脂氧素受体激动剂 BML-111 对香烟烟雾提取物诱导的 RAW264.7 细胞中巨噬细胞极化和炎症的影响。
Effect of the Lipoxin Receptor Agonist BML-111 on Cigarette Smoke Extract-Induced Macrophage Polarization and Inflammation in RAW264.7 Cells.
机构信息
Department of Pathology, Basic Medical College of Nanchang University, Nanchang, Jiangxi, People's Republic of China.
Department of Critical Care Medicine/ICU (Intensive Care Unit), Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China.
出版信息
Int J Chron Obstruct Pulmon Dis. 2023 May 19;18:919-932. doi: 10.2147/COPD.S395569. eCollection 2023.
BACKGROUND
Macrophages are known to play a crucial role in the chronic inflammation associated with Chronic Obstructive Pulmonary Disease (COPD). BML-111, acting as a lipoxin A4 (LXA4) receptor agonist, has shown to be effective in protecting against COPD. However, the precise mechanism by which BML-111 exerts its protective effect remains unclear.
METHODS
In order to establish a cell model of inflammation, cigarette smoke extract (CSE) was used on the RAW264.7 cell line. Afterwards, an Enzyme-linked immunosorbent assay (ELISA) kit was employed to measure concentrations of tumor necrosis factor-α (TNF-α), interleukin-1beta (IL-1β), interleukin-18 (IL-18), and interleukin-10 (IL-10) in the cell supernatants of the RAW264.7 cells.In this study, we examined the markers of macrophage polarization using two methods: quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. Additionally, we detected the expression of Notch-1 and Hes-1 through Western blotting.
RESULTS
BML-111 effectively suppressed the expression of pro-inflammatory cytokines TNF-α, IL-1β, and IL-18, as well as inflammasome factors NLRP3 and Caspase-1, while simultaneously up-regulating the expression of the anti-inflammatory cytokine IL-10 induced by CSE. Moreover, BML-111 reduced the expression of iNOS, which is associated with M1 macrophage polarization, and increased the expression of Arg-1, which is associated with M2 phenotype. Additionally, BML-111 downregulated the expression of Hes-1 and the ratio of activated Notch-1 to Notch-1 induced by CSE. The effect of BML-111 on inflammation and macrophage polarization was reversed upon administration of the Notch-1 signaling pathway agonist Jagged1.
CONCLUSION
BML-111 has the potential to suppress inflammation and modulate M1/M2 macrophage polarization in RAW264.7 cells. The underlying mechanism may involve the Notch-1 signaling pathway.
背景
众所周知，巨噬细胞在慢性阻塞性肺疾病（COPD）相关的慢性炎症中发挥着关键作用。BML-111 作为脂氧素 A4（LXA4）受体激动剂，已被证明在预防 COPD 方面有效。然而，BML-111 发挥其保护作用的确切机制尚不清楚。
方法
为了建立炎症细胞模型，我们使用香烟烟雾提取物（CSE）处理 RAW264.7 细胞系。随后，采用酶联免疫吸附试验（ELISA）试剂盒测量 RAW264.7 细胞上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-18（IL-18）和白细胞介素-10（IL-10）的浓度。在这项研究中，我们使用两种方法检查了巨噬细胞极化的标志物：实时定量聚合酶链反应（qRT-PCR）和 Western blot 分析。此外，我们通过 Western blot 检测了 Notch-1 和 Hes-1 的表达。
结果
BML-111 可有效抑制 CSE 诱导的促炎细胞因子 TNF-α、IL-1β 和 IL-18 以及炎症小体因子 NLRP3 和 Caspase-1 的表达，同时上调 CSE 诱导的抗炎细胞因子 IL-10 的表达。此外，BML-111 降低了与 M1 巨噬细胞极化相关的 iNOS 的表达，增加了与 M2 表型相关的 Arg-1 的表达。此外，BML-111 下调了 CSE 诱导的 Hes-1 表达和活化 Notch-1 与 Notch-1 的比值。Jagged1 给药逆转了 BML-111 对炎症和巨噬细胞极化的作用。
结论
BML-111 具有抑制 RAW264.7 细胞炎症和调节 M1/M2 巨噬细胞极化的潜力。其潜在机制可能涉及 Notch-1 信号通路。
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