Inhibition of serine/threonine protein phosphatase PP1 protects cardiomyocytes from tunicamycin-induced apoptosis and I/R through the upregulation of p-eIF2α.

Abstract

The serine/threonine protein phosphatase PP1 mediates the dephosphorylation of phosphorylated eukaryotic translation initiation factor 2 subunit α (p-eIF2α), which is a central regulator of protein synthesis. In the present study, we examined the protective effects of PP1-12 (an inhibitor of the serine/threonine protein phosphatase PP1) against tunicamycin (TM)-induced apoptosis in cultured cardiomyocytes in vitro, as well as in an in vivo model of ischemia/reperfusion (I/R) injury in rat hearts. Neonatal cardiomyocytes cultured from the ventricles of the hearts of 1-day-old Wistar rats were exposed to various concentrations of PP1-12 (0.3, 1 and 3 µmol/l) for 30 min, followed by treatment with TM for 36 h. Cell viability was assessed by adenosine triphosphate (ATP) bioluminescence, and the results revealed that pre-treatment with PP1-12 protected cell viability. Western blot analysis revealed that PP1-12 induced eIF2α phosphorylation and immuncytochemistry indicated that PP1-12 downregulated the expression of C/EBP homologous protein (CHOP), which is related to apoptosis. PP1-12 suppressed cell apoptosis, with maximum protective effects displayed at the concentration of 3 µmol/l. For the in vivo experiments, male Sprague-Dawley rats were randomly divided into 5 groups: i) sham-operated; ii) vehicle (I/R + DMSO); iii) I/R + 1 mg/kg/day PP1-12; iv) I/R + 3 mg/kg/day PP1-12; and v) I/R + 10 mg/kg/day PP1-12. PP1-12 reduced the expression of cleaved caspase-12 and increased the phosphorylation of eIF2α, as revealed by western blot analysis. By calculating the apoptotic index (AI), we found that 10 mg/kg/day PP1-12 exerted the most pronounced anti-apoptotic effect. The infarction area was significantly decreased following treatment with this concentration of PP1-12, as revealed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Taken together, these data suggest that PP1-12 protects cardiomyocytes from TM- and I/R-induced apoptosis, and this effect is achieved at least in part through the inhibition of cell apoptosis and the induction of eIF2α phosphorylation.

• Authors: Chun-Lei Liu, Yun-Yun He, Xin Li, Rui-Jun Li, Kun-Lun He, Li-Li Wang
• Journal: International journal of molecular medicine (Int J Mol Med) Vol. 33 Issue 3 Pg. 499-506 (Mar 2014) ISSN: 1791-244X [Electronic] Greece
• PMID: 24366244 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 4-amino-5-(4-methylphenyl)-7-(tert-butyl)pyrazolo(3,4-d)pyrimidine
• Eukaryotic Initiation Factor-2
• Pyrazoles
• Pyrimidines
• Tunicamycin
• Threonine
• Serine
• Phosphoprotein Phosphatases

Topics

• Animals
• Apoptosis (drug effects)
• Cell Survival (drug effects)
• Eukaryotic Initiation Factor-2 (biosynthesis, metabolism)
• Myocytes, Cardiac (drug effects)
• Phosphoprotein Phosphatases (antagonists & inhibitors, metabolism)
• Phosphorylation (drug effects)
• Pyrazoles (administration & dosage)
• Pyrimidines (administration & dosage)
• Rats
• Reperfusion Injury (drug therapy, metabolism)
• Serine (metabolism)
• Threonine (metabolism)
• Tunicamycin (administration & dosage)