Association And Mechanism Of STAT3 Gene Polymorphism And Susceptibility To Noise-Induced Hearing Loss

Purpose:To explore the association between STAT3 gene polymorphisms(rs4796793,rs1053023,rs1053005,rs1053004,and rs3744483)and the susceptibility of Chinese Han population to noise-induced hearing loss(NIHL),in order to better prevent NIHL and provide scientific basis.Explore whether miR-452-3p can target the rs1053005 site of STAT3,and the relationship between STAT3 factor and active oxygen in the hair cell line HEI-OC1(House Ear Institute-Organ of Corti 1)under oxidative stress.Explore the expression of STAT3 in the cochlea of noise-induced mice,and explore the potential role of STAT3 in the pathogenesis of noise-induced hearing loss from animal experiments Method:The research subjects in this experiment were 2100 employees who were exposed to noise from an energy company,an auto parts manufacturing plant,and a chemical fiber factory in Jiangsu Province.They all participated in the occupational health examinations organized by their companies.Use questionnaire methods to collect information on age,gender,work time with noise,smoking,drinking,etc.,use on-site epidemiological methods to conduct audiometry on noise-exposed workers,and divide them into hearing loss group(case group)and normal hearing group based on the audiology evaluation results.The study population was screened according to uniform inclusion and exclusion criteria,and matched with the frequency of factors such as age,gender,smoking,drinking,and noise exposure level.Finally,a total of 610 cases and 610 cases in the normal hearing group were included.Crowd data analysis uses SPSS 21.0 software for statistical analysis.The demographic data and the distribution of the five gene loci of STAT3 in the case group and the control group,and the comparison between the genotype groups of each STAT3 locus were performed by theχ2 test;after adjusting for factors such as age,gender,smoking,and alcohol use,multiple Factor Logistic regression analysis of the risk of NIHL among workers with different genotypes of noise.The dual luciferase reporter gene experiment was used to verify whether miR-452-3p and STAT3 were targeted to bind at site rs1053005.Overexpress miR-452-3p in HEI-OC1 cells,and observe the expression of STAT3 mRNA and protein in HEI-OC1 cells.Flow cytometry was used to detect the ROS content in HEI-OC1 cells after overexpression of miR-452-3p to explore the relationship between the change in STAT3 expression and the content of reactive oxygen species in the cells.6-week-old C57BL/6J male mice were exposed to 120 dB white noise for 2 hours to construct a noise mouse model.The Elisa experiment was used to detect the content of oxidative stress product malondialdehyde(MDA)in the serum of mice.Western blotting was performed to determine the expression of STAT3 and apoptosis pathway related proteins in noise mice and normal hearing mice.Result:The mRNA expression level of STAT3 in blood samples of the case group was higher than that of the normal hearing group.The correlation analysis between the dominant model and co-dominant model at rs1053023 and rs1053005 loci of STAT3 and NIHL was statistically significant(P<0.05).In the stratified analysis of genotype polymorphism and NIHL risk,for rs1053023 and rs1053005,males carrying the combined TC/CC genotype have a 0.545 and 0.531 times higher risk of NIHL than individuals carrying the TT genotype.In the stratified analysis of smoking and drinking,people who regularly smoke and do not drink TC/CC genotype are more likely to develop NIHL than individuals who carry TT genotype(P<0.05).In the results of the dual luciferase reporter gene experiment,the relative fluorescence intensity value in the cells overexpressing miR-452-3p was significantly reduced,suggesting that miR-452-3p was targeted to bind to the rs1053005 site of STAT3.After overexpression of miR-452-3p,the expression of STAT3 mRNA and protein in HEI-OC1 cells was significantly reduced.After overexpression of miR-452-3p,the level of ROS in HEI-OC1 cells under oxidative stress was significantly reduced.The MDA content in the serum of mice in the hearing loss group was higher than that in the normal hearing group(P<0.05)and the apoptotic pathway of the cochlear tissue was activated.In the cochlea tissue of noisy mice,the expression level of STAT3 protein increased.Conclusion:The susceptibility of the single nucleotides rs1053023 and rs1053005 of STAT3 to NIHL is statistically significant,and the C alleles at the two loci may become the susceptibility genes of NIHL.miR-452-3p targets and regulates the expression of STAT3 and alleviates the generation of ROS in hair cells,providing genetic basis for subsequent studies on the specific biological processes that STAT3 participates in the cochlea.Noise exposure induced an increase in the level of oxidative stress in mice and activation of the cochlear apoptotic pathway.In population data and in vivo animal experiments,the expression of STAT3 in the noise-exposed group increased to a certain extent,combined with the targeted regulation of STAT3 by miR-452-3p,indicating that the miR-452-3p/STAT3 regulatory axis will play an important role in the prevention of NIHL.