Short term effects of experimental gastric outlet obstruction and truncal vagotomy on gut hormones.

Gastric outlet obstruction (GOO) is caused mainly by pyloric or duodenal blockage; gastric surgery and vagotomy are effective treatments. We investigated the short term effects of experimental GOO and truncal vagotomy (TV) on gut hormone levels. We used 8-week-old male Wistar rats divided randomly into four groups: control, GOO, TV, and GOO + TV. At the end of the experiment, blood and tissue samples of the pylorus and fundus were obtained for biochemical and immunohistochemical analysis. Gastric motility decreased in the TV group, but there was no difference in food intake compared to the control group; water consumption and urine output were increased. Feces excretion and food intake decreased due to loss of food movement from the stomach of GOO and GOO + TV rats. Levels of insulin and ghrelin were lower than for the control group, but levels of cholecystokinin were higher. Leptin and glucagon-like peptide 1 levels were increased in the GOO group, while somatostatin was decreased. Leptin immunostaining levels were decreased in the GOO + TV group. Gastrin and neuropeptide Y levels were lower in the GOO and GOO + TV groups compared to the other groups. We found that both gut hormone levels related to gastric motility and metabolism, and immunohistochemical staining of the stomach tissue were altered by TV and GOO. Measuring changes in gut hormones following gastric surgery could be useful for monitoring the effectiveness of treatment.

Investigation of Neuroprotective and Therapeutic Effects of Hesperidin in Experimental Spinal Cord Injury.

AIM: To investigate the neuroprotective and therapeutic efficacy of hesperidin against secondary damage following traumatic spinal cord injury. MATERIAL AND METHODS: A total of 32 male Wistar albino rats weighing 250?300 g were randomly divided into four groups (n=4): group I, control group; group II, sham group; group III, preconditioning group, and group IV, treatment group. A rat model of spinal cord injury was established by dropping a weight of 100 g/cm on the spinal cord exposed at T7?T10 with dorsal laminectomy. In neurological examination after the trial period, inclined planed test, modified Tarlov scale, and finger extension test were performed. Furthermore, the bioefficacy of hesperidin was investigated histopathologically, biochemically, and immunohistochemically using blood and tissue samples obtained from the experimental animals. RESULTS: Neurological examination following spinal cord injury revealed that hesperidin significantly contributed to improvement in the 24-hour period. Biochemical analyses revealed that hesperidin showed anti-inflammatory effects by decreasing IL-1? and TNF-? levels at the 24th hour as well as strong antioxidant activity by increasing TAS levels in groups III and IV. Histopathologically, hesperidin reduced hemorrhage, laceration, axonal and neuronal degeneration, necrosis, inflammatory reaction, and edema in groups III and IV. Immunohistochemically, hesperidin reduced the number of caspase 3-positive apoptotic cells in groups III and IV. CONCLUSION: Hesperidin showed antioxidant, anti-inflammatory, and anti-apoptotic effects during the acute period following spinal cord injury; thus, hesperidin shows neuroprotective and therapeutic efficacy in spinal cord injury.

Antioxidant Effects of Oleuropein on Hydrogen Peroxide-Induced Neuronal Stress- An In Vitro Study.

BACKGROUND: Persistent oxidative stress can lead to chronic inflammation and mediate most chronic diseases including neurological disorders. Oleuropein has been shown to be a potent antioxidant molecule in olive oil leaf having antioxidative properties. OBJECTIVE: The aim of this study was to investigate the protective effects of oleuropein against oxidative stress in human glioblastoma cells. METHODS: Human glioblastoma cells (U87) were pretreated with oleuropein (OP) essential oil 10 µM. After 30 minutes, 100 µM H2O2 was added to the cells for three hours. Cell survival was quantified by colorimetric MTT assay. Glutathione level, total oxidant capacity, total antioxidant capacity and nitric oxide levels were determined by using specific spectrophotometric methods. The relative gene expression level of iNOS was performed by qRT-PCR method. RESULTS: According to viability results, the effective concentration of H2O2 (100µM) significantly decreased cell viability and oleuropein pretreatment significantly prevented the cell losses. Oleuropein regenerated total antioxidant capacity and glutathione levels decreased by H2O2 exposure. In addition, nitric oxide and total oxidant capacity levels were also decreased after administration of oleuropein in treated cells. CONCLUSION: Oleuropein was found to have potent antioxidative properties in human glioblastoma cells. However, further studies and validations are needed in order to understand the exact neuroprotective mechanism of oleuropein.

Low concentration of oleic acid exacerbates LPS-induced cell death and inflammation in human alveolar epithelial cells.

