【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。更多还原

【Abstract】 Objective Gene regulatory networks which based on different mi RNA 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. Methods6 SPF SD rats were exposed to Gaussian white noise at 110 d B 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 mi RNA in cochlear 7 days after exposure.Eight most significantly different mi RNA and predicted target gene based on target genes database Target Scan, mi Randa and mi RDB to establish a difference expression of micrornas gene cluster. We constructed mi RNA 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 than30. These genes were presented functional annotation as important genes in rat cochlea with NIHL by Gene Cards database. Results1 There 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). 2s RNA sequence indicate there are 148 different genes in the two groups[|log2(Fold Change)|>1]. Eight most significant difference mi RNA is: rno-mi R-378 b, 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 Eleven important genes in mi RNA 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.更多还原