Cardioprotective effects of Xin-Shu-Ning tablets against myocardial ischemia: Involvement of protein kinase B-related signaling in apoptosis and pyroptosis

ETHNOPHARMACOLOGICAL RELEVANCE Xin-Shu-Ning Tablets (XSNT) are traditionally used to promote blood circulation and resolve blood stasis, including for coronary heart disease-associated angina. Their cardioprotective mechanisms and bioactive constituents remain unclear. AIM OF THE STUDY This study aimed to evaluate the cardioprotection of XSNT against myocardial ischemia and to investigate its underlying mechanisms and candidate bioactive constituents. MATERIALS AND METHODS XSNT was evaluated in myocardial infarction (MI) mice using echocardiography, biochemical assays, histopathology, Western blotting, and immunofluorescence. Proteomics and prior network pharmacology guided PI3K/AKT analysis. H9c2 cells underwent 8 h of oxygen-glucose deprivation (OGD), with MK-2206 used to assess AKT involvement. Molecular docking, molecular dynamics simulations, cellular thermal shift assays, and cellular experiments were used to evaluate the candidates. RESULTS XSNT improved cardiac function and reduced myocardial injury in MI mice. Reduced TUNEL staining and changes in Bcl-2/Bax and cleaved caspase-3 supported attenuation of apoptosis, while reductions in inflammasome- and GSDMD-related proteins indicated decreased pyroptosis-related signaling. These effects accompanied increased AKT phosphorylation and reduced NF-κB activation. XSNT produced similar responses in OGD-challenged H9c2 cells, and MK-2206 partially attenuated these effects. Puerarin showed a potential interaction with AKT and reproduced some cellular effects of XSNT, which were also attenuated by MK-2206. CONCLUSION XSNT attenuated myocardial injury and distinct apoptosis- and pyroptosis-related responses, with pharmacological evidence supporting AKT involvement. Puerarin represents a candidate bioactive constituent. This study has several limitations, including the unclear cellular sources of myocardial pyroptosis-related signals, the lack of genetic validation of AKT involvement, and the need for further confirmation in primary cardiomyocytes.