Treatment with Rutin - A Therapeutic Strategy for Neutrophil-Mediated Inflammatory and Autoimmune Diseases: - Anti-inflammatory Effects of Rutin on Neutrophils.

OBJECTIVES: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor (TNF)-α productions and MPO activity in human neutrophils were investigated. METHODS: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin (25 µM) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the TNF-α, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin (1 - 100 µM), after which MTT was appended and incubated at 37ºC for 4 hour. RESULTS: Rutin at concentrations up to 100 µM did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and TNF-α productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). CONCLUSION: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and TNF-α productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/ autoimmune diseases.

Anti-inflammatory Effects of Quercetin and Vitexin on Activated Human Peripheral Blood Neutrophils: - The effects of quercetin and vitexin on human neutrophils.

OBJECTIVES: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor α (TNF-α), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), TNF-α, and MPO productions in human neutrophils. METHODS: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin (1 - 100 µM) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin (25 µM) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) (10-7 M). NO production was carried out through nitrite determination by using the Griess method. Also, the TNF-α and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. RESULTS: Neutrophil viability was not affected up to a concentration of 100 µM of quercetin or vitexin. Both quercetin and vitexin significantly inhibited TNF-α, NO, and MPO productions in human neutrophils (P < 0.001). CONCLUSION: The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

The Effects of Kainic Acid-Induced Seizure on Gene Expression of Brain Neurotransmitter Receptors in Mice Using RT2 PCR Array.

INTRODUCTION: Kainic acid (KA) induces neuropathological changes in specific regions of the mouse hippocampus comparable to changes seen in patients with chronic temporal lobe epilepsy (TLE). According to different studies, the expression of a number of genes are altered in the adult rat hippocampus after status epilepticus (SE) induced by KA. This study aimed to quantitatively evaluate changes in the gene expression of brain neurotransmitter receptors one week after administration of kainic acid in the mouse hippocampus. METHODS: We used 12 BALB/c mice in this study and randomly divided them into 2 groups. To both groups, saline (IP) was administered for 7 days, and on the last day, KA (10 mg/kg, IP) was injected 30 minutes after administration of saline. Subsequently, behavioural changes were observed in mice. Then, in one group (1 day group), 2 hours and in another group (7 days group), 7 days after KA administration, the hippocampus tissue of mice was removed and used for gene expression analyses. Total brain RNA was isolated and reversely transcribed. We performed qPCR using RT2 Profiler TMPCR Array Mouse Neurotransmitter Receptors and Regulators (QIAGEN) containing primers for 84 genes. In this regard, we selected 50 related genes for KA model. RESULTS: Our results showed significant changes in the gene expression of GABAA subunits receptors, including α1-α3, α5, α6, ß2, ß3, γ1, ρ, and rho1-2 on day 7 compared with the day 1. CONCLUSION: Expression of both inhibitory and excitatory receptors changed after one week. Further studies are needed to find more molecular changes in the gene expression of brain neurotransmitter receptors and regulators over longer periods of time in KA models using RT2 PCR array.

Comparison between the effects of quercetin on seizure threshold in acute and chronic seizure models.

Flavonoids are important constituents of food and beverages, and several studies have shown that they have neuroactive properties. Many of these compounds are ligands for γ-aminobutyric acid type A receptors in the central nervous system. This study aimed to investigate the anticonvulsant effects of quercetin (3,3',4',5,7-pentahydroxyflavone), which is a flavonoid found in plants, in rats treated with pentylenetetrazole in acute and chronic seizure models. Single intraperitoneal administration of quercetin did not show anticonvulsive effects against acute seizure. Similarly, multiple oral pretreatment with quercetin did not have protective effects against acute seizure. However, multiple intraperitoneal administration of quercetin (25 and 50 mg/kg) significantly increased time to death compared with the control (p < 0.001). However, quercetin pretreatment had no significant effects on the pattern of convulsion development during all periods of kindling. But on the test day, quercetin (100 mg/kg) could significantly increase generalized tonic-clonic seizure onset (GTCS) and decrease GTCS duration compared with the control (p < 0.01, p < 0.05). We conclude that quercetin has a narrow therapeutic dose range for anticonvulsant activities in vivo, and it has different effects on the seizure threshold. The different effects of quercetin on seizure threshold may occur through several mechanisms.

