TAK-242 alleviates neuroinflammation and poststroke cognitive impairment by targeting the TLR4/SYK/NF-κB pathway in a cerebral ischemia/reperfusion rat model.

Neuroinflammation plays an essential role in neurological dysfunction in cerebral ischemia and reperfusion injury. Understanding the molecular network associated with neuroinflammation will provide potential targets for the neuroprotective treatment of ischemic stroke. An ischemia/reperfusion (I/R) rat model was established by middle cerebral artery occlusion (MCAO) and verified by laser speckle imaging and neurological function scoring. The ipsilateral cortex was analyzed using transcriptomic sequencing. The resulting transcriptomic dataset was combined with the ischemic stroke-associated C0948008 dataset from DisGeNet to screen differentially expressed genes (DEGs). Protein‒protein interaction (PPI) analysis was performed on DEGs associated with the inflammatory response. The candidate genes were verified using Western blotting or ELISA. I/R rats were subsequently treated with the TLR4 inhibitor TAK-242 to investigate the role of the TLR4/SYK/NF-κB pathway in neuroinflammation during ischemic stroke. A total of 264 genes were identified, most of which were enriched in inflammatory response pathways. PPI analysis of the DEGs associated with the inflammatory response revealed a total of three mutually interacting gene clusters, with TLR4, SYK, and NF-κB serving as the hub genes. The expression of proteins in the TLR4/SYK/NF-κB pathway and inflammatory cytokines significantly upregulated the I/R rats, whereas the trends were dramatically reversed by TAK-242. In addition, TAK-242 efficiently decreased the infarct area, improved neurological function, alleviated poststroke cognitive impairment (PSCI), recovered neuronal viability, and inhibited the overactivation of microglia and astrocytes. These data suggest that the TLR4/SYK/NF-κB pathway plays an important role in neuroinflammation and provide potential targets for neuroprotective therapy for ischemic stroke.