Evaluation of the effects of empagliflozin on acute lung injury in rat intestinal ischemia-reperfusion model.

BACKGROUND: Empagliflozin is a selective sodium-glucose co-transporter (SGLT2) inhibitor that is approved for the treatment of type 2 diabetes. The beneficial effects of empagliflozin on other organ systems including the heart and kidneys have been proven. The aim of this study is to evaluate the role of empagliflozin on acute lung injury induced by intestinal ischemia-reperfusion (I/R). MATERIALS AND METHODS: A total of 27 male Wistar albino rats were divided into three groups: sham, I/R, and I/R + empagliflozin; each group containing nine animals. Sham group rats underwent laparotomy without I/R injury. Rats in the I/R group underwent laparotomy, 1 h of after ischemia-reperfusion injury (superior mesenteric artery ligation was followed by 2 h of reperfusion). Rats in I/R were given empagliflozin (30 mg/kg) by gastric gavage for 7 days before the ischemia-reperfusion injury. All animals were killed at the end of reperfusion and lung tissue samples were obtained for immunohistochemical staining and histopathological investigation in all groups. RESULTS: Serum glucose, AST, ALT, creatinine, native thiol, total thiol, and disulfide levels and disulfide-native thiol, disulfide-total thiol, and native thiol-total thiol ratios as well as the IMA levels were analyzed and compared among the groups. While intestinal I/R significantly increases serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and creatinine levels; did not cause any change in homeostasis parameters and IMA level. Empagliflozin treatment had no significant effect on biochemical parameters. Empagliflozin treatment induced a significant decrease in positive immunostaining for IL-1, IL-6, TNF-alpha, caspase 3, caspase 8, and caspase 9 compared to the I/R group in lung tissue samples. Intestinal I/R caused severe histopathological injury including edema, hemorrhage, increased thickness of the alveolar wall, and infiltration of inflammatory cells into alveolar spaces. Empagliflozin treatment significantly attenuated the severity of intestinal I/R injury. CONCLUSIONS: It was concluded that empagliflozin treatment may have beneficial effects in acute lung injury, and, therefore, has the potential for clinical use.

The Cytokines-Directed Roles of Spirulina for Radioprotection of Lacrimal Gland.

PURPOSE: To evaluate the radioprotective effect of spirulina (SP) on the lacrimal glands after RAI treatment. METHODS: A total of 30 rats were separated into control, RAI and SP group. The radioprotective effect of SP on lacrimal glands was evaluated with histopathological and cytopathological analysis. Lacrimal glands were analyzed for tumor necrosis factor alpha (TNF-α), interleukin-2 (IL-2), IL-4, IL-6, IL-10, nuclear factor-kappa B (NF-κB), total oxidant status (TOS) and total antioxidant capacity (TAC) levels. RESULTS: RAI increased TNF-α (p = .001), IL-6 (p = .018), and NF-κB levels (p < .0005). Following the administration of SP, TNF-α (p < .0005), IL-4 (p = .026), and IL-6 (p = .006) levels decreased. RAI decreased the TAC levels (p = .001), and co-administration of SP increased the TAC level, but was not statistically significant. SP decreased the TOS level after RAI (p = .022)                . CONCLUSIONS: SP protects lacrimal glands from RAI-induced damage.

Protective role of silibinin over nickel sulfate-induced reproductive toxicity in male rats.

Nickel damages the male reproductive system. We investigated the beneficial effects of silibinin which has metal-chelating and antioxidant properties over nickel toxicity. Both antioxidative effects in testes and overall effects related to sperm motility, membrane and acrosome integrity of orally administered Silibinin were evaluated against the harmful effects of 30 day of intraperitoneal nickel sulfate (5 mg/kg/day) administration in rats. Male rats were randomized into control (Group1; n=6) and three experimental groups (n=6, each): Group2 Nickel sulfate (5 mg/kg/day), Group3 Silibinin (150 mg/kg/day), and Group 4 Nickel sulfate (5 mg/kg/day) + Silibinin (150 mg/kg/day). We found higher sperm motility, viable sperm and total sperm count in Groups 3 and 4 than the Group 2 treatment groups and the percentage of abnormal spermatozoa was similar in both groups (Groups 2 and 4). Increased apoptosis, activation of caspase3, 8, 9 and TUNEL were detected in Group 2. However, activation of caspase3, 8, 9 and TUNEL was reduced in Group 4. The protective effects of silibinin were demonstrated on histopathologic findings and some sperm parameters (sperm motility percentage, viable spermatozoa, sperm count, and abnormal spermatozoa percentage) in rats exposed to nickel.

