Korean Circ J.  2007 Jul;37(7):327-333. 10.4070/kcj.2007.37.7.327.

Protection of Cardiomyocytes from Acute Ischemic Injury by Protein Kinase Cepsilon Expression

• Yoo JN
• Lee SH
• Jang SI
• Kim SO
• Lee KH
• Park MA
• Lim TH
• Kwon JS
• Cho MC
• Kim YD

• KMID: 2227060
• DOI: http://doi.org/10.4070/kcj.2007.37.7.327

Abstract

BACKGROUND AND OBJECTIVES: Ischemic injury is the most common and important cause of myocardial damage. Over past decades, a number of studies have identified a protective mechanism known as ischemic preconditioning, which can block or delay cell death from ischemic injury. Protein kinase C (PKC), especially theepsilonisoform has been proposed as a key factor in the signaling pathway of ischemic preconditioning. However, whether PKCepsilon expression in cardiomyocytes can offer such protection from acute ischemia has not been explored.
MATERIALS AND METHODS
To demonstrate a direct effect of PKCepsilon expression, a lentiviral vector system was established. Using the lentiviral vector, PKCepsilon was introduced to neonatal rat ventricular myocytes (NRVM) cultured under ischemic conditions, and also to adult rat myocardium subject to left coronary artery ligation.
RESULTS
Compared to control, PKCepsilon expression in cultured NRVM under ischemia resulted in preserved cell density and morphology, and a reduction in cell death (77.6+/-12.8% vs 58.1+/-7.2%, p<0.05). In adult rats, the infarcted area after coronary artery ligation was markedly reduced in myocardium injected with PKCepsilon vector compared to control (11.4+/-5.3% vs 20.5+/-11.3%, p<0.01).
CONCLUSION
These results provide direct evidence that PKCepsilon is a central player in protection against cell death from acute ischemia.

Keyword

Myocytes, cardiac; Protein kinase C-epsilon; Lentivirus; Ischemic preconditioning

MeSH Terms

Adult
Animals
Cell Count
Cell Death
Coronary Vessels
Humans
Ischemia
Ischemic Preconditioning
Lentivirus
Ligation
Muscle Cells
Myocardium
Myocytes, Cardiac*
Protein Kinase C
Protein Kinase C-epsilon
Protein Kinases*
Rats
Protein Kinase C
Protein Kinase C-epsilon
Protein Kinases

Figure

• 

  Fig. 1 Microscopic findings of neonatal rat ventricular myocyte (NRVM) cultured under various conditions. Control: NRVM cultured under normal condition, Ischemia: NRVM cultured under ischemic condition, Ischemia+TPA: NRVM cultured under ischemic condition treated with tetradecanoylphorbol 13-acetate (TPA), Ischemia+vector: NRVM culuted under ischemic condition treated with empty vetor, Ischemia+vector+TPA: NRVM cultured under ischemic condition treated with empty vector and TPA, Ischemia+PKC+TPA: NRVM cultured under ischemic condition treated with protein kinase Cε vector and TPA, PKC: protein kinase C.

• 

  Fig. 2 Comparison of cell death of cultured neonatal rat ventricular myocyte (NRVM) under various conditions. Number of cell death was determined by counting cells failing to exclude the trypan blue dye. Control: NRVM cultured under normal condition, Ischemia: NRVM cultured under ischemic condition, I+TPA: NRVM cultured under ischemic condition treated with tetradecanoylphorbol 13-acetate (TPA), I+PKC+TPA: NRVM cultured under ischemic condition treated with protein kinase Cε vector and TPA. *: p<0.05 between ischemia and ischemia+PKC+TPA, PKC: protein kinase C.

• 

  Fig. 3 Representative samples of transverse section of myocardium stained with phthalocyanin blue and triphenyltetrazolium chloride in control and treated rat heart, injected with PKCε lentiviral vector intramuscularly. Unstainted area (white color) represents infarcted myocardium. PKCε: protein kinase Cε.

• 

  Fig. 4 Bar graphs represent area of infarction between control and treated rat heart, injected with PKCε lentiviral vector intramuscularly. Control MI: rat heart injected with control buffer 4 days prior to left coronary artery ligation, PKCε MI: rat heart injected with PKCε vector 4 days prior to left coronary artery ligation. PKCε: protein kinase Cε, MI: myocardial infarction

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