Veterinary Research Forum

Abstract This study evaluated the protective effects of intraperitoneal tannic acid (TA) against ischemia-reperfusion (I/R) injury in a rat model of testicular torsion. Eighteen adult male Wistar rats were randomized into three groups (n=6 each): sham (surgery without ischemia), I/R (3 hr ischemia + 3 hr reperfusion), and I/R+TA (TA 50 mg kg-1; 100 µL i.p. 30 min before reperfusion). Testicular tissues were sampled immediately after reperfusion for biochemical assays to measure malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels. Epididymides were analyzed 60 days later for sperm count, motility, viability, morphology, and DNA integrity. I/R significantly elevated MDA and sperm DNA damage while reducing SOD, GPx, sperm parameters, testicular weight and spermatogenesis. TA administration reversed these changes and restored the parameters to levels close to those of the sham group. Overall, intraperitoneal TA mitigated I/R-induced oxidative stress and preserved reproductive function, indicating its potential therapeutic value in testicular torsion.

Abstract Testicular ischemia-reperfusion (I/R) injury during testicular torsion is strongly influenced by oxidative stress caused by excessive accumulation of uncaptured reactive oxygen species (ROS). Kisspeptin-10, a biologically active fragment of the kisspeptin peptide family, has demonstrated significant antioxidant and anti-apoptotic properties. Recent studies indicate that kisspeptin-10 can mitigate oxidative stress by reducing reactive oxygen species levels and enhancing the activity of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and catalase. This study examined the effects of kisspeptin-10 on I/R injury in testicular torsion/detorsion (T/D) of male rats. Twenty male rats were divided into four groups: the control group and three treatment groups (720° T/D, 720° T/D+ 0.50 µg kg-1 kisspeptin-10, 720° T/D+ 1.00 µg kg-1 kisspeptin-10). After inducing 720° clockwise testicular torsion for 2 hr, various factors such as sperm parameters, number, total motility, viability, DNA damage and hypoosmotic test were evaluated. The results showed that 720° T/D can increase sperm DNA damage. In addition, it also had negative effects on overall motility and other properties such as viability and plasma membrane functionality. The results also showed that administration of kisspeptin-10 to T/D rats can reduce DNA damage. These effects could also increase overall motility, viability and plasma membrane functionality compared to the T/D group. Based on our results, kisspeptin-10 provides significant protection against acute T/D injury to the testis when administered after spermatic cord torsion in rat.

Abstract Testicular ischemia-reperfusion is accompanied by elevated production of reactive oxygen species. It has been reported that reactive oxygen species are highly reactive to cellular carbohydrates, DNA, lipids and proteins, and result in testicular ischemia-reperfusion injury. Gastrodin is the principal active ingredient isolated from the medicinal plant Gastrodia elata Blume and has anti-oxidative stress effect. In this study, we explored the potential protective activity of gastrodin in rat testicular ischemia-reperfusion injury model and underlying mechanism. Male rats were randomized into 3 groups (sham control, testicular ischemia-reperfusion injury, testicular ischemia-reperfusion injury along with gastrodin injection) (n = 20). Testicular ischemia-reperfusion injury group received 2-hour period of left testicular torsion (720°, counterclockwise) and 4-hour or 3-month period of testicular detorsion. At the onset of testicular detorsion, gastrodin-treated rats were given 100.00 mg kg-1 gastrodin by intraperitoneal route. Following testicular detorsion, testicular tissues were collected for enzymatic activity analysis, oxidative stress evaluation and histopathological examination. The ipsilateral testicular xanthine oxidase activity (source of reactive oxygen species production) and malondialdehyde level (a precise biomarker of reactive oxygen species) were significantly increased in testicular ischemia-reperfusion injury group versus sham control group, while testicular spermatogenic function was decreased. Furthermore, gastrodin administration reduced xanthine oxidase activity and malondialdehyde level in ipsilateral testicular tissue, while improving testicular spermatogenic function. Consequently, it is suggested that gastrodin plays a protective role in testicular torsion/detorsion-induced ischemia/reperfusion injury through inhibiting xanthine oxidase activity to decrease reactive oxygen species formation.