Beneficial effects of vitamin E on radioiodine induced gastrointestinal damage: an experimental and pathomorphological study.

OBJECTIVES: The aim of the present study was to investigate the radioprotective effect of vitamin E in the prevention of radioiodine (RAI) induced gastrointestinal damage. METHOD: Twenty-four rats were randomly divided into three groups as follows: Group-1 was untreated control group, Group-2 was orally administered single dose of 111 MBq RAI, and Group-3 was orally administered 111 MBq RAI and 1 mL of oral vitamin E. Vitamin E was started two days before RAI administration and was continued for five days once daily after RAI. Pathomorphological parameters of gastrointestinal tissues (stomach, small intestines and bowels) were measured using Hematoxylin-Eosin and Masson's trichrome staining. RESULTS: Varying degrees of inflammation, edema, ulcer, mucosal degeneration, necrosis and fibrosis were seen in the stomach, small intestine and bowel tissues of the rats in both study groups and not in the control group. The differences were statistically significant between these groups for all parameters (p < 0.05). The histopathological damage in the vitamin E treated group was significantly less than the damage in the RAI only group (p < 0.05 for all pathomorphological parameters). CONCLUSION: The results of this study showed that vitamin E has a radioprotective property with antiinflammatory and antifibrotic effects protecting against gastrointestinal damage caused by radioiodine (Tab. 3, Fig. 3, Ref. 26).

Expression of ADAMTS-7 in myocardial dystrophy associated with white muscle disease in lambs.

The aim of the present study was to investigate the role of ADAMTS-7 gene in the pathogenesis of myocardial dystrophy associated with white muscle disease (WMD) in lambs. A total of 217 cardiac tissue samples from lambs with WMD were used in the study. Histopathological sections of the samples were stained with hematoxylin-eosin (HE) and examined using Western-blot, real-time PCR (RT-PCR) and immunohistochemistry for ADAMTS-7 gene expression, and the findings were statistically evaluated. Histopathological examinations revealed fibrosis associated with hyalinization, necrosis and granular calcifications in cardiomyocytes. Western blot and RT-PCR showed a statistically significant upregulation of ADAMTS-7 (p<0.05) (p<0.05). Immunohistochemical analyses showed that immunopositive cell numbers significantly high for ADAMTS-7 (p<0.05). The study has revealed that ADAMTS-7 gene is significantly expressed in myocardial dystrophy associated with WMD in addition to its role in the pathogenesis of this disease.

The protective effect of melatonin on sperm quality in rat after radioiodine treatment.

This study was designed to investigate the potential radioprotective impact of melatonin on the testicular tissue and sperm quality in rat given radioactive iodine (RAI) therapy. Thirty-six male Wistar albino rats were randomly divided into three groups as untreated control (Group 1); oral radioiodine group (RAI, 111 MBq, administrated rats); and RAI+melatonin group (oral radioiodine and intraperitoneal 12 mg/kg/day melatonin, starting 2 days before and continuing for 1 week after oral RAI administration). Twenty-four hours after the injection of the last melatonin dose, blood samples were taken for hormone analyses and the determination of the total antioxidant capacity. Sperm samples taken from the cauda epididymis were examined for spermatological parameters. Tissue samples taken from the rat testes were stained by TUNEL assay and with haematoxylin-eosin to detect apoptosis and histological alterations. It was demonstrated a significant decrease in epididymal spermatozoa viability and motility in all of the treatment groups, in comparison with the control group (p < .001). A significant decrease was also detected in sperm DNA fragmentation, follicle-stimulating hormone (FSH) level and the index of apoptotic germ cells in the RAI+melatonin group when compared to the radioiodine group. It was concluded that melatonin prevents the adverse affects of RAI on apoptosis and spermatozoa quality.

Evaluation of intra-testicular injections of calcium chloride and 4-vinylcyclohexene 1,2 monoepoxide for chemical sterilization in guinea pigs.

