Screening of liothyronine network pharmacology role in the treatment of ischemic stroke and molecular mechanism

Abstract

AbstractObjectiveThe present study aimed to elucidate mechanisms of liothyronine on the treatment of ischemic stroke (IS).MethodsDifferential analysis based on R limma package was used to identify differentially expressed genes, which were then mapped into the connectivity map database for identification of liothyronine associated with IS. Tumor necrosis factor (TNF) signaling pathway was verified through pathway enrichment analysis via Enrichr online. Ischemia stroke mouse model was built up for further analysis. Infarct area and regional cerebral blood flow (rCBF) were measured by 2, 3, 5‐triphenyltetrazolium chloride and laser Doppler flowmetry, respectively. Light microscope was used for the evaluation of body weight and dark neurons. Serum TXB2, 6‐Keto‐PGF1a, TNF‐α, and interleukin‐6 (IL‐6) levels in mice were measured using enzyme‐linked immuno sorbent assay. In addition, relative protein expression levels of brain‐derived neurotrophic factor, nestin, and Sox2 were detected by Western blot analysis.ResultsLiothyronine with a negative connectivity was identified as one promising treatment for IS through TNF signaling pathway. The experimental results showed that liothyronine treatment significantly meliorated infarct area and the number of dark neurons in IS mice. Liothyronine greatly ameliorated the expression levels of TXB2 and 6‐Keto‐PGF1a. Besides, rCBF and body weight change of IS mice were increased gradually with increase of drug concentration. Based on pathway enrichment analysis, anti‐inflammatory response (TNF‐α and IL‐6) relevant to TNF signaling pathway was identified, which was further validated in vitro. Furthermore, proteins as neural stem cell markers made a difference with liothyronine treatment.ConclusionLiothyronine may be a novel therapeutic component to exploit an effective medicine for the treatment of IS.