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 Cadmium (Cd) is a highly toxic environmental pollutant known to cause severe damage to the male reproductive system. This study aimed to investigate the protective effects of bee bread (BB), a natural product with anti-oxidant, anti-apoptotic, and anti-inflammatory properties, against Cd-induced testicular toxicity in male Wistar rats. A total number of 32 rats were divided into four groups, including control, BB (0.50 g kg^-1), Cd (5.00 mg kg^-1), and Cd + BB (5.00 mg kg^-1 and 0.50 g kg^-1, respectively) groups. Administrations via oral gavage were performed for 4 weeks. Semen analysis revealed significant reductions in sperm motility and density along with increases in abnormal and dead sperm ratios in the Cd and Cd + BB groups compared to controls. Histopathological examination showed severe degeneration and desquamation of germ cells, tubular atrophy, and a decrease in spermatozoa in the Cd-treated groups. Polymerase chain reaction analysis indicated up-regulation of apoptotic markers (caspase-3, -8, and -9) and oxidative stress enzymes (catalase and superoxide dismutase) in the Cd group, signifying disrupted testicular function. The BB administration partially mitigated Cd-induced damage as evidenced by less severe histopathological changes and moderated gene expression alterations. However, the protective effects of BB were not sufficient to completely counteract the toxic impact of Cd. The present study concluded that while BB had potential in reducing Cd-induced testicular toxicity, its protective efficacy was limited, warranting further research to explore its therapeutic potential in combination with other protective agents.

Abstract In recent years, liver transplantation has emerged as the standard therapy for several liver disorders. Throughout the procedure, the transplanted liver tissue is subjected to varying degrees of ischemia-reperfusion (IR) damage. Consequently, there has been a long-standing pursuit of substances that can alleviate the harm caused by IR. In our investigation, we employed dapagliflozin as a potential therapeutic agent. Eighteen Wistar rats were divided into three groups (n = 6), including treatment, IR, and control that did not undergo surgical intervention. Two days prior to surgery, the treatment group received dapagliflozin at a dosage of 10.00 mg kg^-1 orally. During surgery, liver ischemia was induced for 1 hr, followed by a 24-hr reperfusion period. The IR group exhibited elevated levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, bilirubin, lactate dehydrogenase, and malondialdehyde compared to the control group. In contrast, the treatment group showed levels of these factors that were closer to those of the control group. While total protein, albumin, and total anti-oxidant capacity decreased in the IR group, this decline was less significant in the treatment group. Analysis of oxidative stress in liver tissue revealed that the treatment group had increased anti-oxidant capacity, and exhibited less oxidative stress compared to the IR group. Furthermore, dapagliflozin was found to reduce the degree of liver edema, necrosis, and vascular hyperemia following IR. Overall, dapagliflozin demonstrates the potential to lessen liver damage, enhance liver tissue regeneration, and mitigate the consequences associated with liver impairment.

Abstract Sterilization in animals serves multiple purposes, such as behavior control, performance improvement, and population management. Chemical sterilization has emerged as a promising non-surgical alternative to traditional methods. This study aimed to investigate the effects of intra-testicular injection of zinc-doped carbon dots (Zn-CDs) nanoparticles as a chemical sterilant in mature rats. Twenty-five rats were randomly divided into five groups, including a control group without injection, a sham group receiving 0.50 mL distilled water, and three treatment groups administered respectively 0.50, 2.00, and 8.00 mg kg^-1 of Zn-CDs synthesized through a hydrothermal process. Following anesthesia with ketamine and xylazine, and aseptic preparation, intra-testicular injections were administered bilaterally. At 60th day post-injections, blood samples were collected to measure serum testosterone levels using chemiluminescence immunoassay. The rats were then surgically castrated to assess sperm parameters and testicular histopathology. Testicular oxidant/anti-oxidant status was also evaluated. The results revealed a dose-dependent reduction in sperm viability, membrane integrity, and motility, accompanied by increased sperm DNA damage. The highest Zn-CDs dose caused the most significant decrease in sperm concentration, as well as severe testicular tissue damage. In addition, anti-oxidant capacity, seminiferous tubules maturation, testosterone production, and spermatogenesis declined with increasing Zn-CDs concentrations in a dose-dependent manner. These findings indicate that intra-testicular injection of Zn-CDs effectively induces infertility in mature rats and holds potential as a chemical sterilization method. With further studies to evaluate safety and efficacy, this approach could be developed as a practical solution for large-scale in situ castration, offering a non-surgical alternative for over-population control programs.

