TMT-based proteomics analysis reveals the protective effect of Xuefu Zhuyu decoction on traumatic brain injury.

Abstract Ethnopharmacological relevance Xuefu Zhuyu decoction (XFZYD) is a traditional Chinese herbal prescription. It is effective in treating traumatic brain injury (TBI). However, the underlying molecular mechanisms remain unclear. Aim of the study This study aimed to reveal the possible mechanisms of XFZYD in treating acute TBI through proteomic clues. Materials and methods Controlled Cortical Impact (CCI) rats were gavaged with XFZYD (9 g/kg/d) or distilled water (equal volume) for three days. The Modified Neurological Severity Score (mNSS), brain water content, HE staining, Nissl staining and immunohistochemistry were performed to assess the effects of XFZYD in treating TBI. Additionally, Tandem mass tag-based (TMT) quantitative proteomics technology was applied to detect proteins of the extracted brain cortex. Bioinformatics analysis including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, Protein-protein interaction (PPI) networks were used to analysis differentially expressed proteins (DEPs). Bioinformatics Analysis Tool for Molecular mechanism of TCM (BATMAN-TCM) was applied to anchore diseases and pathways of XFZYD. Besides, western blotting and immunofluorescence were used to verify related proteins. Results XFZYD improved neurologic function, reduced encephaledema and ameliorated cell morphology around the injured area in CCI rats. A total of 6099 proteins were identified with FDR (false discovery rate)  Conclusions Our study suggests that XFZYD may exert therapeutic effects through multi-pathways regulation in the treatment of TBI. This work may provide proteomics clues for the continuation researches on XFZYD in treating TBI.