In vitro and in vivo anti-seizure activity of hydromethanolic extract and fractions of Pterolobium stellatum

Abstract

Ethnopharmacological relevance: In Ethiopia, the whole plant juice of Pterolobium stellatum is used to treat seizures and epilepsy.

Aim of the study: To investigate the antiseizure activity of hydromethanolic crude extract and fractions collected from leaves of P. stellatum using both in vitro, and in vivo seizure models in mice.

Materials and methods: Fresh leaves of P. stellatum were collected from Awash Melka, Addis Ababa, Ethiopia. An 80% crude methanol extract was further fractionated to produce petroleum ether, chloroform, butanol, and aqueous fractions. Anti-seizure activity of the crude extract and fractions (petroleum ether, chloroform, butanol, and water) were assessed at a concentration of 0.7 mg/ml using the in vitro 0 Mg²⁺ model of seizures in mouse brain slices prepared from 14- to 21-day-old C57BL/6 mice. The maximal electroshock seizure (MES) model and the pentylenetetrazol (PTZ) seizure model for seizures were performed on male BALB/c mice using 400 mg/kg and 800 mg/kg of crude 80% methanol extract, as well as the four fractions described above. Diazepam and phenytoin were used as positive controls for PTZ and MES test respectively.

Results: Addition of 0.7 mg/ml of crude 80% methanol extract of P. stellatum prevented the onset of SLEs in most brain slices in the 0 Mg²⁺in vitro model of seizures, with similar efficacy to diazepam (3 μM). The same extract at 400 and 800 mg/kg was efficacious in reducing the hindlimb extension time in the MES model and delaying the onset of myoclonic convulsions in the PTZ model, although not to the same extent as phenytoin (10 mg/kg) or diazepam (5 mg/kg). The chloroform and water fractions of the crude extract also showed significant anti-seizure activity across all three models whilst the non-polar petroleum ether and butanol fractions did not. The UPLC-MS analysis indicated the presence of gallic acid, ellagic acid, kaempferol, myricitrin, isoquercitrin and quercitirin in the crude extract. Gallic acid and ellagic acid were observed in chloroform fraction and in the water fraction ellagic acid, kaempferol, myricitrin and isoquercitrin were detected.

Conclusion: The crude hydromethanolic extract of P. stellatum has significant anti-seizure activity. The chloroform and aqueous fractions have antiseizure activity. The extracts have previously identified compounds with anticonvulsant activity which indicates the antiseizure potential of the plant.

Keywords: 0 Mg(2+) model; Anti-epileptic; Epilepsy; Maximal electroshock seizure model; Pentylenetetrazol seizure model; Pterolobium stellatum; Seizures.