TRIB1 silencing attenuates epilepsy by restoring mitochondrial homeostasis and suppressing microglia-driven neuroinflammation via MAPK pathway inhibition

We aim to explore the role of Tribbles homolog 1 (TRIB1) in epilepsy (EP), specifically its modulation of mitochondrial homeostasis and the mitogen-activated protein kinase (MAPK) pathway. Gene expression profiling, differentially expressed genes screening, functional enrichment analysis, and protein-protein interaction (PPI) network construction were conducted to identify hub genes. Lipopolysaccharide (LPS)-induced BV2 cell models and lithium-pilocarpine-induced EP rat models were established to explore the impact of TRIB1 knockdown on EP development. The severity of EP in rats was evaluated by Racine scale, hematoxylin-eosin staining, and Nissl staining. Cell dysfunction was assessed by detecting cell viability, apoptosis, and oxidative stress markers. Western blot was applied to detect proteins related to mitochondrial function, microglial activation, and the MAPK pathway. We identified 16 hub genes from PPI networks. Among them, TRIB1 was upregulated in EP rats. In EP rat models, TRIB1 knockdown reduced seizure severity, improved oxidative stress, and enhanced mitochondrial homeostasis. TRIB1 knockdown increased viability, inhibited apoptosis, and restored mitochondrial homeostasis in LPS-induced BV2 cells. Moreover, TRIB1 knockdown attenuated microglia activation and neuroinflammation both in vivo and in vitro. TRIB1 knockdown suppressed the MAPK pathway in LPS-induced BV2 cells. The activation of the MAPK pathway reversed the alleviating effect of TRIB1 silencing on LPS-induced BV2 cell and mitochondrial function, as well as microglia-induced neuroinflammation. Knockdown of TRIB1 may provide novel therapeutic strategies for managing EP by restoring mitochondrial homeostasis and inhibiting neuroinflammation driven by microglial activation via the MAPK pathway.