Enhancement of the anticonvulsant effect of the h3 histamine receptor blocker thioperamid against the use of pioglitazone

The purpose of the study was to study the dynamics of foci of epileptic activity, which were reproduced in the cerebral cortex of kindled rats using the sodium salt of benzylpenicillin, under the conditions of individual and combined use of pioglitazone and thioperamid. Under conditions of acute observation in rats kindled with pentylenetetrazole repeated administration (30-35 mg/kg, i.p.), the influence of thioperamid (2,5 and 15,0 mg/kg, i.p.) and pioglitazone(50,0 and 200,0 mg/kg, i.p.) administrations upon penicillin-induced foci in the frontal cortex have been investigated. The data obtained showed that pioglitazone use in a previously ineffective dose (50.0 mg/kg, i.p.) but against the background of the administration of thioperamid (2.5 mg/kg, i.p.) was accompanied by the development of an anticonvulsant effect, which was expressed as inhibition of the amplitude and frequency of epileptic seizures potentials, as well as in reducing the life span of epileptic foci. A similar result indicates the possibility of restoration of sensitivity to the anticonvulsant effects of the studied drugs when they are used together in kindling provoked resistance to the action of antiepileptic drugs. Thus, the development of the antiseizure effect of thioperamid and pioglitazone, which were used in initially not effective dosages (2,5 and 50,0 mg/kg correspondently), have been described. This fact favors the synergy of the antiepileptic action of investigated compounds

Immunohistochemical neuroinflammatory markers in the hippocampus of PTZ-kindled rats under conditions of rapamycin and axitinib treatment

The aim of the study is to determine the level of HIF-1α, TNF-α, and NF-kB in the hippocampus of kindled rats treated with rapamycin and axitinib. Materials and methods. Kindling was produced in 29 rats by administration of three-week pentylenetetrazole (PTZ, 35.0 mg/kg, i.p.). Treatment with rapamycin (0.5 mg/kg, i.p.) and axitinib (2.5 mg/kg, i.p.) was performed for ten days in fully kindled rats. The avidin-biotin-peroxidase method was used for hippocampal slice staining. For negative control, staining was performed using only secondary antibodies. Results. The HIF-1α expression increased in kindled rats raised by 1.77 times compared to the control (p<0.001). Axitinib treatment resulted in of HIF-1α level of 16.7 % (p<0.05) compared with kindled animals, while combined treatment with rapamycin and axitinib reduced HIF-1α by 33.8 % (p<0.01). In kindled rats, TNF-α expression was 3.74 times greater than in control (p<0.001). Rapamycin treatment reduced TNF-α by 31.0 % (p<0.01). Axitinib treatment caused a reduction of TNF-α by 21.1 % (p<0.05). Combined treatment with rapamycin and axitinib reduced TNF-α by 48.0 % (p<0.001) but still exceeded the TNF-α in control by 1.95 times (p<0.01). NF-kB level in kindled rats exceeded the control by three times (p0.05), while axitinib – by 26.5 % (p<0.05) compared with kindled rats. Combined treatment with rapamycin and axitinib resulted in NF-kB reduction by 56.7 % compared with kindled rats (p<0.001). Conclusions. PTZ-kindling resulted in an increase in the immunoreactivity of HIF-1α, TNF-α, and NF-kB in the hippocampus. Combined treatment with rapamycin and axitinib engendered prevention of generalized seizures and normalized the level of HIF-1α and NF-kB with a significant reduction of TNF-α. Effects of treatment favours of synergy action of rapamycin and axitini

ВИВЧЕННЯ ГОСТРОЇ ТОКСИЧНОСТІ ТА ПРОТИЗАПАЛЬНОЇ АКТИВНОСТІ СПИРТОВИХ ЕКСТРАКТІВ ТРАВИ СНУ БІЛОГО (PULSATILA ALBA)

