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Updated: Mar 29, 2026
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
In Vitro and In Vivo Validation of Endothelium-Derived Potential Therapeutics for Myocardial Ischemia/Reperfusion
Qianlong Zhang1,2,3, Yongsheng Liu3, Zhichao Zhao3
1College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China.
Abstract:
Myocardial ischemia/reperfusion (MI/R) injury affects heart attack outcomes. Endothelial cells dysfunction immediately after MI/R, but the key molecules and how to block them remain unclear. We combined single-cell atlas analysis, AI simulation, and experimental single-cell RNA sequencing data from mouse MI/R; we did quality control, cell annotation, hdWGCNA, and differential gene screening to identify endothelial genes. We constructed a protein network with STRING, predicted structure with AlphaFold3, and used AutoDock for molecular docking to find potential drugs. Virtual knockout simulations were used to check gene deletion effects. The compound andrographolide (AG) was tested in in vitro and in vivo MI/R models by measuring cell viability, inflammation, pathway activity, infarct size, and cardiac function. Single-cell analysis showed that S100 calcium binding protein A8 (S100A8) is an important element in vascular inflammation. It promotes inflammation by interacting indirectly with Cluster of differentiation 14 (CD14). Molecular docking showed that AG binds stably to S100A8. In vitro, AG reduced endothelial injury and blocked the IL-17 pathway. In vivo, AG reduced infarct size, improved cardiac function, and lowered S100A8 and IL-17 pathway proteins. Using single-cell analysis, AI, and experiments, we showed that S100A8 is related to MI/R injury. Andrographolide protects microvasculature via the S100A8 pathway, offering a promising treatment approach and new insights into heart injury mechanisms.
Insights
Myocardial ischemia/reperfusion injury involves endothelial cell dysfunction. Researchers identified S100A8 as a key inflammatory factor and found andrographolide effectively targets it, offering a new treatment for heart attack recovery.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Computational Biology
Background:
- Myocardial ischemia/reperfusion (MI/R) injury impairs endothelial cells, impacting heart attack outcomes.
- Key molecular players and therapeutic targets for MI/R-induced endothelial dysfunction remain largely unknown.
Purpose of the Study:
- To identify critical endothelial genes involved in MI/R injury.
- To explore potential therapeutic interventions targeting identified pathways.
Main Methods:
- Integrated single-cell RNA sequencing, AI simulations, and experimental validation in mouse MI/R models.
- Utilized hdWGCNA for gene screening, STRING and AlphaFold3 for protein network and structure prediction, and AutoDock for molecular docking.
- Assessed the efficacy of andrographolide (AG) in vitro and in vivo MI/R models.
Main Results:
- Single-cell analysis identified S100 calcium binding protein A8 (S100A8) as a key mediator of vascular inflammation in MI/R.
- Molecular docking revealed stable binding of andrographolide to S100A8.
- Andrographolide treatment reduced endothelial injury, inhibited the IL-17 pathway, decreased infarct size, and improved cardiac function in MI/R models.
Conclusions:
- S100A8 is implicated in MI/R injury, promoting vascular inflammation.
- Andrographolide demonstrates therapeutic potential by targeting the S100A8 pathway, protecting the microvasculature.
- This study provides novel insights into MI/R mechanisms and a promising treatment strategy.
