2. Academic Validation
  3. The effect of Yinhuang Hanhua dropping pill on acute pharyngitis via a systematic "Compound-Target-Pathway" analysis

The effect of Yinhuang Hanhua dropping pill on acute pharyngitis via a systematic "Compound-Target-Pathway" analysis

  • Phytomedicine. 2025 Jan 26:139:156423. doi: 10.1016/j.phymed.2025.156423.
Qing Zhao 1,  Xiaohuan Wang 2,  Yang Jin 3,  Tiantai Wu 4,  Ming Zhang 5,  Wanfen Shu 6,  Herong Li 7,  Jian Wen 8,  Yan He 9,  Jiyu Chen 10,  Shuaishuai Chen 11,  Ting Liu 12,  Jianfeng Zhang 13,  Wen Liu 14
Affiliations

Affiliations

  • 1 School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. Electronic address: qingzhao.qz@qq.com.
  • 2 School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. Electronic address: 1025031915@qq.com.
  • 3 State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Provincial Key Laboratory of Pharmaceutics, Guizhou Medical University, Guiyang, 550004, China. Electronic address: 842378390@qq.com.
  • 4 School of Basic Medical Sciences, Guizhou Medical University, Guiyang, 550004, China. Electronic address: 804354026@qq.com.
  • 5 School of Basic Medical Sciences, Guizhou Medical University, Guiyang, 550004, China. Electronic address: 527753516@qq.com.
  • 6 School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. Electronic address: 1124773806@qq.com.
  • 7 School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. Electronic address: 269460903@qq.com.
  • 8 School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. Electronic address: 2077979729@qq.com.
  • 9 Clinical Trials Center, The Affiliated Hospital of Guizhou Medical University, Guiyang 550000, China. Electronic address: Yanhe_submission@163.com.
  • 10 Clinical Trials Center, The Affiliated Hospital of Guizhou Medical University, Guiyang 550000, China. Electronic address: chenjiyu@gmc.edu.cn.
  • 11 State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Provincial Key Laboratory of Pharmaceutics, Guizhou Medical University, Guiyang, 550004, China. Electronic address: 1054331382@qq.com.
  • 12 State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Provincial Key Laboratory of Pharmaceutics, Guizhou Medical University, Guiyang, 550004, China. Electronic address: 1586740@qq.com.
  • 13 Guizhou Valid Pharmaceutical Co., Ltd., Guiyang 550014, China. Electronic address: 286911082@qq.com.
  • 14 School of Pharmacy, Guizhou Medical University, Guiyang 550004, China. Electronic address: liuwen@gmc.edu.cn.
PMID: 39951971 DOI: 10.1016/j.phymed.2025.156423
Abstract

Background: Yinhuang Hanhua dropping pill (YHHHDP) is a Chinese herbal formula that is commonly used in medical practice for managing of acute pharyngitis (AP) and Other upper respiratory tract infections. However, the active components, molecular targets, and mechanisms underlying its therapeutic effects remain poorly understood.

Purpose: The objective of this study was to identify the active components and molecular mechanisms of YHHHDP in the therapy of AP.

Methods: The efficacy of YHHHDP was assessed in an ammonia-induced AP rat model and LPS-stimulated RAW264.7 macrophages based on HE staining, immunofluorescence, ELISA, and qRT-PCR. The active components, molecular targets, and pathways were investigated using integrating compositional analysis, metabolomics, and network pharmacological analysis. Furthermore, the interactions between active components and targets, as well as the key signaling pathways were verified using Western blotting (WB), Bio-layer interferometry(BLI), cellular thermal shift assay (CETSA), etc. RESULTS: YHHHDP significantly ameliorated the pathological damage and inflammatory response in the pharyngeal tissues of AP rats. YHHHDP also inhibited LPS-induced expression of pro-inflammatory factors in RAW264.7 cells. Research involving component analysis, network pharmacology, and metabolomics showed that baicalin, chlorogenic acid, and 10 Other compounds are the active components of YHHHDP used in AP treatment. The core targets of these active ingredients were TNF-α, Akt1, and COX-2. The mechanism of action primarily involved the PI3K-AKT/NF-κB/MAPK signaling pathway. Tryptophan and arachidonic acid metabolism were the primary metabolic pathways. Meanwhile, animal and Cellular verification experiments demonstrated that the active ingredients of YHHHDP can stably bind to key target proteins, thereby regulating key metabolites in tryptophan and arachidonic acid metabolism, inhibiting the expression of key proteins in the PI3K-AKT/NF-κB/MAPK signaling pathway, reducing the release of pro-inflammatory factors, and improving the symptoms of AP.

Conclusion: This study revealed that 10 components, such as baicalin and luteolin, are the active ingredients of YHHHDP in AP treatment. The mechanism of action involves the inhibition of TNF-α, Akt1, and COX-2, thereby exerting therapeutic effects through the regulation of arachidonic acid and tryptophan metabolism and the inhibition of the PI3K-AKT/NF-κB/MAPK signaling pathway. This study provided an invaluable scientific basis for the clinical application and development of YHHHDP.

Keywords

Acute pharyngitis; Arachidonic acid metabolic; PI3K-AKT/NF-κB/MAPK signaling pathways; Tryptophan metabolic; Yinhuang Hanhua dropping Pill.

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