Bioactive equivalent combinatorial components of Xiao-Xu-Ming decoction inhibit the calmodulin-mediated MLCK/MLC axis to attenuate coronary artery spasm

  • Phytomedicine. 2025 Jul:142:156713. doi: 10.1016/j.phymed.2025.156713.
Bowen Wu  1 ,  Mishan Wu  2 ,  Peiyu Yan  3
Affiliations
  • 1. Faculty of Chinese Medicine, Macau University of Science and Technology, Taipa 999078, Macao, China.
  • 2. Faculty of Chinese Medicine, Macau University of Science and Technology, Taipa 999078, Macao, China; State Key Laboratory of Quality Research in Chinese Medicines, Macau University of Science and Technology, Taipa 999078, Macao, China.
  • 3. Faculty of Chinese Medicine, Macau University of Science and Technology, Taipa 999078, Macao, China; State Key Laboratory of Quality Research in Chinese Medicines, Macau University of Science and Technology, Taipa 999078, Macao, China. Electronic address: [email protected].
PMID: 40349422 DOI: 10.1016/j.phymed.2025.156713
Abstract

Background: Coronary artery spasm (CAS) is a severe pathological disorder with limited treatment options. Inhibiting vascular smooth muscle cell (VSMC) proliferation has emerged as a promising therapeutic strategy for CAS.

Purpose: This study aims to investigate the potential function of Xiao-Xu-Ming decoction (XXMD) in the attenuation of CAS in rats.

Methods: The involvement of bioactive equivalent combinatorial components (BECCs) of XXMD in CAS was predicted using network pharmacological analysis. A CAS rat model was established using pituitrin (Pit), and an Ang II-induced cell model was developed to assess the effects of BECCs on VSMC contraction. The expression of contractile phenotype markers was analyzed using RT-qPCR and Western blotting. Additionally, MTT assay, flow cytometry, Annexin V/PI staining, and Transwell assay were performed to evaluate cell proliferation, cell cycle progression, Apoptosis, and migration. A Collagen gel contraction assay was conducted to assess VSMC contraction.

Results: Network pharmacological analysis suggested that BECCs in XXMD may influence CAS development through the Calmodulin (CaM)-MLCK/MLC axis. The drug-containing XXMD serum significantly inhibited VSMC proliferation and migration by reducing the expression of MLCK, α-SMA, Calponin, and SMHHC. Furthermore, it suppressed VSMC proliferation by downregulating MLCK expression via CaM inhibition. Importantly, BECCs of XXMD promoted anti-inflammatory responses, vasorelaxation, and oxidative stress reduction in Pit-induced CAS rats.

Conclusion: These findings suggest that BECCs in XXMD may counteract CAS by modulating the CaM-mediated MLCK/MLC pathway in VSMCs. This mechanism may offer a foundation for novel and effective therapeutic strategies to treat CAS and prevent associated cardiac events, such as myocardial infarction and sudden cardiac death.

Novelty: This study identifies a novel mechanism by which the "active ingredient cluster" of XXMD alleviates CAS through modulation of the MLCK/MLC signaling pathway via CaM. These insights enhance our understanding of XXMD's therapeutic potential in CAS treatment.

Keywords
Bioactive equivalent combinatorial components; Calmodulin; Coronary artery spasm; MLCK/MLC; Vascular smooth muscle cells; Xiao-Xu-Ming decoction.
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