Jung Jin Woo, Li Hui, Lee Jung Hun, Hwang Yu-Jung, Dan Kisoon, Park Moo Kyun, Han Dohyun, Suh Myung-Whan
Proteomics Core Facility, Transdisciplinary Research and Collaboration, Biomedical Research Institute, Seoul National University Hospital, Seoul, South Korea.
Department of Otorhinolaryngology-Head and Neck Surgery, Seoul National University Hospital, Seoul, South Korea.
Front Pharmacol. 2023 Jan 19;14:1081724. doi: 10.3389/fphar.2023.1081724. eCollection 2023.
Until recently, the most standard treatment for sensorineural or sudden hearing loss, which is caused by inner ear damage or deterioration, has been systemic oral steroid administration. In recent, intratympanic steroid injections such as dexamethasone have been used for the treatment of sudden hearing loss as well. It is injected into the tympanic cavity through its membrane and is expected to diffuse over the round window located between the tympanic cavity and the inner ear. However, in clinical situations, the delivery time of steroids to the inner ear is shorter than 24 h, which does not allow for a sufficient therapeutic effect. Therefore, we applied a previously invented dual viscosity mixture vehicle (DVV) for intratympanic dexamethasone to a guinea pig model, which could reduce the side effects of systemic steroid administration with sufficient dwelling time for the treatment of hearing loss, and we investigated the physiological changes with a global proteomic approach. In this study, we extracted perilymph in three different conditions from guinea pigs treated with dexamethasone-embedded DVV, dexamethasone mixed in saline, and control groups to compare proteomic changes using tandem mass spectrometry analysis. After liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) analysis, we first identified 46 differentially expressed proteins (DEPs) that were statistically significant after one-way ANOVA multiple-sample test. We also performed pairwise comparisons among each group to identify DEPs closely related to the treatment response of dexamethasone-embedded DVV. Gene ontology enrichment analysis showed that these DEPs were mostly related to inflammation, immune, actin remodeling, and antioxidant-related processes. As a result, the proteome changes in the DVV-treated groups revealed that most upregulated proteins activate the cell proliferation process, and downregulated proteins inhibit apoptosis and inflammatory reactions. Moreover, the reactive oxygen process was also regulated by DEPs after DVV treatment.
直到最近，对于由内耳损伤或退化引起的感音神经性或突发性听力损失，最标准的治疗方法一直是全身性口服类固醇给药。最近，诸如地塞米松之类的鼓室内类固醇注射也已用于治疗突发性听力损失。它通过鼓膜注入鼓室，并有望扩散到位于鼓室和内耳之间的圆窗。然而，在临床情况下，类固醇向内耳的递送时间短于24小时，这无法产生足够的治疗效果。因此，我们将先前发明的用于鼓室内地塞米松的双粘度混合载体（DVV）应用于豚鼠模型，该模型可以减少全身性类固醇给药的副作用，并具有足够的停留时间来治疗听力损失，并且我们采用全局蛋白质组学方法研究了生理变化。在本研究中，我们从用嵌入地塞米松的DVV、地塞米松与盐水混合处理的豚鼠以及对照组中提取了三种不同条件下的外淋巴，以使用串联质谱分析比较蛋白质组变化。经过液相色谱-串联质谱（LC-MS/MS）分析后，我们首先鉴定出46种差异表达蛋白（DEP），在单向方差分析多样本检验后具有统计学意义。我们还在每组之间进行了成对比较，以鉴定与嵌入地塞米松的DVV治疗反应密切相关的DEP。基因本体富集分析表明，这些DEP大多与炎症、免疫、肌动蛋白重塑和抗氧化相关过程有关。结果，DVV处理组中的蛋白质组变化表明，大多数上调的蛋白质激活细胞增殖过程，而下调的蛋白质抑制细胞凋亡和炎症反应。此外，DVV处理后，活性氧过程也受到DEP的调节。
