前胡素A抑制poly(I:C)诱导的RAW264.7细胞中NF-κB通路的激活和炎症因子的表达。

Jiayan Hu, Roujun Liu, Zhouxin Yang, Xinyu Pan, Yuanjing Li, Yanghui Gong, Dongyang Guo
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摘要

前胡素A(PA)是一种天然香豆素化合物,具有显著的抗炎作用。在本研究中,我们评估了PA对聚肌苷酸-聚胞苷酸(poly(I:C))诱导的RAW 264.7小鼠巨噬细胞的抗炎作用。将poly(I:C)诱导的RAW 264.7小鼠巨噬细胞用或不用PA处理,测定其活力以筛选PA的工作溶液。应用RNA测序分析差异表达基因(DEGs)。进行了基因本体论(GO)和京都基因与基因组百科全书(KEGG)分析。采用酶联免疫吸附试验(ELISA)、定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹法检测白细胞介素(IL)-1β、血红素加氧酶1(HMOX1)、前列腺素内过氧化物合成酶2(PTGS2)、ATP结合盒亚家族A成员1(Abca1)和NF-κB相关蛋白的表达。结果,PA在1、2、3、4和5 μM轻微影响细胞活力,而PA在6和7 μM显著抑制细胞活力。GO和KEGG分析结果显示,DEG主要富集在与炎症信号相关的通路中。通过进一步分析,我们获得了PA的五个可能靶点,并验证了PA在poly(I:C)诱导的RAW264.7细胞中抑制IL-1β、HMOX1、PTGS2和Abca1的表达以及NF-κB通路的激活。总之,PA可能抑制poly(I:C)诱导的RAW264.7细胞中炎症相关基因的表达,这证明了其作为对抗病毒相关疾病的药物的潜力。
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Praeruptorin A inhibits the activation of NF-κB pathway and the expressions of inflammatory factors in poly (I:C)-induced RAW264.7 cells

Praeruptorin A (PA), a natural coumarin compound, has significant anti-inflammatory effects. In this study, we evaluate the anti-inflammatory effect of PA on RAW 264.7 mouse macrophages induced by Polyinosinic acid-polycytidylic acid (poly (I:C)). RAW 264.7 mouse macrophages induced by poly (I:C) were treated with or without PA, the viability of which was determined to screen working solution of PA. RNA-sequencing was applied to analyze the differentially expressed genes (DEGs). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were carried out. The expressions of interleukin (IL)-1β, heme oxygenase 1 (HMOX1), prostaglandin-endoperoxide synthase 2 (PTGS2), ATP binding cassette subfamily A member 1 (Abca1) and NF-κB-related proteins were measured by enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. As a result, PA at 1, 2, 3, 4 and 5 μM slightly affected cell viability, while PA at 6 and 7 μM significantly inhibited cell viability. GO and KEGG analysis results revealed that DEGs were mainly enriched in the pathways related to inflammatory signaling. Through further analysis, we obtained five possible targets of PA, and verified that PA inhibited the expressions of IL-1β, HMOX1, PTGS2 and Abca1 as well as the activation of NF-κB pathway in poly (I:C)-induced RAW264.7 cells. To summarize, PA may inhibit expressions of the inflammation-related genes in poly (I:C)-induced RAW264.7 cells, which demonstrates its potential as a drug against virus related diseases.

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