Abstract The aim of this study was to investigate the effects of N-butyl-2-cyanoacrylate (NBC), a synthetic tissue adhesive and graft material, on the healing of experimentally induced joint defects in rats. Twenty healthy male Wistar rats with an average body weight of 250 - 300g were used in this experiment. In the experimental group, NBC was given into the induced defects (2.00 mm) and then, they were closed routinely. In the control group, no intervention was made on the defects and they were closed routinely. All animals were observed during 30 days. In the radiological evaluation, the significant differences were recorded from days 15 to 30 between groups. In the histopathological evaluation, the distributions of collagen II and transforming growth factor-beta immuno-positive cells were more intense in experimental group than control group. As a result, the radiological and histopathological evaluations indicated that using NBC in joint defect increased tissue compatibility of newly formed cartilage cells and collagen fibers. However, how NBC provides chondrocyte production and harmony between the damaged and surrounding tissues remains a subject that must be studied.

Abstract The administration of dispersed allogeneic biomaterial (AB) into the para-articular region and joint cavity allows slowing down the processes of tissue destruction in arthritis. The aim of the study was to examine the effect of AB on the course of experimental adjuvant arthritis (AA) in rats. For modeling of AA, complete Freund's adjuvant was injected into the plantar surface of the hind paw of 60 white outbred female rats. The study included intact group, control group, and experimental group. After 37 days of the experiment, blood was collected for hematological analysis and the knee joint with surrounding tissues was harvested for standard histological examination. Intra-articular administration of AB to experimental rats while using complete Freund's adjuvant neutralized the manifestation of signs of a generalized inflammatory process in the joints and reduced the degree of destructive changes in the articular apparatus, preserving the structure of the cartilaginous layer. The use of AB made it possible to stabilize the red and white blood cells levels in the experimental group, as well as significantly increase the reduced level of monocytes. Intra-articular administration of AB during AA modeling exhibits an osteo- and chondro-protective effect, providing positive anti-inflammatory and symptom-modifying effects and weakening the manifestation of pathomorphological changes in the joints of experimental rats.

Abstract Cisplatin (CS) is a broad-spectrum chemotherapeutic agent that causes serious adverse effects, such as cardiotoxicity, despite its potent anti-tumor efficacy. This study aimed to evaluate the cardioprotective effects of betaine in rats exposed to repeated low-dose CS administration using histopathological and immunohistochemical methods. Forty female Wistar albino rats were divided into four groups, including control, betaine, CS, and CS + betaine. Betaine (250 mg kg^-1) was administered orally on a daily basis for four weeks, while CS (8.00 mg kg^-1) was administered intraperitoneally once a week for the same duration. Cardiomyocytes were then examined using histopathological and immunohistochemical methods. The data were analyzed using one-way analysis of variance and Tukey tests. Histopathological analysis revealed cardiomyocyte disorganization, myofibril loss, and increased eosinophilia in the CS group. Betaine treatment partially prevented CS-induced histological damage, contributing to the cardiac muscle structure preservation. Immunohistochemical analyses demonstrated a significant increase in transforming growth factor-beta and interferon gamma expressions in the CS group, whereas betaine administration reduced transforming growth factor-beta levels. Interleukin 6 expression was lower in the CS + betaine group compared to the CS group. No significant differences were observed between groups regarding Interleukin -1β expression. These findings suggest that betaine may have protective effects against CS-induced cardiotoxicity. Its anti-inflammatory properties appear to mitigate cardiomyocyte damage.