This study was aimed at investigating the use of intra-testicular calcium chloride (CaCl2) and 4-vinylcyclohexene 1,2-monoepoxide (VCM) injections as a side effect-free alternative method for the control of reproduction in guinea pigs. Fifty male guinea pigs were randomly assigned to five groups. In all groups, the chemical agents were injected into both testes in 1% lidocaine hydrochloride. While Groups I, II and III were administered with a single dose (0.25 mL) of sterile physiological saline, 15 mg/100 g CaCl2, and 240 mg/kg VCM, respectively, Group IV and V received a daily dose of 15 mg/100 g CaCl2, and 240 mg/kg VCM for 3 days, respectively. On day 90 post-administration, all animals were weighed and later decapitated under ether anaesthesia. Blood and tissue (testis, liver, hypophysis and adrenal gland) samples were taken. Sperm samples from the cauda epididymis were examined for spermatological parameters. Blood was used for hormone analyses and tissue samples were examined histopathologically (haematoxylin-eosin) and immunohistochemically (Tunel staining). The epididymal sperm count decreased in all treatment groups. Excluding 2 animals, Group V displayed azoospermia. When compared to the control group, Group V displayed the highest prolactin and lowest testosterone levels, and Group III showed the highest testosterone level. Histopathological examination revealed no intoxication finding. Chemical castration with VCM may be a good alternative to surgical castration as it enables mass sterilization without postoperative risks in guinea pig.

Melatonin: a hepatoprotective agent against radioiodine toxicity in rats.

OBJECTIVES: The aim of the current study was to investigate the possible radioprotective effects of melatonin against hepatic radioiodine (RAI) toxicity. METHODS: Thirty-six rats were randomly divided into three groups: untreated control (Group 1); oral radioiodine (RAI, 111 MBq) administrated rats (Group 2), and melatonin group (oral RAI and daily intraperitoneal injection of 12 mg/kg melatonin-Group 3). In the third group, melatonin administration was started two days before and continued for five days after RAI administration. Twenty-four hours after the administration of the last dose of melatonin, liver samples were taken for biochemical and histopathological evaluation. RESULTS: Oxidative stress parameters demonstrated that melatonin treatment decreased the tissue malondialdehyde (MDA), advanced the oxidation protein products (AOPP) levels, and increased the total-SH (sulphydryl) levels when compared with RAI group. The differences were statistically significant between these groups for all parameters (p < 0.05). The histopathological damage in the melatonin-treated group was significantly less than the damage in RAI group (p < 0.05 for all pathological parameters). CONCLUSION: The results of this study demonstrated that melatonin reduced the harmful effects of RAI treatment on the liver. Anti-inflammatory and antioxidant activities are likely to be involved in the mechanism underlying the radio-protective effects of melatonin (Tab. 3, Fig. 1, Ref. 30).

Effects of epidermal growth factor on reduction of the formation of thrombus and vessel wall healing in an experimental rat model.

OBJECTIVES: The aim of the current study was to investigate the synergistic effects of epidermal growth factor (EGF) and Enoxaparin in thrombus resolution. METHOD: Forty rats were divided into five groups (n = 8/group). Thrombosis was induced in all groups except the Sham group. Group 1: Sham; Group 2: Phosphate buffered saline; Group 3: Enoxaparin; Group 4: EGF; Group 5: EGF+Enoxaparin. The treatments were applied 2 hours preoperatively, then postoperatively at 48 hours. Rats were sacrificed 7 days after the 2nd injection. Tissue samples were examined with hematoxylin-eosin, trichrome, vascular endothelial growth factors (VEGF), von Villebrand factor (VWF), CD34 and CD68 for histopathological and immunohistochemical analyses. RESULTS: Neovascularisation, recanalization and macrophage accumulation were statistically significantly higher in the EGF+Enoxaparin group than the other groups (p < 0.05), and the volume of thrombus was determined to be significantly lower. Recanalization was found to be higher in the Enoxaparin group than in the other groups. As for the thrombus resolution, statistically significant regress in the EGF+Enoxaparin group (p < 0.05) compared to the other groups was found. Immunohistochemical antibodies were statistically higher in the EGF+Enoxaparin group than in the other groups (p < 0.05). CONCLUSION: The results of this study demonstrate that concomitant use of EGF and Enoxparin has a synergistic effect and contributes significantly to thrombus resolution (Fig. 10, Ref. 35).

