Alleviation of hippocampal necroptosis and neuroinflammation by NecroX-7 treatment after acute seizures

Yihyun Roh; Su Bin Lee; Minseo Kim; Mi-Hye Kim; Hee Jung Kim; Kyung-Ok Cho

doi:10.3389/fphar.2023.1187819

Alleviation of hippocampal necroptosis and neuroinflammation by NecroX-7 treatment after acute seizures

Front Pharmacol. 2023 Aug 2:14:1187819. doi: 10.3389/fphar.2023.1187819. eCollection 2023.

Authors

Yihyun Roh¹, Su Bin Lee², Minseo Kim¹, Mi-Hye Kim², Hee Jung Kim³, Kyung-Ok Cho^{4

5}

Affiliations

¹ College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
² Department of Medical Laser, Graduate School, Dankook University, Cheonan, Republic of Korea.
³ Department of Physiology, College of Medicine, Center for Human Risk Assessment, Dankook University, Cheonan, Republic of Korea.
⁴ Department of Pharmacology, Catholic Neuroscience Institute, Institute for Aging and Metabolic Diseases, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
⁵ Department of Biomedicine and Health Sciences, The Catholic University of Korea, Seoul, Republic of Korea.

Abstract

Temporal lobe epilepsy (TLE) is one of the most common neurological disorders, but still one-third of patients cannot be properly treated by current medication. Thus, we investigated the therapeutic effects of a novel small molecule, NecroX-7, in TLE using both a low [Mg²⁺]_o-induced epileptiform activity model and a mouse model of pilocarpine-induced status epilepticus (SE). NecroX-7 post-treatment enhanced the viability of primary hippocampal neurons exposed to low [Mg²⁺]_o compared to controls in an MTT assay. Application of NecroX-7 after pilocarpine-induced SE also reduced the number of degenerating neurons labelled with Fluoro-Jade B. Immunocytochemistry and immunohistochemistry showed that NecroX-7 post-treatment significantly alleviated ionized calcium-binding adaptor molecule 1 (Iba1) intensity and immunoreactive area, while the attenuation of reactive astrocytosis by glial fibrillary acidic protein (GFAP) staining was observed in cultured hippocampal neurons. However, NecroX-7-mediated morphologic changes of astrocytes were seen in both in vitro and in vivo models of TLE. Finally, western blot analysis demonstrated that NecroX-7 post-treatment after acute seizures could decrease the expression of mixed lineage kinase domain-like pseudokinase (MLKL) and phosphorylated MLKL (p-MLKL), markers for necroptosis. Taken all together, NecroX-7 has potential as a novel medication for TLE with its neuroprotective, anti-inflammatory, and anti-necroptotic effects.

Keywords: NecroX-7; excitotoxicity; hippocampus; necroptosis; neuroinflammation; neuroprotection; seizure.

Grants and funding

This research was funded by MitoImmune Therapeutics Inc. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.