Seven week old adult male Sprague-Dawley (SD) rats (Koatech, Seoul, Korea) were used, and maintained at standard room temperature (22–24 °C) with 12 hour light and dark cycle. Rats were infused with lithium chloride (127 mg/kg, Sigma-Aldrich, MO, USA) and methylscopolamine-bromide (1 mg/kg, Sigma-Aldrich, MO, USA) via intraperitoneal route 24 h and 30 min prior to pilocarpine administration, respectively. Pilocarpine hydrochloride (30 mg/kg, i.p., Sigma-Aldrich, MO, USA), a muscarinic cholinergic receptor agonist was injected to trigger status epilepticus (SE). The seizure events were scored in pilocarpine-induced rats through repeated doses of pilocarpine hydrochloride (10 mg/kg) every 30 min according to the Racine scale41. Racine scale was defined as adequate measurement of seizure intensity with a 5-stage spontaneous seizure scale. Diazepam (10 mg/kg, i.p. Samjin, Seoul, Korea) was injected 60 min after the onset of seizure to reduce seizure activity. After cessation of SE, rats were given 5 ml intraperitoneal injection of 0.9% saline for hydration and treated with sufficient moistened pellets soaked in Gatorade. Control rats received lithium chloride, methylscopolamine-bromide and the same amount of 0.9% saline instead of pilocarpine.
