The association between allergic rhinitis and airway dysfunction and nasal endothelial damage and oxidative stress.

BACKGROUND: Although lower airway hyperresponsiveness is present in approximately one in three patients with allergic rhinitis (AR), the underlying mechanism remains unclear. To evaluate nasal patency and pulmonary functions in AR independently of the presence of asthma and to investigate the relationships between these and nasal oxidative stress parameters and endothelial damage. METHODOLOGY: Seventy adolescents with AR (AR group - 27 with asthma and 43 without asthma) and 30 healthy controls (HC group) were included in this prospective, cross-sectional study. Endocan and oxidative biomarkers [total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI)] in nasal lavage fluid specimens; peak nasal inspiratory flow (PNIF); fractional exhaled nitric oxide (FeNO), and impulse oscillometry (zR5, zR20, and R5-20 for resistance and zX5 and zX20 for reactance) were investigated. RESULTS: Nasal endocan, TOS, and OSI values were higher in the AR group and TAS in the HC group. There was no difference between AR groups with and without asthma in terms of nasal endocan and oxidative biomarkers. FeNO levels and airway resistance (zR5, zR20, and R5-20) were higher in the AR group than in the HC group. However, there was no difference between the groups in PNIF. X5 was higher among the AR without asthma than in the other groups. Correlation between OSI and R5-20 was observed in the AR group. In the linear regression model, (logged) OSI was significantly predicted (logged) R5-20. CONCLUSIONS: The airways of adolescents with AR without asthma were as much affected as those of the AR with asthma, and this effect was associated with nasal endothelial damage and an increase in oxidative stress.

Long-term treadmill exercise upregulated hippocampal learning-related genes without improving cognitive behaviour in socially isolated rats.

BACKGROUND: Some environment enrichments such as exercise has been reported to improve the diminished cognitive functions and related gene expression. Therefore, we aimed to investigate the effects of prolonged treadmill exercise on long-term learning and hippocampal gene expression, which involves learning and plasticity. MATERIALS AND METHODS: Male Wistar rats (n = 32) randomly assigned into four groups: control (C), social isolation (SI), exercised (E), social isolation + exercise (SE) during postnatal days (PNDs) 21-34. Social isolation protocol was applied during 14 days by placing the rat alone in a cage. Rats were exercised daily, 5 days per week, for overall 4 weeks. Finally, learning performance was evaluated by the novel object recognition test. At the end of learning test, the rats were decapitated to isolate hippocampus tissues for learning related gene expression such as N-methyl-d-aspartate receptor (NMDAR) subunit genes (Grin1, Grin2a, Grin2b) and cyclin dependent kinase 5 (Cdk5), Cdk5 regulatory subunit p35 (Cdk5r), activity-regulated, cytoskeletal-associated protein (Arc), the immediate early gene (c-Fos, a marker of neuronal activation), doublecortin (DCX), achaetescute homolog 1 (ASCL1), brain-derived neurotrophic factor (BDNF) by real-time polymerase chain reaction (RT-PCR). RESULTS: Grin1, NMDAR subunit gene expression was increased significantly in E group compared to other groups. Grin2b, NMDAR subunit gene expression was increased in E group compared to the SI group. Cdk5 level increased in E group compared to the SE group. The ASCL1 gene expression increased in E group compare to the SE group. The DCX gene expression increasing in C group compared to SI and SE groups. CONCLUSIONS: Taken together these findings may point out that long-term social isolation down-regulated learning-related genes. However, treadmill exercise together with social isolation did not restore this down-regulation although treadmill exercise increased learning-related genes without improving cognitive behaviour.

Effects of platelet-rich plasma against experimental ischemia/reperfusion injury in rat testis.

