title: 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.affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Chun-Lei
authors.initials: CL
authors.lastname: Liu
authors.affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Yun-Yun
authors.initials: YY
authors.lastname: He
authors.affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Rui-Jun
authors.initials: RJ
authors.lastname: Li
authors.affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Kun-Lun
authors.initials: KL
authors.lastname: He
authors.affiliation: Beijing Institute of Pharmacology and Toxicology, Beijing 100850, P.R. China.
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015852:Eukaryotic Initiation Factor-2 / Q000096:biosynthesis* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011720:Pyrazoles / Q000008:administration & dosage; D011743:Pyrimidines / Q000008:administration & dosage; D051381:Rats; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism; D014415:Tunicamycin / Q000008:administration & dosage
chemical_list: C108123:4-amino-5-(4-methylphenyl)-7-(tert-butyl)pyrazolo(3,4-d)pyrimidine; D015852:Eukaryotic Initiation Factor-2; D011720:Pyrazoles; D011743:Pyrimidines; D014415:Tunicamycin; D013912:Threonine; D012694:Serine; D010749:Phosphoprotein Phosphatases