Mechanisms of Deng-Shi-Qing-Mai-Tang in alleviating PM2.5-Induced lung Injury: Network pharmacology, metabolomics, and molecular Target validation.

ETHNOPHARMACOLOGICAL RELEVANCE: Deng-Shi-Qing-Mai-Tang (DSQMT) is a traditional Chinese herbal formula known for treating inflammatory diseases, particularly those affecting respiratory health. Urban air pollution, especially fine particulate matter (PM2.5), induces lung injury primarily through inflammation and oxidative stress. DSQMT's potential to mitigate PM2.5-induced lung damage makes it a promising ethnopharmacological candidate for pollution-related pulmonary disorders.

AIM OF THE STUDY: This study aims to uncover the therapeutic mechanisms of DSQMT in alleviating PM2.5-induced lung injury, focusing on identifying its active compounds and their molecular targets. By integrating network pharmacology, metabolomics, and experimental validation, we provide a comprehensive understanding of DSQMT's mode of action.

MATERIALS AND

METHODS

We established a rat model of PM2.5-induced lung injury and an in vitro model using PM2.5-treated NR8383 cells. Network pharmacology was applied to predict molecular targets and associated biological pathways affected by DSQMT. Metabolomic profiling identified key metabolic changes, and a pharmacological-metabolomic network was constructed. Molecular docking assessed by binding affinities between DSQMT's active compounds and their targets. The therapeutic effects of DSQMT were evaluated using histological analysis, cytotoxicity assays, and oxidative stress markers, including reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD). Additionally, the roles of Rutin and Morusin, two bioactive compounds, were further validated through RT-qPCR and Western blot to determine their effects on NOS2 and ALOX15 expression.

RESULTS

DSQMT treatment reversed PM2.5-induced metabolic disturbances, restoring homeostasis in key pathways. We identified fifty-one therapeutic targets and fifty metabolites, with NOS2 and ALOX15 emerging as central to DSQMT's protective effects. Molecular docking revealed strong binding between Morusin and NOS2, as well as Rutin and ALOX15. In vitro experiments showed that DSQMT reduced oxidative stress and cytotoxicity, as evidenced by decreased ROS and MDA levels and increased SOD activity. RT-qPCR and Western blot confirmed that Rutin and Morusin modulated NOS2 and ALOX15 expression, validating their contributions to DSQMT's anti-inflammatory and antioxidant effects.

CONCLUSIONS

DSQMT alleviates PM2.5-induced lung injury through metabolic regulation and antioxidant activity. The identification of Rutin and Morusin as key compounds targeting NOS2 and ALOX15 provides mechanistic insights into DSQMT's therapeutic effects. These findings support DSQMT as a potential treatment for PM2.5-related lung injury, highlighting its relevance in managing pollution-induced respiratory diseases.