components was achieved by comparison of their retention times and UV spectra Riluzole relative to standards chromatographed under the same conditions. In addition, confirmation of the identified compounds was performed by analysing standard and sample solutions using an HPLC MS equipped with an ESI interface in negative ionisation mode under the same chromatographic conditions. Drugs. Pentylenetetrazole, hexobarbital, naloxone hydrochloride dihydrate, atropine sulfate salt monohydrate and flumazenil were purchased from Sigma Aldrich Chemical Company, St Louis, MO,USA.Diazepam experiments were performed with a commercially available DiazemW ampoule. Animals. Adult male Swiss albino mice weighing 35 40 g were used for the experiments.
Animals were maintained in a 12 h light/dark cycle and were acclimatized to the laboratory environment for at least 48 h before testing. The experimental protocols were approved by the Local Ethical Committee on Animal Experimentation of Eskiehir Anadolu University, Turkey. Administration of the extract. The animals were divided randomly GSK-3 alpha inhibitor into the following groups: control group, reference group, extract treated test groups, and antagonist pre treated plus extract treated test groups. The control solution was physiological saline. Physiological saline and different doses of the extract were orally administered 60min before the experimental sessions. Mechanistic studies were conducted using Flu, a g aminobutyric acid A benzodiazepine receptor complex site antagonist, Nlx, a non specific opioid receptor antagonist, and Atr, a non selective muscarinic receptor antagonist.
The reference drug Dzm was used at 2mg/kg in the hole board, activity cage, PTZ induced seizure and Hxbinduced sleeping tests. Antagonist compounds, Imiquimod 99011-02-6 including Nlx at 5mg/kg, Atr at 2mg/kg and Flu at 2.5 and 10mg/kg, were administered intraperitoneally 15min before administration of the extract. Assessment of exploratory behaviour in hole board tests. The exploratory behaviour of mice was examined using a hole board apparatus, as described previously. The hole board apparatus consisted of a grey Perspex panel with 16 equidistant holes of 3 cm diameter in the floor. The board was positioned 15 cm above the ground. Head dips was measured by infrared cells placed under the holes. Each animal was placed individually in the centre of the board facing away from the observer and was allowed to explore the apparatus freely.
At the end of each test, the animal was removed and the floor was cleaned. The total number of head dipping behaviours over 5 min was recorded. Lapatinib 388082-77-7 Assessment of spontaneous locomotor activity in activity cage tests. Spontaneous locomotor activity of the mice was measured using an activity cage apparatus containing two sets of 16 photocells placed 3 cm brain and 12 cm above the floor under a transparent cover. Interruptions of light beams from the photocells were registered during horizontal and vertical movements of the animals. The total number of horizontal and vertical locomotor activities of the mice were recorded for 4 min. Assessment of sedative activity in Hxb induced sleeping tests. To evaluate the presence of sedative activity, a Hxb induced sleeping test was performed. Hxb at a dose of 60 mg/kg was administered to the mice to induce.
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