Department of Respiratory Medicine, Huai'an Second People's Hospital, The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, China.
Gastroenterology Department, Nanjing Pukou District Central Hospital, Pukou Branch of Jiangsu People's Hospital, Nanjing, China.
J Appl Toxicol. 2022 Apr;42(4):671-682. doi: 10.1002/jat.4249. Epub 2021 Oct 15.
Asthma progression is involved in airway epithelial dysfunction, airway inflammatory response, and mucus hypersecretion. Euxanthone has been found to exhibit cytotoxic activity on several human diseases, such as neurological disorders and cancers. Our study aimed to explore the influence of euxanthone on lipopolysaccharide (LPS)-induced injury, inflammatory response, and mucin 5AC (MUC5AC) hypersecretion in human airway epithelial cells (AECs). Network pharmacology analysis was carried out to analyze the drug targets and key pathways of euxanthone against asthma. Cell injury was evaluated by CCK-8, Lactate dehydrogenase (LDH) release assay, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The production of interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1), and MUC5AC was measured using enzyme-linked immunosorbent assay (ELISA). MUC5AC mRNA expression was detected by qRT-PCR. Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) protein expression was examined by western blot analysis. Venn diagram showed 14 overlapping targets between euxanthone and asthma. According to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, we focused on TLR signaling pathway. LPS exposure evoked viability reduction, increased LDH release and apoptosis, and induced production of inflammatory cytokines (IL-6, IL-8, and MCP-1) and MUC5AC hypersecretion in human AECs, which were alleviated by euxanthone. Mechanistically, we validated that euxanthone attenuated LPS-induced activation of TLR4/MyD88 pathway in AECs. Moreover, inhibition of the TLR4/MyD88 pathway enhanced the inhibitory effect of euxanthone on LPS-induced cell injury, inflammatory response and MUC5AC expression. In conclusion, euxanthone attenuated LPS-induced cell injury, inflammatory response, and MUC5AC expression in AECs by inhibiting the activation of TLR4/MyD88 pathway.
哮喘的进展涉及气道上皮功能障碍、气道炎症反应和黏液高分泌。柚皮素已被发现对几种人类疾病具有细胞毒性作用，如神经紊乱和癌症。我们的研究旨在探讨柚皮素对人呼吸道上皮细胞（AEC）中脂多糖（LPS）诱导的损伤、炎症反应和粘蛋白 5AC（MUC5AC）过度分泌的影响。通过网络药理学分析研究柚皮素治疗哮喘的药物靶点和关键途径。通过 CCK-8、乳酸脱氢酶（LDH）释放试验和末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）试验评估细胞损伤。采用酶联免疫吸附试验（ELISA）测定白细胞介素（IL）-6、IL-8、单核细胞趋化蛋白-1（MCP-1）和 MUC5AC 的产生。通过 qRT-PCR 检测 MUC5AC mRNA 表达。通过 Western blot 分析检测 Toll 样受体 4（TLR4）和髓样分化因子 88（MyD88）蛋白表达。Venn 图显示柚皮素和哮喘之间有 14 个重叠靶点。根据京都基因与基因组百科全书（KEGG）通路富集分析，我们重点关注 TLR 信号通路。LPS 暴露可引起人 AEC 活力降低、LDH 释放和凋亡增加，并诱导炎症细胞因子（IL-6、IL-8 和 MCP-1）和 MUC5AC 的过度分泌，柚皮素可减轻这些变化。机制上，我们验证了柚皮素可减弱 AEC 中 LPS 诱导的 TLR4/MyD88 通路的激活。此外，抑制 TLR4/MyD88 通路可增强柚皮素对 LPS 诱导的细胞损伤、炎症反应和 MUC5AC 表达的抑制作用。综上所述，柚皮素通过抑制 TLR4/MyD88 通路的激活，减轻 LPS 诱导的 AEC 细胞损伤、炎症反应和 MUC5AC 表达。