Abstract Spinal cord injury (SCI) results in the demise of neural and glial cells, as well as extensive neuro- inflammation. Hydrogel formulation for prolonged release of melatonin (Mel) has demonstrated enhanced effectiveness and safety. In this study, SCI was induced in rats by contusion at the T₉ vertebrae. Chitosan (CH) /Mel hydrogel was fabricated and characterized using scanning electron microscopy (SEM) and Fourier transform infra-red to examine its specific effects on the apoptotic and histopathological markers of SCI. The scanning electron microscopy images revealed the presence of porosity in the CH/Mel hydrogel. Forty male Wistar rats were randomly divided into five groups (n = 8), including sham, control (SCI-induced treated locally with 100 µL CH hydrogel), and groups 3, 4, and 5 (treated locally immediately after SCI induction with 100 µL CH hydrogel containing 50.00, 100, and 200 mg kg^-1 Mel, respectively). The CH/Mel hydrogel at a dose of 25.00 mg mL^-1 significantly increased cell viability in the U87 cell line after 24 hr of exposure. However, at 48 and 72 hr after exposure, Bax and Bcl2 expressions were significantly increased and reduced in the SCI group, respectively, and CH/Mel hydrogel could alleviate their expressions, especially in higher doses. In addition, S100 protein expression was up-regulated in the SCI group. However, CH/Mel hydrogel down-regulated it in a dose-dependent manner. The histopathological findings demonstrated that CH/Mel hydrogel dramatically improved SCI outcomes, like vacuolar degeneration, necrosis, and severe cystic and axonal degenerations. In conclusion, CH/Mel hydrogel induced neuroprotection and it had the potential to be used as a therapeutic agent for the treatment of SCI.

Abstract This study investigated carbendazim (CBZ)-induced hepatic dysfunction and the mechanistic pathway regarding the protective effect of melatonin (MEL). Twenty-eight male rats were grouped as follows: Control, CBZ (150 mg kg^-1), MEL (20.00 mg kg^-1), and CBZ + MEL. The experiment was conducted for 60 days. Tissue samples were stained with Hematoxylin and Eosin and immuno-fluorescence methods to examine apoptotic pathway. Also, hepatic enzymes and miR-122 expression were evaluated. The findings indicated that the CBZ group exhibited an increase in degenerated hepatocytes, hyperemia of sinusoids, and leukocyte infiltration, accompanied by elevated levels of aspartate aminotransferase and alanine aminotransferase, as well as up-regulation of miR-122. Also, there was a significant increase in the fluorescence intensities of caspase-3 and Bax in the CBZ group, whereas a substantial reduction in the fluorescence intensity of Bcl-2 was recorded. In contrast, the simultaneous administration of MEL alongside CBZ was shown to be effective in improving histological structure, decreasing levels of aspartate aminotransferase and alanine aminotransferase, reducing the apoptosis index, and modulating the expression of miR-122 in comparison with the CBZ-only group. The increased expression of miR-122 noted in the CBZ group may correlate with an elevation in the immunoreactivity of apoptosis markers and alterations in liver architecture. Additionally, MEL seems to alleviate CBZ-induced hepatotoxicity by down-regulating miR-122 expression, diminishing the fluorescence intensity of caspase-3 and Bax, and enhancing the immunoreactivity of Bcl-2. Collectively, the regulation of miR-122 may serve as a potential mechanism by which MEL confers its protective effects against liver damage induced by CBZ.

