主观性耳鸣患者外周血非编码RNA高通量测序初析

Initial analysis of high-throughput sequencing of non-coding RNA in the peripheral blood of patients with subjective tinnitus

  • 摘要: 目的:探寻主观性耳鸣患者的差异表达非编码 RNA(ncRNA)及相应调控的信使 RNA(mRNA)。方法:对 37例主观性耳鸣患者(耳鸣组)和 20 例健康志愿者(健康对照组)外周血样本进行高通量测序分析,以识别差异表达的长链非编码 RNA(lncRNA)、环状RNA(circRNA)和mRNA,并通过公共数据库匹配ncRNA-miRNA-mRNA的调控路径。通过实时荧光定量聚合酶链式反应(qPCR)对另外10例耳鸣组和10例健康对照组外周血样本进行检测,以验证筛选出的ncRNA和mRNA的差异表达是否具有普遍性。结果:在耳鸣组外周血中发现了 13个 lncRNA、596个 circRNA和 38个 mRNA存在显著差异表达。13个差异表达的lncRNA均未能匹配到相应的miRNA,未行后续的mRNA匹配分析。596个差异表达的circRNA成功匹配到58个miRNA和595个mRNA,其中有1个mRNA(TOMM7)显示出差异表达,其对应的circRNA和miRNA分别为circ_0051120和miR-615。qPCR 结果显示,circ_0051120 的表达量在两组间存在显著差异(P<0.05),而 TOMM7 表达无统计学差异(P>0.05)。结论:circ_0051120可能通过调节TOMM7的表达水平,进而影响线粒体自噬过程,并过度激活NF-κB信号通路以及影响神经网络稳定性,从而在主观性耳鸣的发生和维持过程中发挥重要作用。

     

    Abstract: Objective: To investigate the differentially expressed non-coding RNAs (ncRNAs) and their corresponding regulated messenger RNAs (mRNAs) in patients with subjective tinnitus. Methods: High-throughput sequencing analysis was performed on peripheral blood samples from 37 patients with subjective tinnitus (tinnitus group) and 20 healthy volunteers (healthy control group) to identify differentially expressed long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and mRNAs. The regulatory pathways of ncRNA-miRNA-mRNA were matched using public databases. Additionally, quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression of selected ncRNAs and mRNAs in peripheral blood samples from another 10 patients of tinnitus group and 10 healthy controls to verify the universality of the differential expression. Results: Significant differential expression of 13 lncRNAs, 596 circRNAs, and 38 mRNAs were found in the peripheral blood in the tinnitus group. The 13 differentially expressed lncRNAs did not match any corresponding miRNAs, and thus no subsequent mRNA matching analysis was conducted. The 596 differentially expressed circRNAs were successfully matched to 58 miRNAs and 595 mRNAs, among which only one mRNA (TOMM7) exhibited differential expression in this study, with corresponding circRNA and miRNA being circ_0051120 and miR-615, respectively. The qPCR results showed a significant difference in the expression level of circ_0051120 between the two groups (P<0.05), while but there was no significant difference in the expression of TOMM7 (P>0.05). Conclusion: The circ_0051120 may play a crucial role in the occurrence and maintenance of subjective tinnitus by regulating the expression level of TOMM7, thereby affecting mitochondrial autophagy, over-activating the NF- κB signalling pathway, and influencing neural network stability.

     

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