Ultrastructural investigation of the protective effects of propolis on bleomycin induced pulmonary fibrosis.

We investigated the antioxidant and anti-inflammatory effects of propolis on bleomycin induced lung fibrosis and compared these effects to prednisolone treatment. Forty rats were divided into four groups of ten: group 1 was treated with intratracheal infusion of 0.2 ml physiological saline followed by daily treatment with 0.5 ml physiological saline for 20 days. In the remaining groups (groups 2 - 4), 5 mg/kg bleomycin was given via the trachea. Rats in group 2 were given 0.5 ml physiological saline. Rats in group 3 were treated with 100 mg/kg propolis, and 10 mg/kg prednisolone was given to rats in group 4. The treatments for all groups were continued for 20 days. On postoperative day 21, blood and lung samples were taken for biochemistry, histopathology and electron microscopy evaluation. We compared oxidative stress parameters and found lower malondialdehyde and myeloperoxidase levels, and higher total sulfhydryl levels and catalase activities for the bleomycin + propolis group than for the bleomycin and bleomycin + prednisolone groups. The highest mean fibrosis score was detected in the bleomycin group. Although the mean fibrosis scores of the bleomycin + propolis and bleomycin + prednisolone groups were not significantly different, electron microscopy revealed that propolis diminished bleomycin induced lung fibrosis more effectively than prednisolone. The effects of propolis might be due to its potent antioxidant and anti-inflammatory properties.

The protective effect of L-carnitine against hippocampal damage due to experimental formaldehyde intoxication in rats.

We investigated the protective effects of L-carnitine on hippocampus tissue damage in rats during experimental formaldehyde (FA) intoxication. Male Wistar albino rats were assigned into four groups: (1) control (C), (2) formaldehyde (FA), (3) formaldehyde + 0.5 g/kg of L-carnitine (FA + 0.5 LC) (4) formaldehyde + 1 g/kg L-carnitine (FA + 1 LC). At the end of the 14 day trial period, animals were sacrificed by decapitation under anesthesia. The hippocampus tissue samples were extracted to measure MDA, GSH and SOD activity. Neuronal degeneration was assessed based on histopathological (hematoxylin and eosin) and immunohistochemical (anti-ubiquitin) examination. To detect oxidative stress, specimens were reacted with anti-Cu/Zn-SOD antibody. After administering L-carnitine with FA to the animals, the activities of SOD and GSH increased, but the levels of MDA decreased in hippocampus tissue. Neuronal degeneration was observed in the FA group. L-carnitine administration reduced neuronal degeneration and histological structure was similar to controls. After FA application, degenerated hippocampus neurons were stained with anti-ubiquitin and Cu/Zn-SOD antibodies; weakly positive staining was observed in L- carnitine-treated groups. L-carnitine may be useful for preventing oxidative damage in the hippocampus tissue due to formaldehyde intoxication.

Radioprotective effect of montelukast sodium in rat lacrimal glands after radioiodine treatment.

PURPOSE: The aim of this study was to evaluate the morphological changes of rat lacrimal glands at the third month following radioiodine (RAI) treatment and the radioprotective effect of montelukast (ML) sodium against RAI-related lacrimal gland damage. METHODS: Fifty female Wistar Albino rats were divided into three groups. The control group (n=10) consisted of rats with no intervention. RAI group (n=20) consisted of rats treated with oral (131)I (111 MBq). The ML group (n=20) consisted of rats treated with intraperitoneal 10mg/kg/day ML sodium, starting three days before and continuing for one week after oral RAI administration. Intraorbital (IG), extraorbital (EG) and Harderian glands (HG) were removed bilaterally after three months. RESULTS: The existence of acinar atrophy, acinar fibrosis, abnormal cell lines, peripheral basophilia, cell size variation and decrease in amount of cytoplasm was significantly more common in the RAI-rat treated group than in the ML group, in each of the glands. The ML-treated group had less-frequent cell shape variation in EG (P=0.001) and HG (P=0.027), cell size variation in IG (P<0.001) and HG (P=0.01), ductal pathology in EG (P<0.001) and HG (P<0.001) and lipofuscin accumulation in EG (P=0.001) and in HG (P=0.01) than the RAI-treated group. CONCLUSIONS: RAI treatment seems to cause morphological damage to rat lacrimal glands, and ML sodium effectively protects against damage to lacrimal glands.