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 Exposure to extreme temperature conditions such as occurs in certain occupations is known to induce male infertility. In humans and most of the mammals, it has been shown that whole-body heat stress (HS) decreases fertility and produces defective embryos. Hence, the present study aimed at gaining some insights into the mechanisms producing defects after HS. In the present study, 56 mature male Wistar rats were randomly categorized into eight groups (n = 7), including group 1: Control, groups 2: Bioactive peptides (BPs; 10.00 mg kg^-1), groups 3, 4, and 5: Heat-stressed (37.00, 39.00, and 43.00 ˚C for 20 min, respectively), and groups 6, 7, and 8: Heat-stressed along with BPs (10.00 mg kg^-1), respectively. All treatments were administered orally once per day. The HS was induced through the immersion of rat scrotums in a water bath. After 45 days, rats were sacrificed and left testes were removed, fixed, and used for histological and immunohistochemical studies. Harvested right testes were also used for oxidative stress assessments and molecular analyses. Heat stress increased testicular tissue damage, elevated oxidative stress and reactive oxygen species production, and increased germ cells apoptosis, p53 and caspase 3 expressions, and Bax/Bcl-2 ratio. Treatment with BPs as a substance with anti-oxidant properties ameliorated the damage caused by HS. The results of this study highlight the protective role of BPs in the reproductive tract under HS. Bioactive peptides may have potential function against testicular tissue oxidative stress and apoptosis.

Abstract Neuroprotective effects for natural products are supported by several studies. In this regard, safranal, a constitute of saffron, has the potential to exert beneficial effects in neurological disorders such as Parkinson's disease, epilepsy, stroke, multiple sclerosis and Alzheimer's disease. Here, we investigated the effect of safranal on penicillin-induced epileptiform activity. Also, the effects of intracerebroventricular (ICV) microinjection of AM251 as a CB1-cannabinoid receptors antagonist to clarify the possible mechanism of safranal were evaluated. Epileptiform activity was induced by intra-cortical administration of penicillin (300 IU, 1.50 μL) in urethane-anesthetized rats. Electrocorticographic recordings were used to analyze the frequency and amplitude of spike waves. Intraperitoneal injections of safranal at doses of 1.00 and 4.00 mg kg^-1 significantly reduced both the number and amplitude of spike waves. The ICV microinjection of AM251 (0.50 μg 2.00 μL^-1) significantly increased the frequency and amplitude of spike waves. In addition, the anti-epileptic effect induced by administration of safranal at a dose of 4.00 mg kg^-1 was partially prevented by ICV microinjection of 0.50 μg 2.00 μL^-1 of AM251. The results showed anti-epileptiform activities for safranal. Central CB1 cannabinergic receptors might be involved in the anti-epileptiform activity of safranal.

Abstract The torsion model of testis in a rat was adopted for evaluation of possible effects of propolis (Prop) on ischemia-reperfusion (IS/REP) injury. The healthy male Wistar rats (totally 24 animals) were randomized into four groups (n = 6) and animals experienced bilateral testicular torsions as follows: In sham group just, laparotomy was performed and in IS group, animals experienced a 3 hr period testicular IS. In IS/REP group, a 3 hr period of IS followed by a 3 hr period of testicular REP for left testis and a one-week testicular REP for right testis were done. In this group animals were gavaged by 1.00 mL normal saline 1 hr before the onset of IS. In IS/REP/ Prop group, the same procedures for IS/REP animals were followed as well as gavage of 1.00 mL Prop extract solution 1 hr before the onset of IS. Analyses of biochemistry, histology, inflammatory biomarkers and sperm parameters were carried out. In IS/REP/Prop group, nitric oxide synthase malondialdehyde, myeloperoxidase and 8-hydroxy-2 deoxyguanine in IS/REP/Prop group were significantly decreased and, superoxide dismutase, total glutathione, glutathione peroxidase, glutathione reductase and glutathione S-transferase were significantly increased compared to the other animals. In IS/REP/Prop group, seminiferous tubules (with normal spermatogenesis) showed all stages of spermatogenic cells with plentiful spermatozoa. Tubular deterioration and atrophy and spermatogenic cell loss in were seen in a limited extent. The mean concentrations of Interleukin-1 beta and tumor necrosis factor alpha in IS/REP/Prop were significantly decreased. Sperm quality was significantly improved by Prop in IS/REP/Prop group. It was concluded that Prop could be supportive in diminishing IS/REP injury in testicular tissue exposed to ischemia.

