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    基因芯片测序技术分析CIH对子代大鼠内耳基因的调控机制

    • 摘要: 目的 筛选慢性宫内缺氧 ( CIH) 仔鼠内耳的差异表达基因(DEGs)并进行生物信息学的分析。方法 24只SD孕大鼠随机分为正常组和缺氧组,分娩后仔鼠经听力测试,对正常听力仔鼠组(COH组)和耳聋仔鼠(MH组)内耳进性基因芯片测序,并验证结果的可靠性。结果 ①实验总RNA提取的的纯度是合格的②CIH后差异表达基因(Fold Change》2,P value≤0.05) 180 个,上调表达基因88个、下调 92 个。GO 功能注释表明,在分子功能GO分类中:差异基因有电压门控钾通道活性、甲基化CpG结合、主动跨膜转运活性等;在生物学过程:差异基因主要参与蛋白质转运、氧运输、小分子代谢等;细胞组分GO分类:差异基因富集于细胞表面、受体复合物、细胞外间隙、膜固有成分;pathway 富集分析显示:差异表达基因主要在κB信号通路、JNK信号通路、NOD样受体信号通路、Toll样受体信号转导通路等三十多条转录失调的转导通路。③Read-time PCR 验证 DEGs 上调或下调趋 势和 基因芯片检测结果基本一致。结论 妊娠期CIH可上调MH组促细胞凋亡基因(caspase 2、Bak1、Tsc22d3、Ltbr、Casp12、Dapk1等);下调维持离子平衡基因(Slc2a4、GJB2、、CLDN14、、GJB6等)、水通道蛋白(Aqp5、Aqp7)、转录因子(POU4F3)等;下调抑制细胞凋亡基因(Birc3、Nol3、Birc5、Hells、PAK7等·),使κB信号通路、JNK信号通路等转导通路转录失调。

       

      Abstract: objective The differentially expressed genes (DEGs) in the inner ear of rats with chronic intrauterine hypoxia (CIH) were screened and analyzed by bioinformatics. Methods Twenty-four pregnant SD rats were randomly divided into normal group and anoxic group. After delivery, hearing test was performed on the normal hearing group (COH group) and deaf-induced rats (MH group). Result The purity of total experimental RNA extraction is qualified②There were 180 differentially expressed genes (old Change > 2 P value ≤ 0.05), 88 up-regulated genes and 92 down-regulated genes after CIH. GO functional annotation shows that in the molecular functional GO classification, the differential genes have voltage gated potassium channel activity, methylation CpG binding, active transmembrane transport activity, etc. in biological process, differential genes mainly participate in protein transport, oxygen transport and small molecule metabolism, and cell components are classified into GO classification: differentially genes are enriched in cell surface, receptors Complex, extracellular space, and intrinsic components of the membrane; pathway enrichment analysis showed that the differentially expressed genes were mainly more than 30 pathways of transcription disorders such as kappa B signaling pathway, JNK signaling pathway, NOD like receptor signaling pathway and Toll like receptor signal transduction pathway. ③Read-time PCR confirmed that the up-regulation or down-regulation of DEGs was basically consistent with the results of gene chip detection. Conclusion during pregnancy, CIH can up-regulate the apoptosis-promoting gene of MH group, such as Tsc22d3, Tsc22d3, Casp12, Dapk1, etc., down-regulate the ion balance gene Slc2a4, CLDN14, GJB6, aquaporin Aqp7, transcription factor POU4F3, etc. The down-regulation of the apoptosis suppressor gene, Birc3Nol3, Birc5, HellsPak 7 and so on, makes the κ B signaling pathway, JNK signal pathway and other transduction pathways transcriptional imbalance

       

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