Effects of toluene on the protein expression profiles of rat cochlear outer hair cells
Objective To filter the differentially expressed proteins(DEPs)in outer hair cells(OHCs)of rat cochlear after toluene exposure,and analyze the functions and interactions of DEPs.Methods Rat cochlear OHCs were cultured in the 1640 basal medium containing toluene at different doses(concentrations:0,5,10,20,40,50,60,80,and 100 mmol/L).Cell activity was detected by CCK-8,and the changes of protein expression profile of rat cochlear OHCs in the exposed group and the control group were detected by Q Extractive mass spectrometry.DEPs were analyzed by Perseus software and their biological functions were analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.The expression of hub proteins was detected by Western Blot(WB).Results A total of 233 DEPs were screened,including 157 up-regulated proteins and 76 down-regulated proteins.GO analysis results showed that the functions of DEPs were mainly involved in glycolytic,gluconeogenesis and glycoprotein binding processes.KEGG enrichment analysis showed that DEPs were mainly related to dysregulation of glycolysis/gluconeogenesis and extracellular exosome pathways.Three key proteins,namely triosephosphate isomerase 1(TPI1),phosphoglycerate kinase 1(PGK1)and fructose-bisphosphate Aldolase A(ALDOA),were identified through protein-protein interaction(PPI)analysis and screening of proteins involved in the dysregulated pathway of glycolysis/gluconeogenesis.WB results showed that TPI1,PGK1 and ALDOA protein expression levels in rat cochlear OHCs were decreased after toluene exposure.Conclusion Exposure to toluene may cause dysregulation of glucose metabolism pathways and lead to cochlear cell damage in rats by inhibiting the expression of TPI1,PGK1,and ALDOA proteins.
TolueneOuter hair cells of rat cochlearDifferentially expressed proteins
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