Qingfei oral liquid alleviates RSV-induced lung inflammation by promoting fatty-acid-dependent M1/M2 macrophage polarization via the Akt signaling pathway.
Qingfei oral liquid alleviates RSV-induced lung inflammation by promoting fatty-acid-dependent M1/M2 macrophage polarization via the Akt signaling pathway.
复制标题
清肺口服液通过 Akt 信号通路促进脂肪酸依赖性 M1/M2 巨噬细胞极化，减轻 RSV 诱导的肺部炎症。
DOI：
10.1016/j.jep.2022.115637
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发表时间：
2022-11-15
影响因子：
5.4
通讯作者：
Lin, Lili
中科院分区：
文献类型：
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作者：
An, Li;Lu, Mengjiang;Lin, Lili
民族药理学相关性：呼吸道合胞病毒（RSV）是导致婴儿和老年人下呼吸道疾病的常见病原体，并且目前没有可用的疫苗。清肺口服液（QF）是一种传统的中药制剂，临床上已被证明具有抗炎作用。目的：本研究通过脂肪酸依赖性巨噬细胞极化来研究QF是否能抑制RSV诱导的小鼠肺部炎症。材料与方法：BALB/c小鼠感染RSV后，连续4 d灌胃给予低、中、高剂量QF。使用H&E染色和细胞因子测定来评估肺部炎症状态。采用超高效液相色谱/串联质谱法（UPLC-MS/MS）分析QF的活性成分和脂肪酸代谢。通过网络药理学和分子对接研究发现了一条与脂质代谢相关的途径。采用Western印迹法测定ATP-柠檬酸裂解酶（ACLY）、过氧化物酶体增殖激活受体α（PPAR）、Akt蛋白激酶B及其磷酸化形式在Akt信号传导中的水平。流式细胞术定量测定巨噬细胞亚型（M1/M2）的数量，免疫组化检测诱导型一氧化氮合酶（iNOS）和精氨酸酶-1（Arg-1）的表达。结果：在RSV感染小鼠的肺组织中，QF抑制促炎蛋白如白细胞介素-1 β的转录（IL-1 β）、肿瘤坏死因子α（TNF-α）和白细胞介素-6（IL-6），同时增加抗炎因子如白细胞介素-10（IL-10）的水平。Akt信号介导的代谢酶活性的改变与QF显著减少肺脂肪酸积累有关。QF处理后ACLY表达降低，而PPAR表达升高，表明这两种酶是Akt信号的下游靶点，分别控制脂肪酸合成（FAS）和脂肪酸氧化（FAO）。脂肪酸代谢的重编程导致巨噬细胞从M1向M2极化，iNOS表达降低，Arg-1表达升高。此外，Akt激动剂（SC-79）的应用减少QF的抗炎作用，增加FAS和减少巨噬细胞polarization.CONCLUSIONS：QF抑制Akt介导的FAS和极化M1至M2巨噬细胞，导致抗炎的影响。
ETHNOPHARMACOLOGICAL RELEVANCE: Respiratory syncytial virus (RSV) is a common pathogen that causes lower respiratory tract disease in infants and the elderly, and no vaccination is presently available. Qingfei oral liquid (QF), a traditional Chinese medicine formula, has been shown in clinic to have anti-inflammatory properties.AIM OF THE STUDY: The present study investigated whether QF can suppress RSV-induced lung inflammation in mice models via fatty acid-dependent macrophage polarization.MATERIAL AND METHODS: BALB/c mice were given a low, medium, or high dose of QF intragastrically for four consecutive days following RSV infection. The lung inflammatory status was assessed using H&E staining and cytokine assays. The active components of QF and fatty acid metabolism were analyzed using ultra-high-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS). A lipid metabolism-related pathway was found through network pharmacology and molecular docking investigations. Western blotting assays were used to determine the levels of ATP-citrate lyase (ACLY), peroxisome proliferation-activated receptor alpha (PPAR), Akt protein kinase B and its phosphorylated form in Akt signaling. Flow cytometry was used to quantify the number of macrophage subtypes (M1/M2), and immunohistochemistry was used to examine the expression of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1).RESULTS: In the lung tissues of RSV-infected mice, QF suppressed the transcription of pro-inflammatory proteins such as interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6), while increasing the level of anti-inflammatory factors such as interleukin-10 (IL-10). The alterations in metabolic enzyme activity mediated by Akt signaling were linked to QF's significant reduction in lung fatty acid accumulation. Lower ACLY expression and higher PPAR expression were found after QF treatment, showing that these two enzymes were downstream targets of Akt signaling, controlling fatty acid synthesis (FAS) and fatty acid oxidation (FAO), respectively. The reprogramming of fatty acid metabolism resulted in the polarization of macrophages from M1 to M2, with lower expression of iNOS and higher expression of Arg-1. Additionally, application of an Akt agonist (SC-79) reduced QF's anti-inflammatory effects by increasing FAS and decreasing macrophage polarization.CONCLUSIONS: QF inhibited Akt-mediated FAS and polarized M1 to M2 macrophages, resulting in an anti-inflammatory impact.