The aim of the work. Research of the acute toxicity and anti-inflammatory (anti-exudative) activity studying of Pulsatila alba ethanolic extracts. Materials and Methods. Acute toxicity studies were performed on 60 white rats of both sexes weighing 190–220 g, which were administered intragastric single-dose of tested extracts at doses of 5000, 10,000 and 15,000 mg/kg. The carragenine model of inflammatory oedema of white rats paw was used for anti-exudative activity studying. The tested extracts were administered intragastrically at a dose of 2 ml/kg. Diclofenac sodium and ketanov were used as reference drugs. Results and Discussion. The results of Pulsatila alba ethanolic extracts acute toxicity studying demonstrated absence of any toxic manifestations in intragastric administration at doses of 5000, 10,000 and 15,000 mg/kg in white rats, which allows their to VI class of toxicity (relatively harmless substances). The Pulsatila alba ethanolic extracts posses a tendency for anti-inflammatory activity in the carrageenan model of inflammatory edema in white rats and the pharmacological effect depends on the concentration of ethanol and the ratio of raw material: extractant. The best anti-exudative activity was detected for 70 % ethanolic extracts (1:20), an inhibition index of inflammatory response was 27 %, which is slightly inferior to reference drugs (by 10 and 17 % relative ketanov and sodium diclofenac, respectively) and is a good level for anti-inflammatory phitopreparations. Conclusions. The acute toxicity and anti-inflammatory effect of Pulsatila alba ethanolic extracts were investigated and it was established that 70 % alcoholic extract (1:20) is a relatively harmless substance (grade VI toxicity) and has potency anti-inflammatory activity (inhibition of inflammation - 27 %) on the carrageenan model of inflammatory edema of the white rat paws at intragastric administration.Мета роботи. Дослідження гострої токсичності та протизапальної (антиексудативної) активності спиртових екстрактів трави сну білого (Pulsatila alba). Матеріали і методи. Дослідження гострої токсичності проведено на 78 безпородних білих щурах обох статей масою 190–220 г, яким вводили тестовані спиртові екстракти трави сну білого (Pulsatila alba) одноразово внутрішньошлунково за допомогою металевого зонда в дозах 5000, 10 000 та 15 000 мг/кг. Для вивчення антиексудативної активності використовували карагенінову модель запального набряку задньої лапи білих щурів (42 тварини). Досліджувані екстракти вводили внутрішньошлунково в дозі 2 мл/кг. Як препарати порівняння використовували диклофенак натрію та кеторолаку трометаміну. Результати й обговорення. Результати вивчення гострої токсичності екстрактів трави сну білого (Pulsatila alba) свідчать про відсутність будь-яких токсичних проявів при внутрішньошлунковому введенні в дозах 5000, 10 000 та 15 000 мг/кг на білих щурах, що дозволяє віднести їх до VI класу токсичності (відносно нешкідливі речовини). Спиртові екстракти трави сну білого проявляють протизапальну активність на карагеніновій моделі запального набряку лапи білого щура і фармакологічний ефект залежить від концентрації етанолу та співвідношення сировина:екстрагент. Найкращу антиексудативну активність проявляє 70 % спиртовий екстракт (1:20), показник пригнічення запальної реакції якого становив 27 %, що дещо поступається референс-препаратам (нижче на 10 та 17 % відносно кеторолаку трометаміну та диклофенаку натрію, відповідно) та є хорошим показником для рослинних протизапальних засобів. Висновки. Досліджено гостру токсичність та протизапальну дію спиртових екстрактів трави сну білого (Pulsatila alba) і встановлено, що 70 % спиртовий екстракт (1:20) є відносно нешкідливою речовиною (VI клас токсичності) та проявляє виразну протизапальну активність (показник пригнічення запальної реакції - 27 %) на карагеніновій моделі запального набряку лапи білого щура при внутрішньошлунковому введенні

Axitinib displays antiseizure activity on pentylenetetrazol – induced kindling mode

