欧前胡素通过降低 NF-κB p65 磷酸化抑制 LPS 诱导的骨髓来源巨噬细胞活化。
Imperatorin inhibits LPS-induced bone marrow-derived macrophages activation by decreased NF-κB p65 phosphorylation.
机构信息
Department of Pulmonology, Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, Zhejiang, China.
Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University, School of Medicine, Hangzhou, China.
出版信息
Immunopharmacol Immunotoxicol. 2023 Oct;45(5):581-588. doi: 10.1080/08923973.2023.2196603. Epub 2023 Apr 11.
BACKGROUND
Imperatorin (IMP) is a secondary metabolite of plants and is the most abundant in Angelica dahurica. Previous studies showed that IMP exhibited anti-inflammatory activity in RAW264.7 cell line. Here, we aim to investigate the roles and mechanisms of IMP in bone marrow-derived macrophages (BMDMs), in view of the difference between primary macrophages and cell lines.
METHODS
BMDMs were stimulated with LPS for the inflammation model. Flow cytometry was performed with BMDMs treated with different doses of IMP (0-20mg/L) within staining Annexin V-APC for 5 min. The cytokines and inflammatory mediators were detected by RT-PCR or ELISA. RNA-seq was performed in IMP-treated BMDMs or control, stimulated with LPS for 6h. Western blotting is carried out to determine the phosphorylation of p65, ERK1/2, JNK1, p38, and Akt.
RESULTS
Our results showed that IMP inhibited IL-12p40, IL-6, TNF-α and IL-1β in LPS-stimulated BMDMs. RNA-seq analysis suggested that IMP inhibits Toll-like receptor signaling pathway (KEGG), TNF signaling pathway (KEGG), NF-κB signaling pathway (KEGG), Inflammatory Response (GO). In addition, IMP inhibited , , , (COX-2) expression in mRNA level. Finally, we found decreased phosphorylation of NF-κB p65 in IMP-treated BMDMs, after stimulated with LPS.
CONCLUSION
IMP inhibits IL-12p40, IL-6, TNF-α, and IL-1β expression in LPS-stimulated BMDMs. IMP inhibits macrophage activation, which maybe resulted in decreased phosphorylation of NF-κB p65. Furthermore, IMP may protect against the progress of inflammatory-related diseases.
背景
欧前胡素（IMP）是植物的次生代谢产物，在白芷中含量最丰富。先前的研究表明，IMP 在 RAW264.7 细胞系中表现出抗炎活性。在这里，我们旨在研究 IMP 在骨髓来源的巨噬细胞（BMDMs）中的作用和机制，鉴于原代巨噬细胞和细胞系之间的差异。
方法
用 LPS 刺激 BMDMs 建立炎症模型。用不同剂量的 IMP（0-20mg/L）处理 BMDMs 5 分钟，通过流式细胞术对其进行染色 Annexin V-APC。通过 RT-PCR 或 ELISA 检测细胞因子和炎症介质。用 LPS 刺激 IMP 处理或对照的 BMDMs 6h 后进行 RNA-seq 分析。通过 Western blot 确定 p65、ERK1/2、JNK1、p38 和 Akt 的磷酸化。
结果
结果表明，IMP 抑制 LPS 刺激的 BMDMs 中 IL-12p40、IL-6、TNF-α 和 IL-1β 的产生。RNA-seq 分析表明，IMP 抑制 Toll 样受体信号通路（KEGG）、TNF 信号通路（KEGG）、NF-κB 信号通路（KEGG）、炎症反应（GO）。此外，IMP 抑制 LPS 刺激的 BMDMs 中 COX-2 基因的表达。最后，我们发现 LPS 刺激后，IMP 处理的 BMDMs 中 NF-κB p65 的磷酸化减少。
结论
IMP 抑制 LPS 刺激的 BMDMs 中 IL-12p40、IL-6、TNF-α 和 IL-1β 的表达。IMP 抑制巨噬细胞激活，这可能导致 NF-κB p65 的磷酸化减少。此外，IMP 可能有助于预防炎症相关疾病的进展。
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