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目的利用抑制消减杂交技术(SSH)筛选分析噪声性听觉损伤易感与不易感豚鼠耳蜗差异表达基因。方法易感组与不易感组豚鼠耳蜗组织利用Trizol法分别提取总RNA;SmartTM技术分别合成cDNA;SSH筛选差异表达基因,并进行测序和生物信息学分析。结果筛选噪声性听觉损伤(NIHL)易感与不易感豚鼠耳蜗的明显差异表达基因片段148条,包括易感组豚鼠耳蜗表达上调73条和下调75条。同源性大于80%且匹配碱基大于100 bp的相关序列共34条,其中上调7条,包括多聚泛素基因、β珠蛋白和α珠蛋白等;下调27条,包括线粒体基因组(mtDNA)、热休克蛋白(HSP)和细胞周期非特异性因子1等。结论SSH是一种高效的筛选差异表达基因的方法,mtDNA和HSP与NIHL及其易感性的发生相关。
OBJECTIVE: To screen differentially expressed genes of cochleas in guinea pigs susceptible to and susceptible to noise-induced hearing impairment using SSH screening. Trizol method was used to extract the total RNA of the cochlear in the susceptible group and the non-susceptible group, respectively. The cDNAs were synthesized by SmartTM technology, and the differentially expressed genes were screened by SSH. Sequencing and bioinformatics analysis were also performed. Results A total of 148 genes were significantly differentially expressed in guinea pig cochleas of susceptible and non-susceptible guinea pigs, including 73 up-regulated and 75 down-regulated in cochleas of susceptible guinea pigs. A total of 34 related sequences with homology greater than 80% and matching base greater than 100 bp were up-regulated by 7 genes including polyubiquitin gene, β-globin and α-globin, down-regulated by 27, including mitochondrial genomes ), Heat shock protein (HSP) and cell cycle non-specific factor 1 and so on. Conclusion SSH is an efficient method for screening differentially expressed genes. The correlation of mtDNA and HSP with NIHL and its susceptibility is significant.