Report of Severe Menorrhagia Following the Maximum Amount of Lamotrigine Overdose.

Lamotrigine is an antiepileptic drug used as a treatment for partial and generalised seizures as well as for bipolar disorder type I. Till date, very few cases of lamotrigine overdose have been reported. The spectra of clinical effects of lamotrigine in acute overdose are not well established. In severe cases of poisoning, serious effects such as coma, respiratory depression, recurrent seizures and intraventricular conduction disturbances have been noted. Here, we report a case of lamotrigine overdose in a 26-year-old divorcee with paradoxical seizure activity and coma. On admission, the patient had a reduced level of consciousness. Serum evaluation revealed high lamotrigine levels without any other aetiology for mental dysfunction. To the best of our knowledge, this is the first report to describe a patient overdosed with 40 g of lamotrigine alone, which is the highest amount of lamotrigine overdose reported so far. During hospitalisation, the patient's haemoglobin level reduced from 12.9 to 7.7 g/dl and potassium level decreased repeatedly. More importantly, severe menorrhagia was noted. Following prompt supportive treatment with early intubation, along with the use of potassium chloride for hypokalaemia and administration of sodium bicarbonate, the patient's conditions improved and she was discharged from the hospital after 13 days.

Rutin activates the MAPK pathway and BDNF gene expression on beta-amyloid induced neurotoxicity in rats.

Flavonoids are present in foods such as fruits and vegetables. A relationship between the consumption of flavonoid-rich foods and prevention of human disease including neurodegenerative disorders has been demonstrated. We assessed the effect of rutin (3,3',4',5,7-pentahydroxyflavone-3-rhamnoglucoside) on the mitogen-activated protein kinase (MAPK) pathway, memory retrieval and oxidative stress in rats injected with ß-amyloid (Aß), which is implicated to have an important role in Alzheimer's disease (AD). Aß was injected bilaterally in the deep frontal cortex of rat brain. Next, rutin and saline were injected (i.p.) for 3 weeks. In comparison to the control group, rutin significantly increased extracellular signal-regulated protein kinase 1 (ERK1), cAMP response element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) gene expression in the hippocampus of rats. Rutin (100 mg/kg) significantly increased memory retrieval compared to the control group. Malondialdehyde (MDA) level in the hippocampus of the rutin group was significantly lower than those in the control group. The content of sulfhydryl groups in the rutin group was higher than that in the control group. The findings show a possibility that rutin may have beneficial effects against neurotoxicity of Aß on memory in rats.

Zolpidem dependency and withdrawal seizure: a case report study.

INTRODUCTION: Zolpidem is a short acting inducer of sleep and thought to lack benzodiazepine properties such as anxiolysis, anticonvulsion, muscle relaxation and side effects such as dependency. Recently, some cases of Zolpidem abuse and dependency have been reported. In review of literature, we found that the lowest reported dosage of Zolpidem, which caused dependency, was 160 mg daily. CASE PRESENTATION: We reported a 30-year-old unmarried Iranian woman with dysthymic disorder and chronic insomnia treated with Zolpidem irregularly. She started to use Zolpidem with 5mg per day irregularly since a year ago but augmented its daily dosage gradually to 100 to 150 mg per day in divided doses. After a period of 16 hours without taking Zolpidem she developed a withdrawal syndrome, with generalized tonic-clonic seizures for two times. She was managed with supportive care and recovered completely. CONCLUSIONS: Zolpidem dependency and withdrawal seizure can occur with a dosage under last reported doses. Therefore, possibility of mentioned problems cannot be excluded at any dosage and physicians should pay more attention to potential of Zolpidem to create these adverse effects.