Objective To investigate the mechanisms by which Compound Danshen Dropping Pills (复方丹参滴丸) ameliorate myocardial ischemia-reperfusion injury (MIRI) in mice using single-cell and spatial transcriptomics. Methods C57BL/6 mice were randomly divided into sham group, model group, Compound Danshen Dropping Pills low-, medium-, and high-dose (61.43, 122.85, 245.70 mg/kg) groups, with 15 mice in each group. Mice were continuously given drug intervention for 7 d, and a MIRI model was constructed using left anterior descending artery ligation reperfusion method 30 min after the last administration. Cardiac function was assessed by echocardiography 24 h after surgery, and myocardial histopathology was evaluated by hematoxylin-eosin (HE) and Masson staining. Hearts from sham group, model group, and Compound Danshen Dropping Pills medium-dose group were subjected to single-cell and spatial transcriptomic analyses. Single cell transcriptomics was used for cell type identification, cardiomyocyte subpopulation typing, differential gene expression analysis, pseudo temporal analysis, differential gene expression analysis and enrichment analysis. Spatial transcriptomics analysis was used to identify the spatial transcriptomic features of MIRI and the characteristic genes of injury boundary region. Western blotting was performed to detect the expression levels of nuclear receptor subfamily 3 group C member 1 (NR3C1), pyruvate dehydrogenase kinase 4 (PDK4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) in myocardial tissue of mice. Results Compared with sham group, the left ventricular ejection fraction (LVEF) of mice in model group was decreased (P < 0.05), pathology showed significant necrosis of myocardial cells and increased collagen fibers. Compared with model group, Compound Danshen Dropping Pills significantly improved the cardiac function of MIRI mice (P < 0.05), reduced myocardial injury and decreased collagen fibers. The results of single-cell transcriptomics indicated that Compound Danshen Dropping Pills could increase the proportion of myocardial cells and reduce the proportion of fibroblasts and inflammatory cells. Spatial transcriptomics revealed key genes involved in the spatial transcriptomic characteristics and damage boundary regions of MIRI. Western blotting results showed that compared with sham group, the expression levels of NR3C1, PDK4 and ACSL4 proteins in myocardial tissue of mice in model group mice were significantly increased (P < 0.05), while the expression level of GPX4 protein was significantly decreased (P < 0.05). Compared with model group, the expression levels of NR3C1, PDK4 and ACSL4 proteins in myocardial tissue of mice in Compound Danshen Dropping Pills group were significantly reduced (P < 0.05), while the expression level of GPX4 protein was significantly increased (P < 0.05). Conclusion Compound Danshen Dropping Pills may alleviate MIRI in mice by modulating myocardial metabolism and attenuating ferroptosis.

Compound Danshen Dropping Pills  /  single-cell transcriptomics  /  spatial transcriptomics  /  myocardial ischemia-reperfusion injury  /  ferroptosis