Neuronal nitric oxide synthase/reactive oxygen species pathway is involved in apoptosis and pyroptosis in epilepsy

• Published in: Neural regeneration research Vol. 18; no. 6; pp. 1277 - 1285
• Main Authors: Xu, Xiao-Xue, Shi, Rui-Xue, Fu, Yu, Wang, Jia-Lu, Tong, Xin, Zhang, Shi-Qi, Wang, Na, Li, Mei-Xuan, Tong, Yu, Wang, Wei, He, Miao, Liu, Bing-Yang, Chen, Gui-Lan, Guo, Feng
• Format: Journal Article
• Language: English
• Published: India Wolters Kluwer India Pvt. Ltd 01.06.2023; Medknow Publications & Media Pvt. Ltd; Department of Neurology,the First Affiliated Hospital of China Medical University,Shenyang,Liaoning Province,China%Department of Pharmaceutical Toxicology,School of Pharmaceutical Science,China Medical University,Shenyang,Liaoning Province,China%Department of Neurology,the First Affiliated Hospital of China Medical University,Shenyang,Liaoning Province,China%Key Laboratory of Medical Electrophysiology,Ministry of Education&Medical Electrophysiological Key Laboratory of Sichuan Province,Institute of Cardiovascular Research,Southwest Medical University,Luzhou,Sichuan Province,China%Department of Endocrinology and Metabolism,the Fourth Affiliated Hospital of China Medical University,Shenyang,Liaoning Province,China%Department of Endocrinology,Shengjing Hospital of China Medical University,Shenyang,Liaoning Province,China; Department of Pharmaceutical Toxicology,School of Pharmaceutical Science,China Medical University,Shenyang,Liaoning Province,China; Wolters Kluwer - Medknow; Wolters Kluwer Medknow Publications
• Subjects: Apoptosis; apoptosis; bioinformatics analysis; cell death; epilepsy; nitric oxide synthase; oxidative stress; pyroptosis; rna sequencing; tremor rat; weighted gene co-expression network analysis; Cytokines; Dehydrogenases; Epilepsy; Nitric oxide; Oxidative stress; Reactive oxygen species; RNA sequencing; epilepsy; pyroptosis; weighted gene co-expression network analysis; cell death; apoptosis; nitric oxide synthase; bioinformatics analysis; oxidative stress; Tremor rat
• ISSN: 1673-5374; 1876-7958
• DOI: 10.4103/1673-5374.357906

Dysfunction of neuronal nitric oxide synthase contributes to neurotoxicity, which triggers cell death in various neuropathological diseases, including epilepsy. Studies have shown that inhibition of neuronal nitric oxide synthase activity increases the epilepsy threshold, that is, has an anticonvulsant effect. However, the exact role and potential mechanism of neuronal nitric oxide synthase in seizures are still unclear. In this study, we performed RNA sequencing, functional enrichment analysis, and weighted gene coexpression network analysis of the hippocampus of tremor rats, a rat model of genetic epilepsy. We found damaged hippocampal mitochondria and abnormal succinate dehydrogenase level and Na+-K+-ATPase activity. In addition, we used a pilocarpine-induced N2a cell model to mimic epileptic injury. After application of neuronal nitric oxide synthase inhibitor 7-nitroindazole, changes in malondialdehyde, lactate dehydrogenase and superoxide dismutase, which are associated with oxidative stress, were reversed, and the increase in reactive oxygen species level was reversed by 7-nitroindazole or reactive oxygen species inhibitor N-acetylcysteine. Application of 7-nitroindazole or N-acetylcysteine downregulated the expression of caspase-3 and cytochrome c and reversed the apoptosis of epileptic cells. Furthermore, 7-nitroindazole or N-acetylcysteine downregulated the abnormally high expression of NLRP3, gasdermin-D, interleukin-1β and interleukin-18. This indicated that 7-nitroindazole and N-acetylcysteine each reversed epileptic cell death. Taken together, our findings suggest that the neuronal nitric oxide synthase/reactive oxygen species pathway is involved in pyroptosis of epileptic cells, and inhibiting neuronal nitric oxide synthase activity or its induced oxidative stress may play a neuroprotective role in epilepsy.