Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation
- 1Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, 030006, China; The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, China
- 2First Clinical Medical College, Shanxi Medical University, Taiyuan, 030000, China; National Health Commission Key Laboratory of Pneumoconiosis, Shanxi Key Laboratory of Respiratory Diseases, Department of Pulmonary and Critical Care Medicine, The First Hospital of Shanxi Medical University, Taiyuan, 030000, China
- 3Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, 030006, China; The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, China; Corresponding author. Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, 030006, China.
Abstract
Pneumoconiosis is a common occupational lung disease with unclear pathogenesis and no effective targeted therapies. Emerging evidence suggests that daphnetin is a promising therapeutic candidate for silicosis, yet its efficacy targets remain poorly characterized. In this study, we propose and implement a multi-layered target identification strategy that integrates transcriptomics, clinical metabolomics, public database mining, an artificial intelligence (AI)-based target prediction model, and tissue-based thermal proteome profiling (Tissue-TPP) technology. This strategy enables stepwise elucidation, from pneumoconiosis-related targets to daphnetin efficacy targets and ultimately to direct targets. Using this approach, we validated nicotinamide adenine dinucleotide phosphate hydrogen (NAD(P)H) quinone oxidoreductase 1 (NQO1) and 5-oxoprolinase (OPLAH) as direct targets of daphnetin, and identified Kelch-like ECH-associated protein 1 (KEAP1), nuclear factor erythroid 2-related factor 2 (NRF2), and arginase 1 (ARG1) as indirect targets. Notably, OPLAH is reported here as a direct target of daphnetin for the first time. Furthermore, the binding of daphnetin to NQO1 and OPLAH helps explain its effects on cellular thiol levels. Using a thiol-responsive probe, we observed that daphnetin significantly increased intracellular thiol levels and reduced silica-induced reactive oxygen species (ROS) accumulation in both A549 and THP-1 cells, thereby contributing to its antioxidant effects. Additionally, overexpression (OE) of OPLAH significantly increased reduced glutathione (GSH) levels and alleviated silica-induced epithelial-mesenchymal transition (EMT). Conversely, OPLAH knockdown partially inhibited the efficacy of daphnetin. Together, these findings clarify the anti-fibrotic mechanism of daphnetin, support its therapeutic potential for silicosis, and provide a practical framework for the target discovery of natural products (NPs).
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BibTeX
@article{wang2026multilayered,
title = {{Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation}},
author = {Ling Wang and Bo-Yan Ma and Sheng-Ping Jiang and Shu-Ting Wei and Xue-Mei Qin and Zhen-Yu Li},
journal = {Journal of Pharmaceutical Analysis},
year = {2026},
month = sep,
doi = {10.1016/j.jpha.2026.101629},
url = {https://doi.org/10.1016/j.jpha.2026.101629},
note = {CC BY},
}
RIS
TY - JOUR
TI - Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation
AU - Ling Wang
AU - Bo-Yan Ma
AU - Sheng-Ping Jiang
AU - Shu-Ting Wei
AU - Xue-Mei Qin
AU - Zhen-Yu Li
PY - 2026
DA - 2026/09/01
JO - Journal of Pharmaceutical Analysis
DO - 10.1016/j.jpha.2026.101629
UR - https://doi.org/10.1016/j.jpha.2026.101629
AB - Pneumoconiosis is a common occupational lung disease with unclear pathogenesis and no effective targeted therapies. Emerging evidence suggests that daphnetin is a promising therapeutic candidate for silicosis, yet its efficacy targets remain poorly characterized. In this study, we propose and implement a multi-layered target identification strategy that integrates transcriptomics, clinical metabolomics, public database mining, an artificial intelligence (AI)-based target prediction model, and tissue-based thermal proteome profiling (Tissue-TPP) technology. This strategy enables stepwise elucidation, from pneumoconiosis-related targets to daphnetin efficacy targets and ultimately to direct targets. Using this approach, we validated nicotinamide adenine dinucleotide phosphate hydrogen (NAD(P)H) quinone oxidoreductase 1 (NQO1) and 5-oxoprolinase (OPLAH) as direct targets of daphnetin, and identified Kelch-like ECH-associated protein 1 (KEAP1), nuclear factor erythroid 2-related factor 2 (NRF2), and arginase 1 (ARG1) as indirect targets. Notably, OPLAH is reported here as a direct target of daphnetin for the first time. Furthermore, the binding of daphnetin to NQO1 and OPLAH helps explain its effects on cellular thiol levels. Using a thiol-responsive probe, we observed that daphnetin significantly increased intracellular thiol levels and reduced silica-induced reactive oxygen species (ROS) accumulation in both A549 and THP-1 cells, thereby contributing to its antioxidant effects. Additionally, overexpression (OE) of OPLAH significantly increased reduced glutathione (GSH) levels and alleviated silica-induced epithelial-mesenchymal transition (EMT). Conversely, OPLAH knockdown partially inhibited the efficacy of daphnetin. Together, these findings clarify the anti-fibrotic mechanism of daphnetin, support its therapeutic potential for silicosis, and provide a practical framework for the target discovery of natural products (NPs).
SN - 2095-1779
SN - 2214-0883
ER -