Abstract Objective: Mycoplasma pneumoniae pneumonia (MPP) is the most common community-acquired pneumonia in children aged 5 and above, with clinical manifestations mainly including fever, cough, and headache, posing a significant threat to children’s health. In recent years, especially after the COVID-19 pandemic, epidemiological surveillance data from many countries around the world have shown a significant increase in the incidence and mortality of MPP, making it an important public health challenge. Although most MPP cases are self-limiting and have a good prognosis, some children do not respond well to standard treatment regimens and are prone to developing refractory MPP (RMPP). Therefore, early intervention in MPP is crucial. Traditional Chinese medicine has certain advantages in the early treatment of MPP. According to the “Diagnosis and Treatment Guidelines for Mycoplasma Pneumoniae Pneumonia in Children (2025 Edition)” issued by the National Health Commission of China, Ma-Xing-Shi-Gan Decoction is recommended for treatment. This study aims to explore the pharmacological material basis and mechanism of action of Ma-Xing-Shi-Gan Decoction in the treatment of MPP using network pharmacology methods, providing a scientific basis for its early intervention and rational application in pediatric clinical practice. Methods: Based on the previous UHPLC-MS analysis results of Ma-Xing-Shi-Gan Decoction (containing Ephedra, Bitter Apricot Kernel, Gypsum, and Glycyrrhiza) by our team (Su et al., 2025), the active components and their corresponding targets of Ma-Xing-Shi-Gan Decoction were obtained from the Herb 2.0 and Swiss Target Prediction databases. MPP-related disease targets were screened from databases such as GeneCards, Comparative Toxicogenomics Database (CTD), and Online Mendelian Inheritance in Man (OMIM). The intersection of these targets was taken to obtain potential therapeutic targets. A protein-protein interaction (PPI) network was constructed using the STRING database, and the CentiScape plugin in Cytoscape 3.10.0 software was used to screen core targets based on topological parameters such as degree, betweenness, and closeness. The Metascape database was used to perform gene ontology (GO) biological function annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the core target set. Results: A total of 240 potential therapeutic targets were obtained by intersecting 1830 active component-related targets and 1102 MPP disease targets of Ma-Xing-Shi-Gan Decoction. The PPI network contained 240 nodes and 6332 edges, with an average degree value of 52.8. Topological analysis with parameters of degree: 105.9749, betweenness: 213.0041, and closeness: 0.0022 identified 51 core targets, including IL-6, STAT3, JUN, and MAPK3. KEGG enrichment analysis showed that the core targets were significantly enriched in immune-inflammatory related pathways (such as NF-kappa B signaling pathway), signal transduction related pathways (such as Cytokine-cytokine receptor interaction), and energy metabolism related pathways (such as Insulin resistance); and were widely associated with infection disease pathways such as Influenza A (virus) and tuberculosis (bacteria). GO analysis indicated that the core targets were mainly involved in 1,338 biological processes (Biological Process, BP), such as cytokine stimulation response and oxidative stress response, located in 67 cellular components (Cellular Component, CC), such as membrane rafts and nuclear envelopes, and had 89 molecular functions (Molecular Function, MF), such as protein kinase activity and kinase binding. Conclusion: Based on network pharmacology analysis, this study systematically revealed the “multi-component - multi-target - multi-pathway” synergistic mechanism of Ma-Xing-Shi-Gan Decoction in treating Mycoplasma pneumoniae pneumonia (MPP) in children. The results showed that the active components in the formula, such as ephedrine, glycyrrhizic acid, and amygdalin, might exert their effects by synergistically acting on core targets such as IL-6, STAT3, and MAPK3, regulating core immune inflammatory pathways such as the NF-κB signaling pathway and T cell receptor signaling pathway, thereby interfering with the key links of excessive inflammation and immune imbalance in the early stage of MPP. In addition, the extensive association of core targets with multiple infectious disease pathways suggests that its mechanism of action lies in regulating the common immune network of the body in response to pathogen infection. GO analysis further suggested that it has the potential to exert anti-inflammatory and antioxidant damage effects. In summary, this study systematically explained the multi-dimensional mechanism of Ma-Xing-Shi-Gan Decoction in early intervention of MPP and prevention of its progression to refractory Mycoplasma pneumoniae pneumonia (RMPP), providing a scientific basis for the clinical application of traditional Chinese medicine in the prevention and treatment of pediatric respiratory tract infections. Key words: Mycoplasma pneumoniae pneumonia; Ma-Xing-Shi-Gan Decoction; Network pharmacology; Refractory Mycoplasma pneumoniae pneumonia; Traditional Chinese Medicine

Shuxuan Li is a graduate student and emerging researcher at the Beijing University of Chinese Medicine. Her work stands at the intersection of traditional knowledge and modern science, where she applies advanced computational methods to decode the complex mechanisms of classic TCM formulas. This innovative approach seeks to provide a robust scientific foundation for their clinical use.