Abstract This study aimed to investigate the effects of a high-fat and cholesterol diet (HFCD) on rats gastric mucosa. In the study, a total of 16 (40-day-old Sprague Dawley) male rats were used and randomly divided into two groups (each consisted of eight rats). Rats in the control group had no implementations other than normal feeding. For 10 weeks, rats in a high-fat with cholesterol diet group had daily energy amounts provided by pellet feed mixed with 65.00% butter and 2.00% cholesterol. Before beginning the study and at the end, rats live weight was recorded and their blood samples were taken for biochemical analyses. Hematoxylin and Eosin and Crossman’s triple staining techniques were used to investigate the general structure of gastric tissue. Rats fed with HFCD had statistically significant increases in live weight and total cholesterol values, and were identified to have gastric tissue degeneration. The rats gastric tissue in control group had more intense somatostatin (SST) immunoreactivity in parietal and chief cells than the HFCD group. It was determined that feeding with the HFCD has a negative effect on SST secretion in rats and hence, this may have important areas of use such as in gastric cancer treatment and preventing complications linked to gastric diseases.

Abstract To the editor: We read the article entitled “Protective effects of Chromolaena odorata extract on experimental benign prostatic hyperplasia in rats” with great interest. This research aimed to evaluate the effects of hydro-methanol extract of Chromolaena odorata (HMECO) on testosterone propionate (TP)-induced benign prostate hyperplasia (BPH) rat model. We want to congratulate the authors for this original article and make some positive comments. The BPH is a common condition in both aged men and dogs. Although not considered a precursor of prostate cancer (PCa), BPH commonly affects the prostate gland, and shares some features with PCa, like symptoms, hormone-dependent growth and response to anti-androgen therapy.¹ Increasing our understanding in BPH can bring us more knowledge about PCa, for men and dogs. This is an article whose methodology is easy to replicate and whose authors know the specificities of this model of prostate hyperplasia. In our opinion, the remaining prostate lobes could have been evaluated, although in this specific model of BPH the hyperplasia of rat ventral prostate lobes is considered analogous to the morpho-logical alterations of human BPH. This article reinforces the importance of animal models in the preclinical evaluation of new therapies obtained from natural extracts. In a similar way, our research group evaluated effects of Castanea sativa Mill. flower (CF) in a N-methyl-N-nitrosourea (MNU) plus testosterone rat model.² Animals from induced groups received a multistep protocol for PCa induction, consisted of sequential administration of flutamide, testosterone propionate, the carcinogenic agent MNU and crystalline testosterone. Animals from treatment groups were exposed to CF extract in drinking water, at a dose of 3.00 mg per animal daily, for 49 weeks, starting at the time of the carcinogenesis induction. Animals were sacrificed at 61 weeks of age, approximately 10 months after MNU administration. Our results suggested that CF extract was well tolerated by the animals and did not cause severe hepatic or renal toxicity. Furthermore, the animals exposed to the CF extract showed fewer inflammation areas on the dorsolateral prostate lobe than those not exposed to the CF extract, suggesting that this extract may be used as chemopreventive agent against prostate cancer and seems to have an antioxidant role. In conclusion, the studies with animal models of BPH and PCa add value to the study of prostate diseases and to test the efficacy of natural compounds, and their extracts.  

Abstract In the present study the effect of ozone therapy on hydrofluoric acid (HFA) related eye burn was investigated in rats. A Total 20 healthy male Wistar albino rats (weighing 250 - 300 g with the age of 16 weeks) were used. They were divided into groups (experimental and control groups) of 10 rats being housed individually and fed ad libitum. The HFA (2.00%) burn was created in all animals. The ozonized (20.00 µg O₃ mL^-1) bi-distilled water was applied as a drop (10.00 µL each drop) every 8 hr for 7 days in the experimental group. At the same time, 0.90% NaCl was applied as drop (10.00 µL each drop) every 8 hr for 7 days in the control group. In the experimental group, intensive inflammation, angiogenesis, epithelial damage and stromal edema were detected in one animal. Epithelial vascularization and stromal edema were seen in four animals. In control group, only two animals’ corneal structures were normal. Inflammation, angiogenesis, epithelial damage, fibrosis, epithelial vascularization and stromal edema were detected in the rest. As a result of this study, it was observed that local usage of ozone therapy had a positive effect on the healing of corneal burns caused by HFA. It was concluded that more ozone-related studies should be done to enlighten the subject.     

