Fufang Danshen Pill improves mitochondrial homeostasis by regulating the S100a9/TLR4 axis to alleviate myocardial ischemia-reperfusion injury

  • Phytomedicine. 2025 Oct 21:148:157433. doi: 10.1016/j.phymed.2025.157433.
Ruixia Lu  1 ,  Wenhao Sun  1 ,  Wenyong Lin  1 ,  Siyue Zhu  1 ,  Dongyuan Wang  1 ,  Qipeng Jin  1 ,  Dan Wang  1 ,  Zhenchi Sang  2 ,  Xiaolong Wang  3
Affiliations
  • 1. Branch of National Clinical Research Center for Chinese Medicine Cardiology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200120, China; Cardiovascular Research Institute of Traditional Chinese Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200120, China.
  • 2. Department of Cardiology, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200030, China.
  • 3. Branch of National Clinical Research Center for Chinese Medicine Cardiology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200120, China; Cardiovascular Research Institute of Traditional Chinese Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200120, China. Electronic address: [email protected].
PMID: 41166904 DOI: 10.1016/j.phymed.2025.157433
Abstract

Background: S100a9 protein participates in regulating mitochondrial damage during Myocardial ischemia-reperfusion injury(I/R), but its cell type-specific mechanisms remain unclear. Fufang Danshen Pill (FDP), a traditional Chinese medicine for I/R treatment, requires further investigation to elucidate its therapeutic targets.

Purpose: Investigate the macrophage-specific role of S100a9 in I/R pathogenesis, and clarifies the molecular pathways through which FDP improves mitochondrial function by modulating S100a9.

Methods: The murine I/R model was established and subjected to determine the optimal therapeutic dosage of FDP. By integrating Single-cell transcriptomics(scRNA-seq)with spatial transcriptomics, we conducted comprehensive analyses of myocardial tissues from sham, I/R, and FDP groups, including cellular subtyping, intercellular communication analysis, and spatial transcriptomic mapping. Mechanistic validation was performed using Adeno-associated virus serotype 2/9(AAV2/9)-mediated macrophage-specific S100a9 overexpression models to confirm key regulatory targets.

Results: The medium-dose FDP (122.85 mg/kg) exhibited the best efficacy in improving cardiac function. FDP intervention markedly attenuated macrophage-cardiomyocyte cellular crosstalk. In I/R models, cardiomyocytes displayed significant alterations in mitochondrial-associated gene expression, whereas S100a9 emerged as the predominant differentially expressed gene in macrophages. Spatial transcriptomic mapping localized S100a9 predominantly within myocardial injury regions. Functional investigations revealed that AAV2/9-mediated macrophage-specific S100a9 overexpression exacerbated I/R pathological manifestations, including increased infarct size, reduced ejection fraction, elevated serum cardiac troponin levels, TLR4 signaling activation, and mitochondrial dysfunction. Notably, FDP treatment substantially reversed these pathological alterations.

Conclusion: This study demonstrates that macrophage-derived S100a9 mediates mitochondrial damage in cardiomyocytes during I/R injury. FDP effectively maintains mitochondrial functional homeostasis in cardiomyocytes by targeting and inhibiting macrophage S100a9 expression. These findings provide a important scientific basis for developing intervention strategies targeting the S100a9/TLR4 axis.

Keywords
Fufang Danshen Pill; I/R injury; Mitochondrial activity; S100a9; Single-cell transcriptomics sequencing; Spatial transcriptomics sequencing.
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