Study On The Expression And Regulation Mechanisms Of LncRNAs In Age-Related Hearing Loss In A Mouse Model

Objective:Age-related hearing loss(AHL)is an important health problem in the elderly population and a complex multi factorial disease and its molecular mechanisms have not been fully elucidated.An increasing number of lncRNAs have been reported to be associated with the induction of cell apoptosis,tumorigenesis,and neurological disorders.In this study,we analyzed the important role of lncRNA in the development of mouse neurological disorders AHL through RNA-Seq analysis.Methods:First,we analyzed the differential expression of lncRNAs and mRNAs in the cochleae of six-week-old and one-year-old C57BL/6J mice through RNA-Seq analysis.Gene ontology analysis was performed,and we selected hearing loss,oxidative stress and apoptosis-related differentially expressed lncRNAs by the bioinformatic analysis on hearing loss related gene database(https://hereditaryhearingloss.org/updates).qRT-PCR was performed to validate the expression levels of lncRNAs and their coexpressed mRNAs in animal and cellular models of AHL.A smart silencer targeting lncRNA(lncRNAi)was designed to knock down lncRNA in HEI-OC1 cells.qRT-PCR and Western blotting analysis suggest that the relationship of lncRNA and the expression of oxidative stress and apoptosis-related gene Ar,hearing loss-related gene Hgf and the protein caspase 3 and HO-1.Results:RNA-Seq analysis suggests that we found 738 and 2033 differentially expressed lncRNAs and mRNAs,respectively,in these two groups(corrected P<0.05).There are 29 mRNAs enriched in the sensory perception of sound(GO:0007605).qRT-PCR results suggested among these lncRNAs,lncRNA NONMMUT010961.2 was the most markedly different.qRT-PCR results suggested that Hgf and Ar were downregulated in lncRNA NONMMUT010961.2 knockdown cells compared with NC cells.Western blotting results showed that the expression levels of cleaved caspase 3 and HO-1 were decreased by lncRNAi in the AHL cellular model.Conclusion:Our results suggest that lncRNA and mRNA dysregulations is involved in the development of AHL in C57BL/6J mice.Preliminary results suggested that lncRNAs NONMMUT010961.2 may be associated with AHL by regulating the expression of Ar and Hgf in mice and may thus lead to a new treatment for AHL.