Exploring the Mechanism of Therapeutic Effects of Polydatin in Lung Ischemia-Reperfusion Injury by Network Pharmacology and Experiment Validation

Congcong Zhang, Yixin Wang, Haidong Tang, Zhiyi Wu, Shu Jia, Jingyi Wu, Xiaomin Yao Department of Pharmacy, Zhejiang Pharmaceutical University, Ningbo, People’s Republic of ChinaCorrespondence: Xiaomin Yao, Department of Pharmacy, Zhejiang Pharmaceutical University, No. 888, Yinxian Avenue, Yinzhou District, Ningbo, Zhejiang Province, People’s Republic of China, Email [email protected]: To elucidate the therapeutic mechanism of polydatin against lung ischemia-reperfusion injury (LIRI), this study adopted an integrated strategy combining network pharmacology with experimental verification.Methods: Potential targets of polydatin were retrieved from TCMSP, PubChem, SwissTargetPrediction, and Herb. LIRI-related targets were were collected from GeneCards, OMIM, and TTD. Venn analysis was used to identify common targets. A protein–protein interaction (PPI) network was constructed using STRING and analyzed with Cytoscape (CytoNCA plugin).Enrichment analyses (GO/KEGG) were performed to identify key pathways. For experimental validation, an in vitro LIRI model was established in human alveolar epithelial A549 cells undergoing hypoxia/reoxygenation (H/R). The protective effects and associated mechanisms of polydatin were then evaluated through multiple assays, including CCK-8 assay, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), ELISA, TUNEL staining, and reactive oxygen species (ROS) detection. Meanwhile, the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were quantified.Results: Network pharmacology identified 199 potential common targets, with top 10 core ones including AKT1, IL6, TNF, ALB, TP53, INS, IL1B, STAT3, EGFR, and BCL2. GO/KEGG analyses indicated polydatin’s protective effects may relate to inflammatory response modulation, apoptotic signaling regulation, and NF-κB pathway involvement. Experimentally, polydatin enhanced H/R-injured A549 cell viability, reduced apoptosis, increased SOD activity, and decreased MDA, ROS, and inflammatory cytokines (IL-6, IL-1β, TNF-α). It also upregulated AKT1 and ALB mRNA and downregulated TP53 mRNA.Conclusion: Collectively, our results indicate that polydatin alleviates LIRI by attenuating oxidative stress, inflammation, and apoptosis, likely via multi-target and multi-pathway mechanisms centered on key hubs such as AKT1, IL6, TNF, ALB, TP53, and the NF-κB pathway. This study provides a theoretical and experimental basis for further exploration of polydatin as a potential LIRI treatment. Keywords: polydatin, lung ischemia-reperfusion injury, network pharmacology, oxidative stress, inflammation, A549 cells