Pneumonia is a pulmonary disease among children. Evodiamine, a traditional Chinese medicine, is known for anti-inflammatory effect. This study aimed to investigate the impact of evodiamine on severe pneumonia-like cells and the underlying mechanism involved. H5N1 and pneumoniae D39 was used to induce severe pneumonia-like conditions in BEAS-2B cells. The cell viability in BEAS-2B cells after treatments with 0, 20, 40, 60, 80, and 100 μM evodiamine was examined using MTT assays. The protein concentrations of inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1β, and Toll-like receptors (TLRs) were measured by enzyme-linked immunosorbent assay methods and the protein and mRNA changes in C/EBPβ/CREB were measured using Real Time-quantitative polymerase chain reaction and Western blot methods. Our results revealed that Evodiamine significantly decreased TNF-α, IL-6, and IL-1β in BEAS-2B cells. Moreover, evodiamine markedly reduced TLR2,3,4 protein expression and the phosphorylated protein of C/EBPβ and CREB. Besides, evodiamine combined with clindamycin exerted more significant effects than clindamycin alone. Taken together, our results demonstrated that evodiamine enhanced the anti-inflammation effect of clindamycin in the BEAS-2B cells infected with H5N1 and pneumoniae D39 through CREB-C/EBPβ signaling pathway.

中文翻译：

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吴茱萸碱通过CREB-C/EBPβ信号通路增强克林霉素在H5N1和肺炎D39感染的BEAS-2B细胞中的抗炎作用

肺炎是一种儿童肺部疾病。吴茱萸碱是一种传统的中药，以抗炎作用而闻名。本研究旨在探讨吴茱萸碱对重症肺炎样细胞的影响及其潜在机制。H5N1 和肺炎 D39 用于在 BEAS-2B 细胞中诱导严重的肺炎样疾病。使用 MTT 测定法检查用 0、20、40、60、80 和 100 μM 吴茱萸碱处理后 BEAS-2B 细胞中的细胞活力。采用酶联免疫吸附法测定炎症细胞因子肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6和IL-1β以及Toll样受体（TLRs）的蛋白浓度，并检测其蛋白和mRNA的变化。使用实时定量聚合酶链反应和蛋白质印迹法测量 C/EBPβ/CREB。我们的结果显示吴茱萸碱显着降低 BEAS-2B 细胞中的 TNF-α、IL-6 和 IL-1β。此外，吴茱萸碱显着降低了 TLR2、3、4 蛋白的表达以及 C/EBPβ 和 CREB ​​的磷酸化蛋白。此外，吴茱萸碱联合克林霉素比单独使用克林霉素效果更显着。总之，我们的结果表明吴茱萸碱通过CREB-C/EBPβ信号通路增强了克林霉素在感染H5N1和肺炎D39的BEAS-2B细胞中的抗炎作用。