Department of Postgraduate, Jiangxi University of Chinese Medicine, Nanchang, 330004, Jiangxi Province, China.
College of Pharmacy, Jiangxi University of Chinese Medicine, Nanchang, 330004, Jiangxi Province, China.
J Ethnopharmacol. 2024 Nov 15;334:118584. doi: 10.1016/j.jep.2024.118584. Epub 2024 Jul 15.
Fuzheng-Qushi decoction (FZQS) is a practical Chinese herbal formula for relieving cough and fever. Therefore, the action and specific molecular mechanism of FZQS in the treatment of lung injury with cough and fever as the main symptoms need to be further investigated.
To elucidate the protective effects of FZQS against lung injury in mice and reveal its potential targets and key biological pathways for the treatment of lung injury based on transcriptomics, microbiomics, and untargeted metabolomics analyses.
Lipopolysaccharide (LPS) was used to induce a mouse model of lung injury, followed by the administration of FZQS. ELISA was used to detect IL-1β, IL-6, IL-17A, IL-4, IL-10, and TNF-α, in mouse lung tissues. Macrophage polarization and neutrophil activation were measured by flow cytometry. RNA sequencing (RNA-seq) was applied to screen for differentially expressed genes (DEGs) in lung tissues. RT-qPCR and Western blot assays were utilized to validate key DEGs and target proteins in lung tissues. 16S rRNA sequencing was employed to characterize the gut microbiota of mice. Metabolites in the gut were analyzed using untargeted metabolomics.
FZQS treatment significantly ameliorated lung histopathological damage, decreased pro-inflammatory cytokine levels, and increased anti-inflammatory cytokine levels. M1 macrophage levels in the peripheral blood decreased, M2 macrophage levels increased, and activated neutrophils were inhibited in mice with LPS-induced lung injury. Importantly, transcriptomic analysis showed that FZQS downregulated macrophage and neutrophil activation and migration and adhesion pathways by reversing 51 DEGs, which was further confirmed by RT-qPCR and Western blot analysis. In addition, FZQS modulated the dysbiosis of the gut microbiota by reversing the abundance of Corynebacterium, Facklamia, Staphylococcus, Paenalcaligenes, Lachnoclostridium, norank_f_Muribaculaceae, and unclassified_f_Lachnospiraceae. Meanwhile, metabolomics analysis revealed that FZQS significantly regulated tryptophan metabolism by reducing the levels of 3-Indoleacetonitrile and 5-Hydroxykynurenine.
FZQS effectively ameliorated LPS-induced lung injury by inhibiting the activation, migration, and adhesion of macrophages and neutrophils and modulating gut microbiota and its metabolites.
扶正祛湿汤（FZQS）是一种缓解咳嗽和发热的实用中药配方。因此，需要进一步研究 FZQS 治疗以咳嗽和发热为主要症状的肺损伤的作用和具体分子机制。
通过转录组学、微生物组学和非靶向代谢组学分析，阐明 FZQS 对 LPS 诱导的肺损伤小鼠的保护作用，并揭示其治疗肺损伤的潜在靶点和关键生物学途径。
采用 LPS 诱导小鼠肺损伤模型，给予 FZQS 治疗。采用 ELISA 法检测小鼠肺组织中白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-17A（IL-17A）、白细胞介素-4（IL-4）、白细胞介素-10（IL-10）和肿瘤坏死因子-α（TNF-α）的水平。采用流式细胞术检测 M1 型和 M2 型巨噬细胞极化及中性粒细胞激活。采用 RNA 测序（RNA-seq）筛选肺组织中差异表达基因（DEGs）。采用 RT-qPCR 和 Western blot 检测肺组织中关键 DEGs 和靶蛋白。采用 16S rRNA 测序分析小鼠肠道菌群。采用非靶向代谢组学分析肠道内代谢物。
FZQS 治疗可显著改善 LPS 诱导的肺损伤小鼠的组织病理学损伤，降低促炎细胞因子水平，增加抗炎细胞因子水平。FZQS 还可降低 LPS 诱导的肺损伤小鼠外周血中 M1 型巨噬细胞水平，增加 M2 型巨噬细胞水平，抑制激活的中性粒细胞。转录组学分析显示，FZQS 通过逆转 51 个 DEGs 下调了巨噬细胞和中性粒细胞的激活、迁移和黏附途径，这一结果进一步通过 RT-qPCR 和 Western blot 得到验证。此外，FZQS 通过逆转 Corynebacterium、Facklamia、Staphylococcus、Paenalcaligenes、Lachnoclostridium、norank_f_Muribaculaceae 和 unclassified_f_Lachnospiraceae 的丰度，调节了肠道菌群的失调。同时，代谢组学分析表明，FZQS 通过降低 3-Indoleacetonitrile 和 5-Hydroxykynurenine 的水平，显著调节色氨酸代谢。
FZQS 可通过抑制巨噬细胞和中性粒细胞的激活、迁移和黏附，以及调节肠道菌群及其代谢物，有效改善 LPS 诱导的肺损伤。
