人脐带间充质干细胞来源的外泌体通过miR-149-3p/NOX2轴减轻C57BL/6小鼠的中耳炎

文献信息

中文摘要

背景： 中耳炎（OM）是一种高度流行的疾病，可导致儿童听力损失。目前OM的临床治疗主要依赖抗生素；然而，抗生素的过度使用加剧了细菌耐药性，构成了全球公共卫生挑战。因此，迫切需要新的OM治疗策略。人脐带间充质干细胞来源的外泌体（hUCMSCs-Exos）已在多种疾病中显示出显著的治疗潜力，但其在OM中的疗效仍不清楚。方法：在本研究中，我们用脂多糖（LPS）诱导C57BL/6小鼠建立OM小鼠模型。OM小鼠通过两种给药方式——鼓膜（TM）和尾静脉（TV）注射——接受人脐带间充质干细胞来源的外泌体（hUCMSCs-Exos）治疗。结果：我们的结果显示，hUCMSCs-Exos显著减轻了小鼠的OM，表现为听觉功能改善、中耳黏膜增厚减少、脓性渗出减少、炎症细胞因子表达下调以及氧化应激减轻。多组学分析显示，HIF-1信号通路中NADPH氧化酶2（NOX2）表达及其上游调节因子miR-149-3p发生显著改变。qRT-PCR和Western blot证实，hUCMSCs-Exos上调miR-149-3p并抑制NOX2表达，从而减少活性氧（ROS）生成，进而减轻中耳炎炎症。此外，体内功能获得和功能缺失实验表明，miR-149-3p是hUCMSCs-Exos调控NOX2表达及下游炎症反应的关键介质。给药途径的比较分析显示，局部TM注射比全身TV递送产生更优的治疗效果。结论：总之，我们的研究结果表明，hUCMSCs-Exos可能通过递送miR-149-3p靶向抑制NOX2表达及其下游炎症级联反应，从而减轻LPS诱导的小鼠OM，为hUCMSCs-Exos作为OM的新型非抗生素治疗策略提供了理论基础。

英文摘要

Otitis media (OM) is a highly prevalent disease leading to hearing loss in children. Current clinical treatment for OM primarily relies on antibiotics; however, the overuse of antibiotics exacerbates bacterial resistance, posing a global public health challenge. Therefore, novel therapeutic strategies for OM are urgently needed. Human umbilical cord mesenchymal stem cell-derived exosomes (hUCMSCs-Exos) have demonstrated significant therapeutic potential in various diseases, but their efficacy in OM remains unclear. In this study, we induced C57BL/6 mice with lipopolysaccharide (LPS) to establish an OM mouse model. OM mice were treated with exosomes derived from human umbilical cord mesenchymal stem cells (hUCMSCs-Exos), through two administration methods, tympanic membrane (TM) and tail vein (TV) injection. Our results showed that hUCMSCs-Exos significantly alleviated OM in mice, as evidenced by improved auditory function, reduced middle ear mucosal thickening, decreased purulent exudation, downregulated expression of inflammatory cytokines, and alleviated oxidative stress. Multi-omics analysis revealed significant alterations in NADPH Oxidase 2 (NOX2) expression and its upstream regulator miR-149-3p within the HIF-1 signaling pathway. qRT-PCR and Western blot confirmed that hUCMSCs-Exos upregulated miR-149-3p and suppressed NOX2 expression, thereby reducing reactive oxygen species (ROS) generation, and consequently alleviating otitis media inflammation. Furthermore, in vivo gain- and loss-of-function experiments demonstrated that miR-149-3p is a key mediator in hUCMSCs-Exos-regulated NOX2 expression and downstream inflammatory responses. Comparative analysis of administration routes revealed that local TM injection yielded superior therapeutic outcomes compared to systemic TV delivery. In summary, our findings indicate that hUCMSCs-Exos may alleviate LPS-induced OM in mice by delivering miR-149-3p to target and inhibit NOX2 expression and its downstream inflammatory cascade, providing a theoretical basis for hUCMSCs-Exos as a novel non-antibiotic therapeutic strategy for OM.