were significantly higher in NIC-treated rats. The above-mentioned parameters were restored to near normal levels by Achm co-administration. These findings indicated thatAchmmay partially be protective against NIC-induced testicular toxicity.

Abstract Mycophenolate mofetil (MMF) is a selective inhibitor of Inosine-5′-monophosphate dehydrogenase. Gastrointestinal (GI) disturbances in immature ones are reported for MMF-induced compilations, which in the case of occurrence dose reduction is required. Thus, in the present study, the fructooligosaccharide raftilose^® (RFT) was co-administrated with MMF to estimate the protective effect of RFT against MMF-induced GI complications. Thirty six immature male Wistar rats were divided into six groups including: Control (normal saline), RFT-treated (100 mg kg^-1), MMF-treated (20 mg kg^-1), MMF + LRFT (50 mg kg^-1), MMF + MRFT (100 mg kg^-1) and MMF + HRFT (200 mg kg^-1) groups. The hematocrit (Hct), lymphocyte/total WBC, feces water content and pH were analyzed. Moreover, the hepatic functional tests, kidney-related biomarkers, lipid and protein profiles, total antioxidant capacity (TAC), malondialdehyde (MDA) and nitric oxide (NO) contents were assessed. Co-administration of RFT stabilized the MMF-reduced body weight. The MMF significantly diminished Hct and lymph/total WBC (p < 0.05). Only MRFT enhanced the lymphocyte/total WBC. Increased water content, no changes in feces pH, increased serum ALT and AST, no alteration in urea and mild enhancement in creatinine were demonstrated in MMF-received animals. However, RFT at low dose ameliorated the feces parameters and reduced ALT. No significant changes were demonstrated for serum lipid and protein profiles in MMF- and RFT + MMF-treated groups. The RFT enhanced the serum TAC, reduced MDA and NO contents. In conclusion, our data suggested that RFT could be considered as an effective agent to subsidize the MMF-induced clinical, hematological and biochemical disorders.

Abstract Grape seed proanthocyanidin (GSP) bears a very powerful antioxidant effects. Studies demonstrated that proanthocyanidins protect against free radicals mediated cardiovascular and renal disorders. The present study was designed to assess the effect of GSP on the heart of diabetic rats. Forty rats were divided into four groups of 10 animals each: Group I: control, Group II: control group were given GSP, Group III: diabetic group, Group IV: diabetic group treated with GSP. Diabetes was induced by a single dose of streptozotocin, and then GSP (200 mg kg^-1 body weight) was administrated for four weeks. Blood glucose, glycosylated hemoglobin (HbA1c) and also the levels of lipid peroxidation and antioxidant enzymes were examined in the heart tissues of all groups. Oral administration of GSP to diabetic rats significantly reduced (p < 0.05) heart weight, blood glucose, HbA1c and lipid peroxidation level, but increased (p < 0.05) body weight and activities antioxidant enzymes when compared to diabetic group. The results indicated that GSP could be useful for prevention or early treatment of cardiac disorder caused by diabetes.

Abstract Toxic effects of monensin, a polyether antibiotic mainly used as a coccidiostat, have been described in a wide range of animals. The present study was performed to investigate the toxic effects of monensin in goats. Seven adult goats were administered sodium monensin, 13.5 mg kg-1, daily for five consecutive days via gastric gavage. Monensin toxicity was evaluated by clinical signs, serum biochemistry and pathology. Monensin exposure caused diarrhea, tachycardia and reduction in ruminal movements and body temperature. Significant increase of creatine kinase, lactate dehydrogenase, aspartate aminotransferase, total bilirubin, blood urea nitrogen, creatinine and erythrocyte superoxide dismutase were observed in monensin exposed goats. Reduction of erythrocyte glutathione peroxidase and elevation of serum malondialdehyde and troponin I were inconsistent. In necropsy, there were effusions in body cavities, vacuolar degeneration and coagulative necrosis in cardiac and skeletal muscles and renal tubular necrosis. These findings suggested that monensin intoxication in goats leads to cardiac, skeletal and renal damage and a wide range of biochemical abnormalities. Oxidative stress may be involved in the pathogenesis of monensin poisoning.