Abstract Benign prostatic hyperplasia (BPH) is an age-related disease in dogs and man leading to prostate enlargement which impinges on the urethra causing urinary outflow obstruction. Due to the side effects of surgery and chemotherapy used for the treatment of this disease, attention is now focused on phytotherapeutics for its management. Thus, we investigated the inhibitory effect of hydro-methanol extract of Chromolaena odorata (HMECO) on testosterone propionate (TP)-induced BPH rat model. A total of forty-two 10-12 weeks old male Sprague-Dawley outbred albino rats (Rattus norvegicus) weighing 200 - 250 g were randomly divided into six equal groups of seven rats each based on body weight as follows: A) Control group given phosphate-buffered saline orally and corn oil subcutaneously (SC) once daily, B) TP at a dose of 3.00 mg kg^-1 SC once daily, C) TP at a dose of 3.00 mg kg^-1 SC and finasteride at a dose of 10.00 mg kg^-1 orally once daily, D) TP at a dose of 3.00 mg kg^-1 SC plus 200 mg kg^-1 HMECO orally once daily, E) TP at a dose of 3.00 mg kg^-1 SC plus 400 mg kg^-1 HMECO orally once daily and F) TP at a dose of 3.00 mg kg^-1 SC plus 800 mg kg^-1 HMECO orally once daily for 28 days. Results showed that HMECO significantly reduced prostate weight, prostatic index; serum levels of testosterone and prostatic epithelial thickness and increased luminal diameter in BPH induced rats. Thus, the results of this study suggest that C. odorata is a potential pharmacological candidate for the management of BPH.

Abstract Phthalates are environmental contaminants mostly used as plasticizers and additives in different products. Having endocrine-disrupting properties, phthalates are known as potential reproductive toxicants. The present study was conducted to evaluate the reproductive toxicity of di-n-butyl phthalate (DBP) in pregnant rats and their offspring and also to assess the ability of vitamin E in the elimination or reducing reproductive toxicity of DBP. Sixty-six pregnant Wistar rats were exposed to 100, 500 or 1,000 mg kg^-1 per day DBP or 500 mg kg^-1 per day DBP along with 100 mg kg^-1 per day vitamin E during gestation. After delivery, they were divided into two groups. In one group gavage was finished after litter while in the other DBP administration was continued till weaning. The results showed that DBP affected many aspects of reproductive performance in pregnant rats and their offspring. It could be suggested that vitamin E could ameliorate the adverse effects of DBP, especially in male pups.

Abstract Skin flap necrosis has been remained as an unsolved problem in plastic and reconstructive surgeries. Here, we explored the effects of metformin post-treatment on random skin flap survival in rats. An 8.00 × 2.00 cm dorsal skin flap was created in 24 rats and they were then divided into three groups (n = 8) including Control, metformin (Met) 50.00 mg kg^-1 and Met 100 mg kg^-1. All animals were administrated orally until seven days after flap surgery. Flap survival, the number of blood vessels and mast cells in the flap tissues were analyzed. Vascular endothelial growth factor (VEGF) expression levels in flap tissues was also determined using immunohistochemical methods. The percentage of survival area in Met 50.00 mg kg^-1 and Met 100 mg kg^-1 groups were significantly higher compared to control. The blood vessel density and the VEGF positive cells in the viable areas of flaps showed a significant increase in Met 50.00 mg kg^-1 group compared to control group. The results of this study suggested that treatment with metformin, especially with low dose following skin flap surgery was effective in improving the flap survival and increasing the neovascularization in the flaps tissues of rats.

