联合生物信息学分析与RNA测序揭示扣带回中与耳鸣相关的关键分子

文献信息

中文摘要

耳鸣源于适应不良的可塑性和异常突触功能，不仅发生在听觉通路，也发生在边缘系统区域。其中，扣带回在塑造耳鸣患者常见的情感负担和感觉过滤障碍方面发挥关键作用。然而，该区域内具体发挥作用的分子仍知之甚少，且现有大多数生物信息学预测尚未在活体系统中得到验证。在此，我们将多数据库挖掘与体内转录组分析相结合，以识别并验证扣带回中与耳鸣相关的关键蛋白和通路。我们构建并比较了代表听觉感知、声学挑战和耳鸣状态的蛋白质-蛋白质相互作用网络，从而锁定候选枢纽分子。为验证这些候选分子，我们使用了噪声诱导的耳鸣小鼠模型，通过听惊反射的前脉冲抑制（GPIAS）确认表型，然后对扣带回组织进行RNA-seq和qPCR。生物信息学筛选指向AKT1和TNF为中心枢纽，提示存在炎症-存活信号轴。在动物实验中，RNA-seq显示耳鸣组Akt1转录水平明显升高，功能富集分析指向细胞外基质组织、细胞黏附和细胞因子受体通路的显著变化。qPCR进一步证实了Akt1上调。综上所述，我们的整合策略确立AKT1和TNF为扣带回耳鸣网络中拓扑中心的枢纽蛋白，并在体内验证了Akt1上调，为未来研究提供了潜在的生物标志物候选和治疗靶点。

英文摘要

Tinnitus arises from maladaptive plasticity and abnormal synaptic function, not only in auditory pathways but also in limbic regions. Among these, the cingulate gyrus plays a pivotal part in shaping the emotional burden and sensory filtering disturbances often seen in tinnitus patients. Yet, the specific molecular players operating within this region remain poorly understood, and most existing bioinformatic predictions have not been tested in living systems. Here, we combined multi-database mining with in vivo transcriptomic profiling to identify and validate key proteins and pathways linked to tinnitus in the cingulate gyrus. We built and compared protein-protein interaction networks representing auditory perception, acoustic challenge, and tinnitus states, thereby pinning down candidate hub molecules. To verify these candidates, we used a noise-induced mouse model of tinnitus, confirmed the phenotype via gap prepulse inhibition of acoustic startle (GPIAS), and then performed RNA-seq and qPCR on cingulate gyrus tissue. The bioinformatic screens pointed to AKT1 and TNF as central hubs, suggestive of an inflammatory-survival signaling axis. In our animal experiments, RNA-seq showed a clear rise in Akt1 transcript levels in the tinnitus group, and functional enrichment analysis pointed to marked changes in extracellular matrix organization, cell adhesion, and cytokine receptor pathways. qPCR further corroborated the Akt1 upregulation. Taken together, our integrative strategy establishes AKT1 and TNF as topologically central hub proteins in the cingulate gyrus tinnitus network, and validates Akt1 upregulation in vivo, offering potential biomarker candidates and therapeutic leads for future studies.