Evaluation of the anti-inflammatory activity of glycyrrizinate dipotassium in a murine model of epilepsy induced by penicillin

• Grant number: 24/06901-4
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2025
• End date: January 31, 2027
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Manoela Marques Ortega
• Grantee: Gabriel Eugênio Inácio dos Santos
• Host Institution: Universidade São Francisco (USF). Campus Bragança Paulista. Bragança Paulista , SP, Brazil

Abstract

Introduction: Epilepsy is one of the most common neurological disorders, affecting over 50 million people worldwide, characterized by abnormal brain electrical activity causing seizures or altered consciousness. Approximately two-thirds of individuals with epilepsy have temporal lobe epilepsy, the most common type of drug-resistant epilepsy, justifying the search for new treatments, mainly due to the side effects and high prices of current treatments. The nuclear factor kappa B (NF-¿B) signaling pathway is active in the central nervous system in response to inflammation observed after seizures in people with epilepsy, even those using antiepileptic drugs. Glycyrrhizic acid (GA), a compound isolated from licorice root (Glycyrrhiza glabra), has been evaluated in previous studies in in vivo models of epilepsy induction for its anti-inflammatory effects. Thus, GA was able to reduce malondialdehyde (MDA), responsible for post-seizure neural apoptosis, with morphologically normal hippocampal neurons and inhibition of the NF-¿B pathway. However, at doses from 100mg/day, GA showed toxicity. Dipotassium glycyrrhizinate (DPG), a dipotassium salt of glycyrrhizic acid (GA), possesses anti-inflammatory properties without observed in vivo side effects of GA. Furthermore, previous studies from our group observed that DPG inhibited the NF-¿B pathway through the modulation of microRNAs (miRs) miR-16 and miR-146a, which inhibited the expression of their respective target genes IRAK2 and TRAF6, belonging to the NF-¿B pathway, using glioblastoma cell lines (in vitro), a tumor type that constantly activates the pathway. Objectives: Therefore, the aim of this study will be to evaluate the effect of DPG in in vivo models of penicillin-induced epilepsy. Additionally, the inhibition of the NF-¿B pathway by the modulation of miR-146a, miR-155, and miR-194-5p and their predicted target genes belonging to the NF-¿B pathway, TRAF6, TNF-¿, and HMGB, respectively, will be evaluated. Methods: After approval from the Committee on Ethics in Animal Use (CEUA), 64 mice will be divided into 4 groups of 15 animals plus test group (n=4): 1) Control group; 2) Clobazam group; 3) DPG group; 4) Clobazam + DPG group. Firstly, seizure induction in chemically induced animal models by intracortical penicillin injection will be validated by behavioral analysis. Then, the animals will be treated with DPG, and the efficacy of DPG in inhibiting the NF-¿B pathway by the modulation of previously selected miRs and their predicted target genes will be assessed. The miRs and predicted genes will be evaluated by real-time polymerase chain reaction (PCR). Expected results: DPG could be safely used in controlling epileptic seizures, concurrently with synthetic drugs recommended for individuals with drug-responsive epilepsy and/or drug-resistant epilepsy.