Ramamurthy Karthikeyan, Nayak S P Ramya Ranjan, Madesh S, Panda Siva Prasad, Manikandan K, Rajagopal Rajakrishnan, Alfarhan Ahmed, Palaniappan Senthilkumar, Guru Ajay, Kathiravan M K, Arockiaraj Jesu
Toxicology and Pharmacology Laboratory, Department of Biotechnology, Faculty of Science and Humanities, SRM Institute of Science and Technology, Kattankulathur, Chengalpattu District, Tamil Nadu, 603203, India.
Institute of Pharmaceutical Research, GLA University, Mathura, Uttarpradesh, India.
J Neuroimmune Pharmacol. 2025 Jun 6;20(1):64. doi: 10.1007/s11481-025-10221-6.
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.
癫痫是一种影响全球5000万人的慢性神经疾病,由于大脑活动异常导致反复发作的癫痫。目前的抗癫痫药物有严重的副作用且疗效不佳,需要替代治疗方法。当前的研究在由6 mM戊四氮(PTZ)引起的癫痫斑马鱼幼虫模型中,研究了6-硝基苯并[d]噻唑-2-胺(N3)衍生物的抗炎、抗氧化和神经保护作用。此外,还测试了N3的安全性、降低氧化应激、炎症和神经退行性变的潜力,以及对运动协调和神经递质水平的影响。该研究利用体外溶血试验来评估N3的膜稳定特性。用不同浓度的N3预处理斑马鱼幼虫,随后使其暴露于PTZ以诱导癫痫样状态。分析了抗氧化酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性、谷胱甘肽(GSH)水平、乳酸脱氢酶(LDH)活性和活性氧(ROS)水平。使用qPCR定量促炎和神经保护标志物的基因表达,同时进行组织学评估以评估淀粉样斑块形成、胶原蛋白积累和钙沉积。行为测试测量运动协调性,并使用高效液相色谱法定量γ-氨基丁酸(GABA)水平。N3表现出剂量依赖性的溶血抑制作用,证实其膜稳定和抗炎特性高达43.47±1.36%。它增强了抗氧化酶活性,使GSH水平增加0.76±0.03 nmol/mg,降低了LDH和ROS水平至7.47±0.07 U/mg蛋白质,并抑制了促炎基因表达。组织学分析显示神经退行性标志物减少,包括淀粉样斑块和钙沉积。观察到行为改善,包括运动协调性增强和GABA水平增加。研究结果表明,N3衍生物通过降低氧化应激、炎症和神经退行性变,在癫痫治疗中具有显著的治疗潜力。需要进一步研究以优化给药剂量并确认其临床应用的安全性。
Arch Razi Inst. 2024-10-31