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Updated: Aug 19, 2026
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Published on: December 15, 2023
Exploring the molecular mechanism of danshensu targeting PANoptosis therapy for stroke based on multi omics and
Yuhan Su1, Haifeng Huang1, Hui Qin1
1Department of Neurology, Guangxi Intermational Zhuang Medicine Hospital Affliated to Guangxi University of Chinese Medicine, Nanning City, Guangxi Zhuang Autonomous Region, China.
Background:
One of the main factors aggravating nerve damage following a stroke is the interplay between inflammatory response and cell death. The significance of PANoptosis, a mixed kind of cell death that includes necroptosis, pyroptosis, and apoptosis, in the pathophysiological process of stroke remains unknown.
Aim:
The objective of this work is to systematically identify the core genes of PANoptosis associated with stroke by multi-omics analysis and to investigate the intervention mechanism of Danshensu, the active ingredient of salvianolic acids.
Methods:
We combine the GSE16561 and GSE22255 transcriptome datasets for differential expression analysis, WGCNA, and immune infiltration evaluation; we examine cell heterogeneity and communication networks using the GSE174574 single-cell data; we employ network pharmacology and molecular docking to predict Danshensu's potential targets; finally, we use the BV-2 microglial cell OGD/R model for experimental validation with ZBP1 siRNA knockdown as a functional control.
Results:
A total of 10 PANoptosis core genes (ZBP1, CASP8, TNF, SIRT1, MAPK3, MAPK1, TLR4, NLRP3, MLKL, HMGB1) were identified, mainly enriched in necroptosis, TNF signaling, and cytoplasmic DNA sensing pathways. Immune infiltration showed significant infiltration of neutrophils and M0 macrophages in stroke patients, and single-cell data confirmed that macrophages/microglia are the hub of TNF and MIF signaling networks. Network pharmacology screening identified seven potential targets of Danshensu (TLR4, NLRP3, SIRT1, MAPK1, MAPK3, TNF, CASP8). Molecular docking predicted that Danshensu has binding energies to TNF, MAPK1, and NLRP3 of less than -5 kcal/mol. In vitro experiments confirmed that Danshensu significantly inhibits OGD/R-induced upregulation of ZBP1 and its downstream effectors CASP8, NLRP3, and MLKL at the protein level, reverses SIRT1 downregulation and TLR4/ERK phosphorylation, and exhibits effects similar to ZBP1 siRNA knockdown.
Conclusion:
Our findings suggest that Danshensu may control macrophage inflammatory responses and programmed cell death-related signaling by regulating the PANoptosis-related signaling network, thereby modulating the immunological milieu following a stroke. This study provides novel molecular targets and a theoretical foundation for stroke treatment, although further validation with activation-state markers (e.g., p-MLKL, cleaved CASP8) is warranted.
Insights
This study identifies 10 core PANoptosis genes involved in stroke and shows Danshensu, a natural compound, can regulate these pathways. Danshensu shows promise for treating stroke by modulating inflammation and cell death.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Stroke-induced nerve damage is worsened by inflammation and cell death.
- PANoptosis, a mixed cell death form, is implicated but its role in stroke is unclear.
Purpose of the Study:
- Identify core PANoptosis genes in stroke using multi-omics analysis.
- Investigate the mechanism of Danshensu, a salvianolic acid component, in stroke.
Main Methods:
- Combined transcriptome datasets (GSE16561, GSE22255) for differential expression, WGCNA, and immune infiltration.
- Analyzed single-cell data (GSE174574) for cell heterogeneity and communication.
- Used network pharmacology and molecular docking to predict Danshensu targets.
- Validated findings using a BV-2 microglial OGD/R model and ZBP1 siRNA.
Main Results:
- Identified 10 core PANoptosis genes (ZBP1, CASP8, TNF, SIRT1, MAPK3, MAPK1, TLR4, NLRP3, MLKL, HMGB1) enriched in necroptosis and TNF signaling.
- Found significant neutrophil and M0 macrophage infiltration in stroke; macrophages/microglia are key in TNF/MIF signaling.
- Network pharmacology identified 7 Danshensu targets (TLR4, NLRP3, SIRT1, MAPK1, MAPK3, TNF, CASP8); molecular docking confirmed binding to TNF, MAPK1, NLRP3.
- In vitro studies showed Danshensu inhibits OGD/R-induced ZBP1, CASP8, NLRP3, MLKL, and reverses SIRT1 downregulation and TLR4/ERK phosphorylation.
Conclusions:
- Danshensu may modulate stroke-induced inflammation and cell death by regulating PANoptosis signaling.
- Provides potential therapeutic targets and a theoretical basis for stroke treatment.
- Further validation with specific activation markers is recommended.