Mechanism of Action and Efficacy of Wu-Hua-Yan-Xiao in the Treatment of Pediatric Acute Pharyngitis Based on Network Pharmacology and Experimental Validation

Xinyun Zhang,1,* Zengyu Zhang,2,* Yingzi Xu,1 Jiawei Luo,1 Zihao Shen,3 Hao Liang,1,4 Yidi Zeng,1,4 Wanghua Liu,1,4 Caixing Zheng,1 Jinxia Li1,4 1Hunan University of Chinese Medicine, Changsha, 410208, People’s Republic of China; 2Research Center for Clinical Medicine, Jinshan Hospital Affiliated to Fudan University, Shanghai, 20000, People’s Republic of China; 3Xiangxing College, Hunan University of Chinese Medicine, Changsha, 410208, People’s Republic of China; 4Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, Changsha, 410208, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jinxia Li; Caixing Zheng, Hunan University of Chinese Medicine, Changsha, 410208, People’s Republic of China, Email [email protected]; [email protected] Relevance: Wu-Hua-Yan-Xiao (WHYX) is an innovative volatile oil formulation derived from the traditional Yinqiao-Mabo decoction, developed for the treatment of pediatric acute pharyngitis.Materials and Methods: Network pharmacology was utilized to identify active components and potential therapeutic targets of WHYX in acute pharyngitis. Compounds in WHYX were characterized using UHPLC-QE-MS. A pediatric acute pharyngitis rat model was established by administering 25% ammonia to the pharyngeal mucosa of young rats. WHYX was delivered via aerosol inhalation at gradient concentrations. Histopathological changes in pharyngeal tissues were evaluated by H&E staining. Serum levels of IL-6, IL-1β, TNF-α, and PGE2 were quantified by ELISA. Expression levels of TNF-α, TP53, IL17A, IL6, and Bcl-2 were assessed by qRT-PCR and Western blotting. Apoptosis was analyzed through immunofluorescence staining for Caspase-3 and TUNEL.Results: Network pharmacology identified 130 active compounds and 600 gene targets, with 194 overlapping drug-disease targets. TP53 signaling emerged as a central regulatory pathway. Compared with the model group, the high-dose WHYX volatile oil group showed marked improvements in pharyngeal pathology, significant reductions in inflammatory cytokines, downregulation of TNF-α, TP53, IL17A, IL6, and Bcl-2 expression, and suppressed apoptosis (P < 0.05). Therapeutic effects were comparable to or exceeded those observed in the positive control group. (P < 0.05).Conclusion: The WHYX formula alleviates inflammation, reduces apoptosis, and protects pharyngeal tissue in young rats with acute pharyngitis. Aerosol inhalation of WHYX presents a direct, effective, and non-invasive strategy for pediatric acute pharyngitis management. Keywords: pediatric acute pharyngitis, traditional Chinese medicine volatile oil, atomization inhalation, network pharmacology, cell apoptosis, anti-inflammatory effects