Evaluation of the Antiinflammatory Activity of Dipotassium Glycyrrhizinate in a Cellular Model of Penicillin-induced Epilepsy

• Grant number: 25/11010-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2025
• End date: August 31, 2026
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Manoela Marques Ortega
• Grantee: Gabrielle Pinheiro Trevisan
• Host Institution: Universidade São Francisco (USF). Campus Bragança Paulista. Bragança Paulista , SP, Brazil

Abstract

Introduction: Epilepsy is one of the most prevalent neurological conditions worldwide, affecting over 50 million people and characterized by abnormal and recurrent electrical discharges in the brain. Among its various forms, temporal lobe focal epilepsy stands out as the most common and is frequently refractory to treatment, accounting for a significant portion of drug-resistant cases. The nuclear factor kappa B (NF-¿B) signaling pathway has emerged as a relevant therapeutic target, as its activation is strongly associated with the inflammatory processes triggered by seizures, even in the presence of antiepileptic drugs (AEDs). Glycyrrhizic acid (GA), a bioactive compound extracted from the root of Glycyrrhiza glabra (licorice), exhibits recognized neuroprotective and anti-inflammatory properties, including the ability to inhibit NF-¿B pathway activation in in vivo models. However, its clinical use is limited due to associated hepatic and mineralocorticoid toxicity. Dipotassium glycyrrhizinate (DPG), a salt derived from GA, retains similar pharmacological properties with a more favorable toxicological profile. Nevertheless, its effects on neuroinflammatory processes have not been sufficiently investigated in cellular models that simulate epileptic conditions, such as neuronal hyperexcitability and associated inflammation. Objectives: To evaluate the anti-inflammatory effects of DPG in an in vitro epilepsy model induced by penicillin using the SH-SY5Y cell line. Methods: Initially, an MTT cell viability assay will be performed to determine the subcytotoxic concentration of penicillin-G. Once confluence is reached, SH-SY5Y cells will be exposed to penicillin-G for 30 minutes and 1 hour to induce a neuroinflammatory state similar to that observed in experimental epilepsy models. Control cells will be maintained without exposure to penicillin-G. Subsequently, penicillin-G-exposed cells will be treated with different concentrations of DPG for 24, 48, and 72 hours. The expression of inflammatory microRNAs (miR-146a and miR-155) will be assessed by real-time PCR (qPCR), along with the expression of their NF-¿B pathway-associated target genes (TRAF6 and TNF-¿), as well as pro-inflammatory cytokines such as TNF-¿, IL-1¿, and IL-6. Expected Results: It is expected to identify the lowest effective concentration of DPG capable of preserving cell viability and inhibiting the inflammatory response induced by penicillin-G. This inhibition will be evaluated based on the positive modulation of inflammatory miRNAs (miR-146a and miR-155), associated with repression of NF-¿B pathway target genes and the aforementioned pro-inflammatory cytokines. The findings of this study may provide relevant insights for the development of complementary and safer therapeutic approaches for epilepsy, focusing on neuroinflammation modulation. (AU)