Beneficial Effects of Rosmarinic Acid In Vitro and In Vivo Models of Epileptiform Activity Induced by Pilocarpine

Abstract

Epilepsy is characterized by a predisposition to generate recurrent and spontaneous seizures; it affects millions of people worldwide. Status epilepticus (SE) is a severe type of seizure. In this context, screening potential treatments is very important. In the present study, we evaluated the beneficial effects of rosmarinic acid (RA) in pilocarpine-induced in vitro and in vivo models of epileptiform activity. Using an in vitro model in combined entorhinal cortex–hippocampal from Wistar rats we evaluated the effects of RA (10 µg/mL) on the lactate release and a glucose fluorescent analogue, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NDBG), after incubation in high potassium aCSF supplemented or not with pilocarpine. In the in vivo model, SE was induced in male C57BL/6 mice by pilocarpine. At 1, 24, and 48 h after the end of SE mice were treated with RA (30 mg/kg/v.o.). We evaluated the neuromotor impairment by neuroscore tests and protein carbonyl levels in the cerebral cortex. In both in vitro models, RA was able to decrease the stimulated lactate release, while no effect on 2-NBDG uptake was found. RA has beneficial effects in models of epileptiform activity in vivo and in vitro. We found that RA treatment attenuated SE-induced neuromotor impairment at the 48 h timepoint. Moreover, post-SE treatment with RA decreased levels of protein carbonyls in the cerebral cortex of mice when compared to their vehicle-treated counterparts. Importantly, RA was effective in a model of SE which is relevant for the human condition. The present data add to the literature on the biological effects of RA, which could be a good candidate for add-on therapy in epilepsy.

Effects of Rosmarinus officinalis L. Extract on Neurobehavioral and Neurobiological Changes in Male Rats with Pentylenetetrazol-Induced Epilepsy

“…In response to lipopolysaccharide, SOD 3 was released from intracellular stores of macrophages producing biological responses [49]. Lactate has been considered a useful seizure-like activity biomarker, as demonstrated by an increase in its levels in the extracellular environment of the human hippocampus during seizures; rosmarinic acid (30 mg/kg/body volume) was able to reduce pilocarpine-stimulated lactate release in the brains of epileptic male rats [50]. Gap junctions that allow direct transfer of ions and molecules between neighboring cells, and gap junctions between astrocytes, play an important role in the development of brain diseases such as epilepsy [51].…”

Comprehensive Insights into Biological Roles of Rosmarinic Acid: Implications in Diabetes, Cancer and Neurodegenerative Diseases

“…A study investigated the potential advantages of RA in both in vitro and in vivo models of epileptiform activity induced by pilocarpine. This study revealed RA's potential to reduce the acute neuromotor disturbances and oxidative damage triggered by status epilepticus (SE) in mice [68]. Furthermore, it appears to offer favourable effects in in vitro models of epileptiform activity, as evidenced by a decrease in lactate release.…”

“…RA doses of 100 µg/mL significantly inhibited the activity of gammaaminobutyric acid transaminase (GABA-T) [67]. Numerous reports showed the protective role of RA in animal models and in vitro studies against epilepsy [68,69]. A study investigated the potential advantages of RA in both in vitro and in vivo models of epileptiform activity induced by pilocarpine.…”