Background: Acute otitis media (AOM) is highly prevalent among preschool children and can lead to chronic otitis media or severe complications if untreated. The pathogenesis of AOM is closely tied to the host's immune response, yet the expression profile of cytokines, which mediate these responses, remains poorly defined.
背景： 急性中耳炎（AOM）在学龄前儿童中非常普遍，若未治疗可能导致慢性中耳炎或严重并发症。急性中耳炎的发病机制与宿主的免疫反应密切相关，但介导这些反应的细胞因子表达谱仍不明确。

Purpose: This study aims to identify cytokines involved in AOM immune responses and to elucidate their roles at different stages of the disease.
目的： 本研究旨在识别参与 AOM 免疫反应的细胞因子，并阐明它们在疾病不同阶段的作用。

Methods: We induced AOM in mice by injecting nontypeable Haemophilus influenzae (NTHi) through the tympanic membrane. Middle ear tissues were collected for analysis. PCR Array and Protein Array identified cytokines at 6 and 12 hours (h) post-infection. qPCR and Luminex multiplex detection technology were then used to track cytokine expression at various key time points, allowing us to speculate on their roles in the immune process.
方法： 我们通过通过鼓膜注射非分型流感嗜血杆菌（NTHi）诱导小鼠中耳中耳炎。采集了中耳组织进行分析。PCR 阵列和蛋白质阵列在感染后 6 小时和 12 小时时鉴定了细胞因子。随后使用 qPCR 和 Luminex 多重检测技术追踪细胞因子在不同关键时刻的表达，使我们能够推测它们在免疫过程中的作用。

Results: PCR Array identified 13 cytokines with significant mRNA differences at 6 h. Protein Array identified 9 differentially expressed cytokines at 12 h qPCR tracked mRNA expression at key time points (6 h, 12 h, 1day (d), 2 d, 3 d and 7 d), revealing a total of 20 cytokines. Luminex detected 31 cytokines at key time points (12 h 1 d, 2 d and 5 d), 10 of which were previously mentioned. Cytokines were classified into three distinct categories according to their expression trends: early-phase (n = 13), progressive-phase (n = 20), and terminal-phase (n = 4) cytokines.
结果： PCR 阵列鉴定出 13 种在 6 小时时 mRNA 显著差异的细胞因子。蛋白阵列在 12 小时识别出 9 种差异表达的细胞因子，qPCR 追踪关键时间点（6 小时、12 小时、1 天（d）、2 天、3 天和 7 天的 mRNA 表达，共发现 20 种细胞因子。Luminex 在关键时间点（12 小时、1 天、2 天和 5 天）检测到 31 种细胞因子，其中 10 种已提及。细胞因子根据表达趋势被分为三类：早期阶段（n=13）、渐进阶段（n=20）和末期（n=4）。

Conclusion: Cytokines play distinct and crucial roles at each stage of AOM, with potential interactions among those highly expressed in the same stage. 
结论： 细胞因子在 AOM 的每个阶段都扮演着不同且关键的角色，且同一阶段中表达较多的细胞因子之间可能存在相互作用。