BACKGROUND: Testicular torsion is a common problem and, to date, there is no agent to preserve testicular function following detorsion. Platelet-rich plasma (PRP), with its rich growth factor composition, has proven beneficial in regenerative therapy. It is believed that PRP has not been studied in testis for ischemia/reperfusion (I/R) injury. OBJECTIVE: This study investigated the effect of PRP in an I/R rat model 1 month after detorsion. STUDY DESIGN: Of 24 adult male Sprague-Dawley rats, 18 were randomly assigned into three groups, with six in each: control, I/R and I/R + PRP. The PRP was prepared from the remaining six. Each group underwent right orchiectomy. Ischemia was performed by rotating the left testis 720° and fixing with a nylon suture for 4 h. Reperfusion occurred 4 h later by removing the suture, and PRP was administered at a dose of 10 µl (2000 × 109/l) into the left testis via the intraparenchymal route. Animals were sacrificed at the fourth week, and testes were taken for malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), myeloperoxidase (MPO), transforming growth factor ß (TGF-ß), and caspase-3 measurements. RESULTS: Ischemia/reperfusion caused a significant increase in MDA, MPO and caspase-3 activity, and significant decrease in GSH levels and SOD activity. The PRP treatment helped correct the alterations in SOD, caspase-3, and MPO activities and MDA levels. However, the mean MDA level and MPO activity were not totally restored compared with the controls. Serum testosterone levels of the I/R group were significantly lower compared with the control and I/R + PRP groups. TGF-ß and caspase-3 protein expressions were significantly higher in the I/R group compared with the control group and were low with PRP administration compared with I/R groups (summary Table). DISCUSSION: The findings of the present study suggest that PRP, by inhibiting neutrophil infiltration and oxidative stress and increasing antioxidant defense, exerts protective effects on testicular tissues against I/R. This study had some limitations: a scoring system was not used in the assessment of spermatogenesis in the histopathological findings and specific testis cell types were not histologically assessed. CONCLUSIONS: In light of the biochemical, histological and, especially, hormonal findings, intraparenchymal PRP injection may have a protective effect in testicular tissue against I/R injury.

Quercetin protects radiation-induced DNA damage and apoptosis in kidney and bladder tissues of rats.

Ionizing radiation (IR) can induce cell damage and cell death through the reactive oxygen species generated by radiolytic hydrolysis. The present study was aimed to determine the possible protective effects of quercetin, a well-known antioxidant agent, against IR-induced bladder and kidney damage in rats. Sprague-Dawley rats were exposed to 8-Gy whole-abdominal IR and given either vehicle or quercetin (20 mg/kg, ip). Rats were decapitated at either 36 h or 10 days following IR, where quercetin or vehicle injections were repeated once daily, and kidney and bladder samples were obtained for the determination of myeloperoxidase and caspase-3 activities, an index of tissue neutrophil infiltration and apoptosis, respectively. Radiation-induced inflammation was evaluated through tissue cytokine, TNF-α levels. In order to examine oxidative DNA damage, tissue 8-hydroxydeoxyguanosine (8-OHdG) levels were measured. All tissues were also examined microscopically. In the saline-treated irradiation groups, myeloperoxidase and caspase-3 activities, 8-OHdG and TNF-α levels were found to be increased in both tissues (p < 0.05). In the quercetin-treated-IR groups, all these oxidant responses were prevented significantly (p < 0.05). The present data demonstrate that quercetin, through its free radical scavenging and antioxidant properties, attenuates irradiation-induced oxidative organ injury, suggesting that quercetin may have a potential benefit in radiotherapy by minimizing the adverse effects and will improve patient care.

Quercetin treatment against ischemia/reperfusion injury in rat corpus cavernosum tissue: a role on apoptosis and oxidative stress.

Reactive oxygen metabolites play an important role in the ischemia/reperfusion (I/R)-induced tissue injury. This study was designed to investigate the possible protective effects of quercetin against I/R injury of the rat corpus cavernosum tissue. To induce I/R injury, abdominal aorta was clamped for 30 min and reperfused for 60 min. Quercetin (20 mg/kg) or vehicle was given before ischemia and just after reperfusion in the I/R group and in the sham-operated control group in which clamping was not performed. After decapitation, corpus cavernosum tissues were removed and either placed in organ baths or stored for evaluating biochemical parameters. Oxidative injury was examined by measuring lucigenin chemiluminescence (CL), nitric oxide (NO), malondialdehyde (MDA) and glutathione (GSH) levels, superoxide dismutase (SOD) and myeloperoxidase (MPO) activities and caspase-3 protein levels. In the I/R group, contractile responses to phenylephrine and relaxation responses to carbachol were impaired significantly compared with those in the control groups, while quercetin treatment in I/R group reversed both of the responses. On the other hand, increase in lucigenin CL, NO, MDA levels and MPO and caspase-3 activities and decrease in GSH levels and SOD activity in the cavernosal tissues of the I/R group were also significantly reversed by quercetin treatment. Furthermore, observed distorted morphology with ruptured endothelial cells and vacuolization in the cytoplasm of cavernosal tissues of I/R no longer persisted in the quercetin-treated I/R group. Thus, our results suggested that treatment with quercetin may have some benefits in controlling I/R-induced tissue injury through its anti-inflammatory, anti-apoptotic, and antioxidant effects.

In vitro effects of nitric oxide donors on apoptosis and oxidative/nitrative protein modifications in ADP-activated platelets.

