Difference Analysis Of Serum Expression Of MiRNAs In Workers Of Occupational Noise-induced Deafness

Objective: To screen the differentially expressed circulating mi RNAs between workers with occupational noise-induced hearing loss(NIHL) and workers with non-NIHL, then to analyze the target genes of differentially expressed mi RNAs. The general purpose of this study is to provide experimental support for the mechanisms underlying the pathogenesis of NIHL, and further, for its prevention and treatment.Methods:1. Collected, numbered and aliquoted serum samples and snapped frozen at-80 ℃for long-term storage. Recorded the information of blood sample providers into database,then extract the information related to qualified serum samples from the database. 2.Extracted and purified mi RNAs from serum samples, and spectrophotometer was use for the quality testing. 3. Preparation of blocking and hybridization buffer, and following steps like sample preparation, hybridization, chiping, chip washing, scanning, chip data extraction,were perform, respectively. Finally, the experimental results from mi RNA hybridization were analyzed to find out the differentially expressed mi RNAs. 4. Verified the results of chip hybridization by fluorescence quantitative PCR. Then collected and conformed the predicted target genes of these mi RNAs.Results:1. Sera from all blood samples were extracted, recorded and stored. A blood sample database was established. Total 6 serum samples(control groups and deaf groups each for 3)were selected for the following studies. 2. After extraction, quality testing of mi RNAs,deafness samples(1-3), and control group samples(1-3) meet the criteria of microarray experiments. 3. Quality test shows that samples are of integrity and good enough for the hybridization reaction. Chips experiments showed that there are 8 differentially of mi RNAs : hsa-mi R-1229-5p, hsa-mi R-3162-5p, hsa-mi R-4459, hsa-mi R- 483-5p,hsa-mi R-6087, hsa-mi R-6088, hsa-mi R-638, hsa-mi R-4484. 4. Fluorescence quantitative PCR showed that the expressions of hsa-mi R-1229-5p and hsa-mi R-483-5p are higher in NIHL groups compared to non-NIHL. Prediction of target genes about hsa-mi R-1229-5p are MAPK1, SLC12A6, SLC25A24, SLC30A8 and hsa-mi R-483-5p is SRF.Conclusion:1. Microarray hybridization found 8 mi RNAs with differential expression: hsami R-1229-5p, hsa-mi R-3162-5p, hsa-mi R-4459, and hsa-mi R-483-5p, and hsa-mi R-6087,and hsa-mi R-6088, and hsa-mi R-638, and hsa-mi R-4484. 2. The enhanced expression of hsa-mi R-1229-5p and hsa-mi R-483-5p are conformed in workers with NIHL in comparison with non-NIHL workers. Prediction of target genes about hsa-mi R-1229-5p are MAPK1,SLC12A6, SLC25A24, SLC30A8 and hsa-mi R-483-5p is SRF.