Objective To evaluate the effects of Naizilai granules in relieving cough, resolving phlegm and ameliorating acute pharyngitis, and to preliminarily explore their underlying mechanisms using untargeted metabolomics and transcriptomics.

Methods The antitussive effect was assessed using a concentrated ammonia water-induced mouse cough model, by measuring cough latency and the number of coughs within 2 min, while the expectorant effect was assessed in a phenol red excretion model by measuring the amount of phenol red excreted in the airway. Untargeted metabolomics was performed to identify differential metabolites and related metabolic pathways between the model control group and the high-dose Naizilai granules group. In a separate rat model of acute pharyngitis induced by topical ammonia application, pharyngeal mucosal pathology was examined via HE staining. The expression of tumor necrosis factor-α (TNF-α) and cyclooxygenase-2 (COX-2) was determined by immunohistochemistry, and inflammation-related signaling pathways were further investigated using whole-blood transcriptomic analysis.

Results Naizilai granules significantly prolonged cough latency and reduced cough frequency in mice. It also increased phenol red secretion from the airways. Metabolomics analysis identified six differential metabolites associated with the expectorant effect, which were primarily enriched in arachidonic acid metabolism, autophagy, and glycerophospholipid metabolism pathways. In rats with acute pharyngitis, Naizilai granules ameliorated inflammatory responses, reduced mucosal thickening, and decreased inflammatory cell infiltration. Immunohistochemical analysis further demonstrated that the granules downregulated the expression of TNF-α and COX-2 in pharyngeal tissues. Whole-blood transcriptomic analysis further revealed that its mechanism of action may be associated with the TNF signaling pathway, Th17 cell differentiation, and T cell activation.

Conclusion Naizilai granules demonstrate good antitussive and expectorant effects, and can effectively alleviate ammonia-induced acute pharyngitis in rats

1. HanX, SunC, DingH, et al. Integration of transcriptomics and metabolomics reveals the mechanism of Glycyrrhizae Radix Et Rhizoma extract inhibiting CCL5 in the treatment of acute pharyngitis[J]. Phytomedicine, 2025, 137: 156360. DOI: 10.1016/j.phymed.2024.156360.

2. XuM, HuTY, LiDC, et al. Yan-Hou-Qing formula attenuates ammonia-induced acute pharyngitis in rats via inhibition of NF-κB and COX-2[J]. BMC Complement Med Ther, 2020, 20(1): 280. DOI: 10.1186/s12906-020-03077-1.

3. 李雪, 刘丹丹, 苏小娟, 等. 利咽止咳汤合养阴清肺汤治疗小儿急性咽炎所致难治性咳嗽疗效观察[J]. 现代中西医结合杂志, 2024, 33(11): 1559-1562.LiX, LiuDD, SuXJ, et al. Clinical efficacy observation of Liyan Zhike decoction combined with Yangyin Qingfei decoction in the treatment of refractory cough caused by acute pharyngitis in children[J]. Modern Journal of Integrated Traditional Chinese and Western Medicine, 2024, 33(11): 1559-1562. DOI: 10.3969/j.issn.1008-8849.2024.11.018.

4. PellegrinoR, TimitilliE, VergaMC, et al. Acute pharyngitis in children and adults: descriptive comparison of current recommendations from national and international guidelines and future perspectives[J]. Eur J Pediatr, 2023, 182(12): 5259-5273. DOI: 10.1007/s00431-023-05211-w.

5. SpinksA, GlasziouPP, Del MarCB. Antibiotics for treatment of sore throat in children and adults[J]. Cochrane Database Syst Rev, 2021, 12(12): CD000023. DOI: 10.1002/14651858.CD000023.pub5.

6. 吴涛. 维药乃孜来颗粒制备工艺、质量标准及药理作用研究[D]. 北京: 中国科学院大学, 2017. https://d.wanfangdata.com.cn/thesis/Ch1UaGVzaXNOZXdTb2xyOVMyMDI2MDkxNjA5NDMzMhIIWTMyNTQ5OTEaCGl6bXJqeHY0.