This work aimed to investigate the effects of a specific inhibitor of vascular endothelial growth factor receptor generic axitinib upon pentylenetetrazol (PTZ) - induced kindled and acute convulsions and to compare axitinib effects with diazepam. Wistar rats kindled with PTZ (30.0 mg/kg, intraperitoneally) during three weeks up to a stage of generalized convulsions stage 5 were observed. The assessment of seizures was performed immediately after completing the kindling procedure, after PTZ-free two weeks, and on acute PTZ-induced convulsions. Axitinib was administered perorally with PTZ injection 60 min later. Diazepam was administered intraperitoneally followed with PTZ in 30 min. Axitinib prevented generalized convulsions in 5 out of 8 kindled rats in a dose of 2.5 mg/kg, and the pronouncement of effect was comparable with such one caused by diazepam in a dose of 0.1 mg/kg. Axitinib in the dose of 10.0 mg/kg completely protected generalized convulsions. A significant reduction of seizure severity was observed when axitinib (2.5 and 10.0 mg/kg) was administered to kindled rats after two weeks free from PTZ, while diazepam caused the suppression of convulsions only in the highest dose of 1.0 mg/kg. Acute PTZ seizures were suppressed by diazepam in both doses, while axitinib was not effective. Axitinib reduced the density of neomicrovessels in the frontal cortex by 47.0% in early kindling and 43.9% in the postponed period. The effectiveness of axitinib on the kindling model of epilepsy might be explained by preventing neoangiogenesis as a valid mechanism of chronic brain epileptization development

Oxidative stress suppression contributes to antiseizure action of axitinib and rapamycin in pentylenetetrazol-induced kindling

Rapamycin and axitinib block different kinases in signaling pathways such as PI3K-Akt-mTOR and BDNF-TrkB, respectively. Both have antiseizure and antioxidative actions, which justify studying the combined effects of these drugs upon seizures and oxidative stress in the chronic model of epilepsy. The investigation aimed to look for the combined effect of rapamycin and axitinib upon pentylenetetrazol (PTZ)-kindled seizures and oxidative stress. Experiments were performed on 300 two- to four-month-old Wistar male rats, which had been kindled daily with PTZ (35.0 mg/kg, i.p.). Malondialdehyde (MDA) level, superoxide dismutase (SOD) activity, and glutathione (GSH) level were determined in brain tissues of kindled rats before and after the treatment. The analysis of antiseizure and antioxidative actions was performed using ED50 of rapamycin and axitinib, with their combined administration using graded dosages of ED50 of each drug. The median effective dose (ED50 ) for rapamycin and axitinib was 0.93 and 4.97 mg/kg, respectively. ED50 of rapamycin when combined with axitinib (2.0 mg/kg) was 0.60 mg/kg, which was reduced by 35.6% when compared with the ed 50 administered alone (P < 0.05). The MDA level increased from 152.9±24.8 to 388.3±49.2 nmol/mg of protein (P < 0.05), while SOD activity reduced from 11.14±2.33 to 3.54±1.08 IU/mg of protein (P < 0.05) in brain tissues of the kindled rats. Combined treatment with rapamycin (0.56 mg/kg, i.p.) and axitinib (2.0 mg/ kg, i.p.) resulted in a significant rise in SOD activity (11.09±1.86 IU/mg) and GSH level (7.32±1.34 µg/mg) when compared with the kindled rats (P < 0.05). Combined axitinib and rapamycin therapy have an antiepileptic and antioxidative effect on PTZ-kindled seizures

Blockade of Н3 histamine receptors facilitates antiseizure action of Pentoxyphilline

Considering the important role of inflammation in epilepsy development is reasonable to investigate the effects of combined usage of different modulators of neural inflammation. The work aimed to investigate the effects of the blocker of histamine Н3 pitolisant hydrochloride (Selleck, USA) and pentoxyphilline (Sigma-Aldrich, USA), which prevents proinflammatory cytokines elaboration upon pentylenetetrazol (Sigma-Aldrich, USA) (PTZ)-kindled seizures in rats. Male Wistar rats four months of age kindled to the stage of generalized tonic-clonic seizure fits with PTZ (35 mg/kg, i.p.) were used in observations. Pentoxyphilline (50 mg/kg, i.p.) administered with pitolisant (5 mg/kg, i.p.) caused the increase of first seizure manifestations latency by 67.2% in comparison with the control group of kindled rats (P<0.05)

Enhancement of the anticonvulsant effect of the h3 histamine receptor blocker thioperamid against the use of pioglitazone

