Efficacy of 6-nitrobenzo[d]thiazol-2 Amine Derivative (N3) in Mitigating PTZ-Induced Epileptic Conditions Via Modulation of Inflammatory and Neuroprotective Pathways in-vivo Zebrafish.

Abstract

Epilepsy, a chronic neurological illness affecting 50 million people worldwide, causes recurring seizures due to abnormal brain activity. Current anti-epileptic medications have serious side effects and low efficacy, requiring alternative treatments. The current study investigated the anti-inflammatory, antioxidant, and neuroprotective effects of 6-nitrobenzo[d]thiazol-2-amine (N3) derivatives in a zebrafish larvae model of epilepsy caused by 6 mM pentylenetetrazole (PTZ). Furthermore, N3 was tested for its safety, potential to reduce oxidative stress, inflammation, and neurodegeneration, and effects on motor coordination and neurotransmitter levels. The study utilized in vitro hemolysis assays to evaluate the membrane-stabilizing properties of N3. Zebrafish larvae were pre-treated with N3 at varying concentrations and subsequently exposed to PTZ to induce epilepsy-like conditions. Antioxidant enzyme activities superoxide dismutase (SOD), catalase CAT), glutathione (GSH) levels, lactate dehydrogenase (LDH) activity, and reactive oxygen species (ROS) levels were analyzed. Gene expression for pro-inflammatory and neuroprotective markers was quantified using qPCR, while histological assessments were performed to evaluate amyloid plaque formation, collagen accumulation, and calcium deposition. Behavioral tests measured motor coordination, and gamma-aminobutyric acid (GABA) levels were quantified using high-performance liquid chromatography. N3 demonstrated dose-dependent hemolysis inhibition, confirming its membrane-stabilizing and anti-inflammatory properties up to 43.47 ± 1.36%. It enhanced antioxidant enzyme activities, increased GSH levels 0.76 ± 0.03 nmol/mg, reduced LDH and ROS levels 7.47 ± 0.07 U/mg protein, and suppressed pro-inflammatory gene expression. Histological analysis revealed reduced neurodegenerative markers, including amyloid plaques and calcium deposition. Behavioral improvements were observed, including enhanced motor coordination and increased GABA levels. The findings suggest that N3 derivatives have significant therapeutic potential in epilepsy by reducing oxidative stress, inflammation, and neurodegeneration. Further studies are needed to optimize dosing and confirm safety for clinical applications.