7. 杨洪军, 李贤煜, 陈鹏, 等. 中药复方的现代基础研究进展述评附视频[J]. 中国科学基金, 2024, 38(3): 387-395.YangHJ, LiXY, ChenP, et al. Review on the progress of modern basic research on traditional Chinese medicine compound prescriptions[J]. Bulletin of the National Natural Science Foundation of China, 2024, 38(3): 387-395. DOI: 10.16262/j.cnki.1000-8217.20240416.001.

8. ChenC, WangJ, PanD, et al. Applications of multi-omics analysis in human diseases[J]. MedComm, 2023, 4(4): e315. DOI: 10.1002/mco2.315.

9. ZhangW, MulatiaS, ZhangM, et al. Esculetin inhibited fever, pain, and inflammatory responses via binding to HSC70[J]. J Ethnopharmacol, 2025, 350: 120022. DOI: 10.1016/j.jep.2025.120022.

10. 王新红, 张迟, 周丽, 等. 基于代谢组学的桔梗总皂苷镇咳祛痰作用机制研究[J]. 药学学报, 2022, 57(3): 757-765.WangXH, ZhangC, ZhouL, et al. The antitussive and expectorant mechanism of Platycodon total saponins based on metabonomics[J]. Acta Pharmaceutica Sinica, 2022, 57(3): 757-765. DOI: 10.16438/j.0513-4870.2021-0719.

11. ZhangJ, XinH, WangW, et al. Investigating the modulatory effects of lactoferrin on depressed rats through 16S rDNA gene sequencing and LC-MS metabolomics analysis[J]. Sci Rep, 2024, 14: 22111. DOI: 10.1038/s41598-024-72793-2.

12. LoveMI, HuberW, AndersS. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J]. Genome Bio, 2014, 15: 550. DOI: 10.1186/s13059-014-0550-8.

13. ZhangY, LiuY, SunJ, et al. Arachidonic acid metabolism in health and disease[J]. MedComm, 2023, 4(5): e363. DOI: 10.1002/mco2.363.

14. DongY, LiuY, TangJ, et al. Zhisou powder displays therapeutic effect on chronic bronchitis through inhibiting PI3K/akt/HIF-1α/VEGFA signaling pathway and reprograming metabolic pathway of arachidonic acid[J]. J Ethnopharmacol, 2024, 319(Pt 1): 117110. DOI: 10.1016/j.jep.2023.117110.

15. 宋海津, 宋琦, 窦宝凯, 等. 多组学联合解析桂枝汤调节脂质代谢表征辛温效应的分子机制附视频[J]. 药学前沿, 2026, 30(1): 30-41.SongHJ, SongQ, DouBK, et al. Exploration of the molecular mechanism of Guizhi Tang's regulation on liver lipid metabolism based on multi-omics analysis[J]. Frontiers in Pharmaceutical Sciences, 2026, 30(1): 30-41. DOI: 10.12173/j.issn.2097-4922.202511020.

16. DuanYW, ChenSX, LiQY, et al. Neuroimmune mechanisms underlying neuropathic pain: the potential role of TNF-α-necroptosis pathway[J]. Int J Mol Sci, 2022, 23(13): 7191. DOI: 10.3390/ijms23137191.

17. 康瑜, 张亚杰, 沈晓丽, 等. 神香草提取物抗炎作用机制研究[J]. 中药材, 2022, 45(1): 187-193.KangY, ZhangYJ, ShenXL, et al. Anti-inflammatory mechanism of extract from the aboveground parts of Hyssopus cuspidatus[J]. Journal of Chinese Medicinal Materials, 2022, 45(1): 187-193. DOI: 10.13863/j.issn1001-4454.2022.01.033.

18. 邢天骄, 彭凌峰, 马丹阳, 等. 新疆一枝蒿不同部位水提物的体外抗炎效果[J]. 中国兽医杂志, 2024, 60(2): 112-119. [XingTJ, PengLF, MaDY, et al. In vitro anti-inflammatory effect of water extracts from different parts of Artemisia rupestris L.[J]. Chinese Journal of Veterinary Medicine, 2024, 60(2): 112-119.https://d.wanfangdata.com.cn/periodical/CiBQZXJpb2RpY2FsQ0hJU29scjkyMDI2MDkxNTE0MjIxORIPemdzeXp6MjAyNDAyMDE3GghzNHlqNzVuaw%3D%3D.