Abstract Skeletal muscle atrophy induced by denervation is one of the common disorders in traumatic nerve injuries. The aim of this study was the evaluation of histomorphometrical changes of extensor digitorum longus muscle after denervation and its regeneration by tissue engineering. Ninety adult male Wistar rats were randomly divided into six main groups (n = 15) in three time periods (2, 4 and 8 weeks; n = 5). Control group was treated without surgery, in transection (Tr) group left sciatic nerve was transected, in scaffold (S) group only collagen gel scaffold was used, in mast cell (MC) group mast cells were used, mesenchymal stem cell (MSC) group was treated with mesenchymal stem cells and in MC+MSC group, mast cells along with mesenchymal stem cells were used. In the cellular groups, the scaffold and cells were mixed and placed in the transected nerve gap. The average diameter of muscle fibers, ratio of the muscle fibers nuclei to the fibrocytes nuclei (mn/fn), ratio of the muscle fibers nuclei number to the muscle fibers number (mn/mf), the average ratio of blood vessels to muscle fibers number (v/mf) and muscles weight in Tr group were the lowest compared to the other groups; but, in cellular and S groups, amelioration was observed according to the time period. However, in MC+MSC group, there were the highest ameliorative results. This study revealed that simultaneous use of MCs and MSCs mixed with collagen gel scaffold can be considered as a suitable approach to improve denervated skeletal muscle atrophy associated with sciatic nerve injury.

Abstract The study was done to ascertain the protective potentials of ethanol seed extract of Citrullus lanatus on aluminum chloride-induced reproductive and hematological toxicities. Thirty mature male rats were used for the study. They were assigned into five groups (n=6). Group1was treated daily with aluminum chloride (100 mg kg^-1) per os for 8 weeks. Group 2 was treated with aluminum chloride (100 mg kg^-1) and C. lanatus seed extract (CLSE) 200 mg kg^-1 per os simultaneously for 8 weeks. Group 3 was served as a normal control and given distilled water as a placebo per os daily for 8 weeks. Group 4 was only treated with CLSE (200mg kg^-1) for eight weeks. Group 5 was only treated with aluminum chloride (100 mg kg^-1) per os for 8 weeks and then treated with CLSE (200 mg kg^-1) per os for another 4 weeks. Testosterone level, testicular weight, sperm motility, gonadal sperm, and extragonadal sperm reserves showed significant increases in group 2 compared to groups 1 and 5. Optimum histoarchitectural protection of the seminiferous tubules was observed in group 2, which did not differ from normal ones. For the hematological parameters, optimum protection was also observed in group 2 compared to other groups. From the results, ethanol seed extract of C. lanatus demonstrated protective potentials against aluminum's harmful effects on the male reproductive system and hematology in an experimental male rat model.

Abstract Polyvinyl chloride (PVC) has toxic effects through the induction of oxidative stress in the body and testicles. Vitamin E (Vit E) is a dietary compound that functions as an antioxidant scavenging toxic free radicals. The present study aimed to probe the protective effect of Vit E against PVC-induced reprotoxicity in male rats. In this experimental study, 24 male rats were randomly divided into four groups (n=6) including control, Vit E (150 mg kg^-1 per day; orally), PVC (1000 mg kg^-1 per day; orally) and PVC + Vit E. After 40 days, rats were euthanized and epididymal sperms characteristics, embryo development and malondialdehyde (MDA) and testosterone levels were examined. The PVC decreased sperm count, motility and viability as well as testosterone level and increased sperms with damaged chromatin in comparison with controls. Also, the percentages of fertilization, two-cell embryos and blastocysts as well as MDA levels were decreased in PVC-treated rats. However, Vit E improved PVC-induced alterations in aforesaid parameters. The results indicated that PVC can reduce fertility potential in male rats probably through androgen and sperm quality and quantity reductions, while Vit E can exert protective effects in PVC-related reproductive toxicities.