Journal Article

NeuroAid™-II (MLC901) provides neuroprotection and enhances neuronal cell survival against kainic acid-induced excitotoxicity in vitro by activating the PI3K/AKT pathway

Yazid, Muhammad Dain, Naicker, Amaramalar Selvi, Daud, Muhammad Fauzi, Lokanathan, Yogeswaran, Ismail, Ohnmar Htwe, Idris, Jalilah, Athi Kumar, Ramesh Kumar, Anjum, Anam, Ng, Angela Min Hwei

2025

Overview

In the twenty-first century, in vitro models of excitotoxic injury, leveraging advanced cellular and molecular technologies, offer a promising alternative to animal studies. These models provide a more precise understanding of injury mechanisms. Method: This study utilizes kainic acid (KA), a potent glutamate receptor agonist, to induce excitotoxicity, oxidative stress, and mitochondrial dysfunction, resulting in motor neuron (MN) degeneration. Mature, differentiated NSC-34 MNs were exposed to different concentrations of KA (0.1, 0.5, and 1 mM) to induce neurodegeneration and apoptosis. Following KA treatment, cells were either treated with MLC901 (NeuroAiD™ II) or left untreated. The effects were assessed through cell viability assays, immunocytochemistry with antibiotic staining, and analysis of key markers in the PI3K/AKT signaling pathway. KA exposure resulted in significant neurodegeneration and apoptosis, as indicated by a reduction in cell viability, a decrease in Tubulin beta-III expression, and downregulation of regenerative markers, including AKT, p-AKT, and GAP43. Additionally, the apoptotic marker p-GSK3β was upregulated in KA-treated cells. In contrast, MLC901 treatment alleviated these detrimental effects. MLC901 restored Tubulin beta-III expression and reversed the downregulation of PI3K/AKT signaling markers (AKT, p-AKT, GAP43). Furthermore, MLC901 treatment led to a reduction in the apoptotic marker p-GSK3β. This study demonstrates that KA induces necrotic and apoptotic cell damage, mimicking secondary injury mechanisms typical of neurodegenerative diseases. MLC901 shows promise as a neuroprotective agent, counteracting KA-induced excitotoxicity and highlighting its potential therapeutic application in neuroprotection.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V,SpringerOpen
• Subject: Antibiotics / Cell culture / Cell death / Chemical injury / Enzymes / Excitotoxicity / In vitro model / Kainic acid / Medicine / Medicine & Public Health / MLC901 / Nervous system / Neuroblastoma / Neurodegeneration / Neurons / Neuroprotection / Neurotoxicity / Oxidative stress / Proteins / Software / Spinal cord / Traumatic brain injury / Variance analysis