Decoding Potential Mechanism of Cucurbitacin IIa Treatment on Lyme Neuroborreliosis Through Integrating Network Pharmacology-Molecular Docking Technique and Cell Experiment

Introduction: Cucurbitacin IIa (CuIIa), one of the most important active components of Cucurbitaceae plants, has a wide range of pharmacological effects. However, the mechanisms underlying its effects on Lyme neuroborreliosis (LNB) remain unclear. This study aimed to elucidate the potential mechanisms of CuIIa activity against LNB. Methods and Results: Potential CuIIa targets were obtained from the Pharmmapper, Swiss Target Prediction, and Batman-Traditional Chinese medicine databases. LNB-associated genes were obtained from OMIM, GeneCards, and DisGeNET. Disease-drug intersection targets were identified using Venny. Protein-protein interaction (PPI) networks were constructed using STRING. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were done on the Database for Annotation, Visualization, and Integrated Discovery (DAVID). A drug-target-pathway-disease network was constructed and Autodock software was used to verify molecular docking between active ingredients and the core targets. Finally, the key targets were experimentally validated. A total of 574 CuIIa targets and 73 LNB-associated genes were identified, and 13 genes were common between the 2 groups. By constructing a PPI network for key targets, the top 10 core target genes were MMP9, TNF, ALB, CTSG, TGFB1, CCL2, IL4, CRP, CCL3, and CCL5. GO functional enrichment and KEGG pathway analyses identified 118 entries and 110 pathways, respectively. Molecular docking results showed that CuIIa binds to key important targets in the core network with high affinity. Validation analyses of the key targets, CCL2 and CCL5, showed that CuIIa decreased their expression in a concentration-dependent manner. Conclusion: This study revealed the potential mechanism of CuIIa activity against Lyme neuroborreliosis. Our preliminary findings using molecular docking modeling and experimental validation provide a basis for future clinical CuIIa applications.