噪声性听力损失大鼠耳蜗miRNA调控网络分析  

Analysis of microRNA regulatory network in rat cochlea with noise induced hearing loss by the next generation sRNA sequencing technology

作　　者：于碧鲲 崔鹂 夏源[2] 王致[3] 王军义[2] 

机构地区：[1]深圳龙岗区疾病控制中心,广东深圳518172 [2]广东药学院公共卫生学院,广东广州510310 [3]广州市职业病防治院,广东广州510620

出　　处：《现代预防医学》2016年第1期124-127,135,共5页Modern Preventive Medicine

基　　金：国家自然科学基金(81202179);广东省自然科学基金(S2012040006490);广东药学院科技处-附属第一医院联合自然科学培育基金(GYFYLH201326)

摘　　要：目的通过噪声性听力损失(NIHL)耳蜗差异性mi RNA构建的基因调控网络,分析NIHL耳蜗的重要基因及功能,探讨NIHL的损伤机制。方法 6只SPF及SD大鼠通过110 d B SPL(6 h/d、24 d)的高斯白噪声暴露建立NIHL模型,6只SPF及SD大鼠非噪声暴露作对照,噪声暴露后7 d取实验大鼠耳蜗组织进行s RNA深度测序,分析差异性表达的mi RNA,以差异最显著的8个mi RNA依据靶基因数据库Target Scan、mi Randa和mi RDB共同预测的靶基因建立差异性表达mi RNA的靶基因集,结合蛋白互作数据库构建mi RNA调控网络,统计网络中每个基因相连基因的个数,确定相连基因个数大于30的基因作为NIHL大鼠耳蜗的重要基因,通过Gene Cards数据库对重要基因进行功能注释。结果 (1)噪声暴露组大鼠永久性听阈位移水平(PTS)明显高于对照组(秩和检验:Z值-2097,P<0.01);(2)s RNA测序结果显示2组耳蜗差异性mi RNA有148个[|log2(Fold Change)|>1],差异性最显著的8个mi RNA为:rno-mi R-378b、rno-mi R-211-5p、rno-mi R-133b-3p、rno-mi R-29c-3p、rno-let-7c-2-3p、rno-mi R-674-5p、rno-mi R-495、rno-mi R-3068-3p;(3)mi RNA调控网络中的重要基因有11个:VEGFA、TNF、EGFR、FOS、NR3C1、AGT、CREBBP、PTK2、CD44、STAT3、IGF1。结论 NIHL大鼠耳蜗的11个重要基因通过调控细胞增殖与分化、控制细胞凋亡、改善组织细胞营养代谢、介导组织炎性反应以改善和修复耳蜗组织细胞。Objective Gene regulatory networks which based on different miRNA in cochlear with noise induced hearing loss （NIHL） was used to analyze the damage mechanisms of NIHL and the important gene and function of cochlear with NIHL. Methods 6 SPF SD rats were exposed to Gaussian white noise at ll0dB SPL 6 h a day for 28 d. Other 6 SPF SD rats were used as control group. We used the deep-sequencing technique to analyze the difference expression of miRNA in cochlear 7 days after exposure. Eight most significantly different miRNA and predicted target gene based on target genes database TargetScan, miRanda and miRDB to establish a difference expression of micromas gene cluster. We constructed miRNA regulation network by using protein interaction database. According to the statistics of each connected genes number in network, we find the genes which is greater than 30. These genes were presented functional annotation as important genes in rat cochlea with NIHL by GeneCards database. Results ①There had significant difference during groups, increased level of PTS were observed in exposed group with higher intensity noise （rank-sum test： Z value -2097, P〈0.01）. ②sRNA sequence indicate there are 148 different genes in the two groups [llog2 （Fold Change）l〉1]. Eight most significant difference miRNA is： ruo-miR-378b, mo-miR-211-5p, mo-miR-133b-3p, mo-miR-29c-3p, rno-let-7c-2-3p, rno-miR-674-5p, mo-miR-495, ruo-miR-3068-3p. ③ Eleven important genes in miRNA regulatory network is： VEGFA, TNF, EGFR, FOS, NR3C1, AGT, CREBBP, PTK2, CD44, STAT3, IGF1. Conclusion The 11 important genes in rat cochlea with NIHL through regulating cell proliferation and differentiation, controlling cell apoptosis, improving nutrition metabolism and mediating inflammatory reaction to improve and repair the cochlear cells.

关 键 词：噪声性听力损失 下一代sRNA测序 MicroRNA调控网络 耳蜗 

分 类 号：R181.24[医药卫生—流行病学]