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Sijunzi Decoction Reverses Metabolic Adaptation and Induces Ferroptosis in Cisplatin-Resistant Non-small Cell Lung
Wen-Jun Liu1,2, Han-Yu Dong3, Chun-Ying Liu4
1Key Laboratory of Ministry of Education for Traditional Chinese Medicine (TCM) Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang, 110847, China.
Objective:
To investigate the mechanistic role of Sijunzi decoction (SJZD) in overcoming chemoresistance through the suppression of adaptive metabolic responses in non-small cell lung cancer (NSCLC).
Methods:
Chemical profiling of SJZD-derived components in systemic circulation was conducted using liquid chromatography-tandem mass spectrometry (LC‒MS/MS) in Sprague-Dawley rats. Multiomics integration and network pharmacology were employed to identify convergent targets shared by the bioactive constituents of SJZD and genes associated with cisplatin resistance. In vitro functional assessments using cisplatin-resistant human lung adenocarcinoma (A549/DDP) cells included the following: quantification of cell viability via Cell Counting Kit-8 (CCK-8) assays; evaluation of mitochondrial bioenergetics through targeted metabolomic profiling; and ultrastructural characterization of ferroptotic morphology via transmission electron microscopy (TEM). Cellular redox homeostasis was dynamically monitored using fluorescent probes, including a DCFH-DA probe for reactive oxygen species (ROS) and a C11-BODIPY581/591 probe for lipid peroxidation. siRNA-mediated gene silencing and immunohistochemical analysis were performed to elucidate the functional hierarchy of the p62/Keap1/nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant axis. Complementary in vivo validation was performed using BALB/c nude mice bearing A549/DDP xenografts, with longitudinal monitoring of tumor progression under SJZD treatment regimens.
Results:
Untargeted metabolomics of SJZD-medicated serum revealed 392 differentially abundant metabolites, with pathway enrichment revealing significant dysregulation of glutamine metabolism. Structural validation confirmed 55 bioactive components of SJZD in serum, including glycyrrhizin, ginsenoside Ro, liquiritigenin, and atractylenolide I. Integration of these components with disease targets yielded 355 overlapping genes associated with both SJZD activity and cisplatin-resistant NSCLC, with significant enrichment in oxidative stress response pathways. Experimental assays confirmed that SJZD induced ferroptosis in cisplatin-resistant A549/DDP cells, as evidenced by disrupted iron homeostasis, lipid peroxidation, and characteristic mitochondrial damage. These effects and subsequent cell death were specifically abrogated by the ferroptosis inhibitor ferrostatin-1 (Fer-1) but not by apoptosis inhibition, confirming that ferroptosis is the primary mechanism of cell death. Mechanistically, the inhibition of p62/Keap1/Nrf2 signaling was involved in the modulation of SJZD-induced ferroptosis both in vitro and in vivo.
Conclusions:
SJZD counteracts metabolic adaptation through ferroptosis mediated by the inhibition of p62/Keap1/Nrf2 in cisplatin-resistant NSCLC.
Insights
Sijunzi decoction (SJZD) overcomes chemoresistance in non-small cell lung cancer (NSCLC) by inducing ferroptosis. SJZD inhibits the p62/Keap1/Nrf2 pathway, suppressing adaptive metabolic responses and promoting cancer cell death.
Area of Science:
- Oncology
- Pharmacology
- Metabolomics
Background:
- Chemotherapy resistance is a major challenge in non-small cell lung cancer (NSCLC) treatment.
- Adaptive metabolic reprogramming contributes significantly to chemoresistance.
- Sijunzi decoction (SJZD) is a traditional Chinese medicine with potential anti-cancer properties.
Purpose of the Study:
- To elucidate the mechanistic role of SJZD in overcoming chemoresistance in NSCLC.
- To investigate SJZD's effect on adaptive metabolic responses in cisplatin-resistant NSCLC cells.
- To identify the molecular pathways targeted by SJZD.
Main Methods:
- Chemical profiling of SJZD components and multiomics integration.
- In vitro studies using cisplatin-resistant lung cancer cells (A549/DDP) including cell viability, mitochondrial bioenergetics, and ferroptosis assays.
- In vivo studies using A549/DDP xenografts in BALB/c nude mice.
- Investigation of the p62/Keap1/Nrf2 signaling pathway.
Main Results:
- SJZD treatment led to significant metabolic alterations, particularly in glutamine metabolism.
- SJZD induced ferroptosis in cisplatin-resistant NSCLC cells, characterized by disrupted iron homeostasis and mitochondrial damage.
- The p62/Keap1/Nrf2 signaling pathway was identified as a key mediator of SJZD-induced ferroptosis.
- SJZD demonstrated anti-tumor efficacy in vivo.
Conclusions:
- SJZD effectively overcomes cisplatin resistance in NSCLC by inducing ferroptosis.
- Inhibition of the p62/Keap1/Nrf2 pathway is a crucial mechanism underlying SJZD's action.
- SJZD represents a promising therapeutic strategy for chemoresistant NSCLC by targeting metabolic adaptation.