Nitric oxide (NO) is an important physiological signaling molecule. However, when produced in excessive amounts, NO can also have toxic effects. The aim of this study is to investigate the effects of exogenous- and endogenous-derived NO on oxidative modifications of proteins and apoptosis in activated platelets. Washed platelets were incubated with L-arginine or nitroso-glutathione (GSNO) in the presence of adenosine diphosphate (ADP). After incubation, caspase-3 activity, phosphatidylserine (PS) externalization and the potential of mitochondrial membrane as markers of apoptosis were measured. In addition, the alterations in protein carbonylation (PCO) and nitrotyrosine (NT) formation as markers of protein oxidation were examined. Platelet activation with ADP (20 µM) significantly increased PCO and NT levels and apoptotic events. After incubation with L-arginine, platelet NO production increased significantly. This L-arginine-induced increase caused decreases in formerly increased PCO and NT levels associated with ADP-induced platelet activation. Stimulation of NO production with L-arginine protected platelets from apoptosis. GSNO caused an increase in protein NT levels. Despite this change, GSNO was effective in inhibition of P-selectin expression, platelet aggregation, protein carbonylation and apoptosis. The results suggest that L-arginine and GSNO-mediated NO leads to the inhibition of key apoptotic processes including caspase-3 activation, PS exposure and low mitochondrial membrane potential in washed platelets. The inhibitory effect of platelet clearance of L-arginine and GSNO may be a novel useful therapeutic property in clinical application.

Influence of platelet γ-glutamyltransferase on oxidative stress and apoptosis in the presence of holo-transferrin.

Several studies have documented that formation of oxidant mediators may induce apoptosis in nucleated and anucleated cells by modulating intracellular signalling pathways. Reactive oxygen species (ROS) play a very important role in the platelet function. γ-Glutamyltransferase (GGT), a novel source of cellular production of oxidants in the presence of iron and reduced glutathione (GSH), is also found on platelets. The role of platelet-bound GGT in platelet apoptosis and oxidative stress is unknown. The aim of our study was to determine the effects of platelet GGT activity on oxidative stress and apoptotic events in vitro via determination of lipid peroxidation (LPO), protein oxidation, GSH, catalase, caspase-3 activation and phosphatidylserine (PS) exposure in the presence of holo-transferrin (Tf). Stimulation of platelet GGT activity with GSH and glycylglycine (GlyGly) increased caspase-3 activation and PS exposure. A significant increase in lipid and protein oxidation and decrease in GSH and catalase levels was also observed in platelets with stimulation of GGT activity in the presence of Tf. Inhibition of GGT activity effectively reduced all the markers. These results suggest that generation of ROS by the GGT/GSH/Tf system can modify the platelets' redox environment and induce apoptosis in in vitro conditions.

Oxidized-LDL and Fe3+/ascorbic acid-induced oxidative modifications and phosphatidylserine exposure in human platelets are reduced by melatonin.

Low-density lipoprotein (LDL) modifications and platelet activation are major risk factors for cardiovascular diseases. When platelets are exposed to oxidative stress, they become activated. Oxidized LDL (ox-LDL) and metal-catalysed oxidation systems such as Fe3+/ascorbic acid increase free radical production. We wanted to verify whether melatonin has a protective effect against oxidative modifications and phosphatidylserine externalization in platelets induced by ox-LDL and Fe3+/ascorbic acid. For in vitro effects of melatonin on platelets, ADP-activated platelets were incubated with ox-LDL or Fe3+/ascorbic acid for 1 h at 37 degrees C with or without melatonin. Then platelet malondialdehyde, protein carbonyl and glutathione levels were measured. Platelet phosphatidylserine exposure was measured with annexin-V using flow cytometry. Malondialdehyde, protein carbonyl and phosphatidylserine levels of platelets treated with Fe3+/ascorbic acid significantly increased compared to the control group. Glutathione contents of Fe3+/ascorbic acid-treated platelets significantly decreased. Melatonin pre-treatment of Fe3+/ascorbic acid-treated platelets caused a mar ked reduction in malondialdehyde and phosphatidylserine levels and a marked increase in glutathione levels. Melatonin also caused non-significant reduction in protein carbonyl contents of Fe3+/ascorbic acid-treated platelets. Malondialdehyde, protein carbonyl and phosphatidylserine levels of platelets treated with ox-LDL also significantly increased compared to the control group. Platelet glutathione levels non-significantly decreased with ox-LDL. With addition of melatonin, malondialdehyde, protein carbonyl and phosphatidylserine levels of platelets treated with ox-LDL significantly decreased. These data suggest that melatonin may protect platelets from iron overload-induced and ox-LDL-induced oxidative modifications and also from the triggering signals of apoptosis activation, possibly due to its scavenger effect on toxic free radicals.