PURPOSE: The current study aimed to investigate in vitro effects of oleic acid on lipopolysaccharide (LPS)-induced acute lung injury in the human lung epithelial cells (A549). MATERIALS AND METHODS: The cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tests. Selected gene expression levels were analyzed by Real-Time Quantitative-Polymerase Chain Reaction (RT-qPCR). RESULTS: 24 hours of LPS (100 ng/mL) exposure decreased the cells' viability by 44.6% compared to untreated control. Low concentration (2.5 nM) of oleic acid slightly suppressed the cell survival by 9.1% analyzed 24 hours after incubation. However, oleic acid pretreatment before LPS exposure significantly increased cell survival loss to 63.9%. LPS exposure decreased the expressions of catalase (CAT) and glutathione peroxidase (GPx) mRNA levels by 2.8 and 2.5 fold, respectively. Moreover, pretreatment of the cells with oleic acid strengthened LPS-decreased expressions of CAT and GPx genes by 3.5 and 6.7 fold, respectively. The mRNA expressions of superoxide dismutase (SOD), induced nitric oxide synthase (iNOS), interleukin-1ß, IL-12, COX-2, caspase-3 and caspase-8 were increased by 2.4, 2.2, 2.2, 2.3, 3.0, 2.6, and 2.5 fold, respectively, by LPS, and oleic acid pretreatment significantly potentiated the effect of LPS. CONCLUSION: Oleic acid worsens LPS-induced cell death by potentiating oxidative stress and inflammation in A549 lung epithelial cells.

Beneficial effects of nontoxic ozone on H2O2-induced stress and inflammation.

In this study, the anti-oxidant and anti-inflammatory efficacy of ozone oxidative preconditioning (OOP) were investigated on hydrogen peroxide (H2O2)-induced human lung alveolar cells. In MTT and trypan blue viability tests, while 100 µmol/L H2O2 caused a 17.3% and 21.9% decrease in the number of living cells, respectively, ozone at 20 µmol/L regenerated cell proliferation and prevented 9.6% and 11.0% of cell loss, respectively. In addition, H2O2 decreased the transcription levels of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD) 5.43-, 2.89-, and 5.33-fold, respectively, while it increased Bax, NF-κß, TNF-α, and iNOS expression 1.57-, 1.32-, 1.40-, and 1.41-fold, respectively. Ozone pretreatment, however, increased CAT, GPx, and SOD transcription levels 7.08-, 5.17-, and 6.49-fold and decreased Bax, NF-κß, TNF-α, and iNOS transcriptions by 1.25-, 0.76-, 3.63-, and 7.91-fold, respectively. Moreover, intracellular glutathione (GSH) level and SOD activity were decreased by 46.2% and 45.0% in the H2O2 treatment group, and OOP recovered 58.5% and 20.1% of the decreases caused by H2O2. H2O2 also increased nitrite levels 7.84-fold, and OOP reduced this increase by half. Consequently, OOP demonstrated potent anti-oxidant and anti-inflammatory effects on in vitro model of oxidative stress-induced lung injury.

Silencing of PrP C (prion protein) expression does not affect Brucella melitensis infection in human derived microglia cells.

Cellular prion proteins (PrP(C)) are mainly expressed in the central nervous system where they have antioxidant effects and a role in the endocytosis of bacteria within cells. These proteins also have some crucial biological functions including roles in neurotransmission, signal transduction and programmed cell death. However, the role of prion proteins in neuronal Brucella infection, specifically in the interaction of the pathogen and the host cell is controversial. In the present study, the silencing of PrP(C) mRNA by small interfering RNA (siRNA) transfection was investigated in human microglia cells infected with Brucella melitensis. More than 70% of prion proteins were down-regulated in microglia by siRNA transfection and this caused a slight decrease in the cellular viability of the control cells. Silencing of PrP(C) suppressed the antioxidant systems, though it led to an up-regulation of pro-inflammatory cytokines such as IL-12 and TNF-α as demonstrated by qRT-PCR analysis. B. melitensis infection of prion protein-silenced cells led to increase host viability, but had no effect on bacterial phagocytosis. According to the present study, there is no significant effect of prion proteins on phagocytosis and intracellular killing of B. melitensis in microglia cells.

Cisplatin reduces Brucella melitensis-infected cell number by inducing apoptosis, oxidant and pro-inflammatory cytokine production.

Brucella species are able to survive and replicate within the phagocytic cells and cause chronic infections in domestic animals and humans. Modulation of programmed cell death by Brucella spp. may be one of the reasons of the chronicity of the infection. In this study, whether cisplatin treatment, an apoptotic anticancer agent, would enhance the host resistance against Brucella melitensis-infected human macrophage-like cells was investigated. The infection neither induced inflammation nor oxidative stress. But, Brucella caused a decrease in infected macrophage viability of 36% at 48 h postinfection (p.i.) as compared with uninfected cells. Treatment of infected macrophages with 20 microM cisplatin for 48 h caused a large increase in nitric oxide (NO) levels in a time-dependent manner via induction of iNOS transcription. Cisplatin also enhanced glutathione peroxidase, myeloperoxidase and xanthine oxidase activities, providing evidence of generation of reactive free radicals. N-acetylcysteine was able to decrease cisplatin-induced NO, and prevented the agent-induced apoptosis, similar to effects found in l-NAME (N(G)-nitro-l-arginine methyl ester) treatment. Cisplatin stimulated inflammation through the induction of TNF-alpha and IL-12 secretion, and down-regulated Brucella-stimulated IL-10 transcription. The number of infected cells and their viability were decreased by 80% at 48 h p.i. by cisplatin in comparison with infected cells. Similar to this result, cisplatin treatment resulted in reduced intracellular CFU of B. melitensis being reduced by 80% at 48 h p.i. These findings demonstrate that pharmacological agents such as cisplatin may be considered to influence immune responses and apoptosis to help decrease Brucella-infected cell number.