The purpose of the study was to study the dynamics of foci of epileptic activity, which were reproduced in the cerebral cortex of kindled rats using the sodium salt of benzylpenicillin, under the conditions of individual and combined use of pioglitazone and thioperamid. Under conditions of acute observation in rats kindled with pentylenetetrazole repeated administration (30-35 mg/kg, i.p.), the influence of thioperamid (2,5 and 15,0 mg/kg, i.p.) and pioglitazone(50,0 and 200,0 mg/kg, i.p.) administrations upon penicillin-induced foci in the frontal cortex have been investigated. The data obtained showed that pioglitazone use in a previously ineffective dose (50.0 mg/kg, i.p.) but against the background of the administration of thioperamid (2.5 mg/kg, i.p.) was accompanied by the development of an anticonvulsant effect, which was expressed as inhibition of the amplitude and frequency of epileptic seizures potentials, as well as in reducing the life span of epileptic foci. A similar result indicates the possibility of restoration of sensitivity to the anticonvulsant effects of the studied drugs when they are used together in kindling provoked resistance to the action of antiepileptic drugs. Thus, the development of the antiseizure effect of thioperamid and pioglitazone, which were used in initially not effective dosages (2,5 and 50,0 mg/kg correspondently), have been described. This fact favors the synergy of the antiepileptic action of investigated compounds

Prevention of formalin-induced nociceptive behaviors caused by ppar-gamma activation strengthened with cerebellar transcranial direct current stimulation

The investigation was performed on 55 male Wistar rats. The pain behaviors were induced through the formalin (2.5%) subcutaneous administration into the plantar surface of the right hind paw. Pain score value was determined continuously during 90 min. It was established that combined usage of pioglitazone (100.0 mg/kg, i.p.) and cerebellar transcranial direct current stimulation (tDCS) (300 µA, 10.0 min) resulted in pronounced protection of pain behaviors at all stages of the pain syndrome development. The severity of investigated index decreased by 60.0% compared to the control data at 4-6 min after formalin administration (P<0.05). Averaged severity of pain behaviors was suppressed more than twice at Phase I and Interphase (P<0.05), by 60.0% at Phase 2A (P<0.05), and by 33.3% at Phase 2B (P<0.05). Pain -suppressive effect encompassed 42.0+8.25 min of Phase 2A and exceeded data in the group treated with tDCS by 39.1% (P<0.05) and in the group treated with pioglitazone by 64.3% (P<0.05). The conclusion was made that the pain-protective effect of pioglitazone is strengthened with cerebellar tDCS

Effects of a Combination of Transcranial DC Cerebellar Stimulation and Pioglitazone Administration on Pentylenetetrazole-Induced Seizures in Kindled Rats

In rats with the kindling syndrome induced by i.p. injections of pentylenetetrazole (PTZ; 35 mg/kg daily, for three weeks), the averaged latency of seizures induced by a test injection of PTZ (35 mg/kg) was significantly greater (by 42.2%, P < 0.05 vs. control) after a combination of pioglitazone administration (100 mg/kg, i.p.) and transcranial DC stimulation (tDCS, 300 μA, 10 min, cathode on the skull surface) oriented toward the cerebellar cortex. Also, combined using of tDCS and pioglitazone prevented the occurrence of generalized tonico-clonic seizure attacks in 8 out of 10 rats of the respective group (P < 0.05), reduced the seizure severity by 31.3% on average (P < 0.05), and made shorter ictal discharges (by 45.0%; P < 0.05)

Pentylenetetrazole-induced kindling as a model metabolic syndrome

Data on the relationship between epilepsy and metabolic syndrome justified the evaluation of MS markers expression in PTZ-kindled rats. PTZ kindling was induced in four months aged male Wistar rats with three weeks epileptogen (35.0 mg/kg, i.p.) administration. Those animals with fully developed generalized seizures were used for investigations. The glucose tolerance test (GTT) was followed by a significant rising in blood glucose level 30 min after glucose i.p. administration (2.0 g/kg) up to 395.7+47.2 mg/dl), which exceeded the control data by 1.74 times (P<0.001). Insulin tolerance test (0.75 U of insulin/kg) revealed in 15 min after glucose i.p. administration (2.0 g/kg) up to 114.3+15.7 mg/dl), which exceed the control data by 31.3% (P<0.001)