Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2

Rescuing cells from stress damage emerges a potential therapeutic strategy to combat myocardial infarction. Protocatechuic aldehyde (PCA) is a major phenolic acid in Chinese herb Danshen (Salvia miltiorrhiza root). This study investigated whether PCA regulated nuclear Pyruvate Kinase isoform M2 (PKM2) function to protect cardiomyocytes. In rats subjected to isoprenaline, PCA attenuated heart injury and protected cardiomyocytes from Apoptosis. Through DARTS and CETSA assays, we identified that PCA bound and promoted PKM2 nuclear translocation in cardiomyocytes exposed to oxygen/glucose deprivation (OGD). In the nucleus, PCA increased the binding of PKM2 to β-catenin via preserving PKM2 acetylation, and the complex, in cooperation with T-cell factor 4 (TCF4), was required for transcriptional induction of genes encoding anti-apoptotic proteins, contributing to rescuing cardiomyocyte survival. In addition, PCA ameliorated mitochondrial dysfunction and prevented mitochondrial Apoptosis dependent on PKM2. Consistently, PCA increased the binding of PKM2 to β-catenin, improved heart contractive function, normalized heart structure and attenuated oxidative damage in mice subjected to artery ligation, but the protective effects were lost in Pkm2-deficient heart. Together, we showed that PCA regulated nuclear PKM2 function to rescue cardiomyocyte survival via β-catenin/TCF4 signaling cascade, suggesting the potential of pharmacological intervention of PKM2 shuttle to protect the heart.

Apoptosis; CETSA, cellular thermal shift assay; CK-MB, creatine kinase isoenzyme-MB; DARTS, drug affinity responsive target stability; Heart ischemia; ISO, isoprenaline; LDH, lactate dehydrogenase; Mitochondrial damage; Myocardial infarction; NRVMs, neonatal rat ventricular myocytes; Nuclear translocation; OGD, oxygen and glucose deprivation; PCA, protocatechuic aldehyde; PKM2; PKM2, pyruvate kinase isoform M2; Protocatechuic aldehyde; ROS, reactive oxygen species; TCF4; TCF4, T-cell factor 4; TUNEL, deoxynucleotidyl transferase-mediated dUTP nick end-labeling; shRNA, short hairpin RNA; β-Catenin.