J Korean Med Sci.  2010 Nov;25(11):1609-1615. 10.3346/jkms.2010.25.11.1609.

Activated Protein C Protects Myocardium Via Activation of Anti-apoptotic Pathways of Survival in Ischemia-reperfused Rat Heart

• Ding JW
• Tong XH
• Yang J
• Liu ZQ
• Zhang Y
• Yang J
• Li S
• Li L

• KMID: 1779231
• DOI: http://doi.org/10.3346/jkms.2010.25.11.1609

Abstract

Activated protein C (APC) is known to be beneficial on ischemia reperfusion injury in myocardium. However, the protection mechanism of APC is not fully understood. The purpose of this study was to investigate the effects and possible mechanisms of APC on myocardial ischemic damage. Artificially ventilated anaesthetized Sprague-Dawley rats were subjected to a 30 min of left anterior descending coronary artery occlusion followed by 2 hr of reperfusion. Rats were randomly divided into four groups; Sham, I/R, APC preconditioning and postconditioning group. Myocardial infarct size, apoptosis index, the phosphorylation of ERK1/2, Bcl-2, Bax and cytochrome c genes and proteins were assessed. In APC-administrated rat hearts, regardless of the timing of administration, infarct size was consistently reduced compared to ischemia/reperfusion (I/R) rats. APC improved the expression of ERK1/2 and anti-apoptotic protein Bcl-2 which were significantly reduced in the I/R rats. APC reduced the expression of pro-apoptotic genes, Bax and cytochrome c. These findings suggest that APC produces cardioprotective effect by preserving the expression of proteins and genes involved in anti-apoptotic pathways, regardless of the timing of administration.

Keyword

Activated Protein C; Cardioprotection; Reperfusion Injury, ERK1/2; bcl-2-Associated X Protein

MeSH Terms

Animals
Apoptosis
Cytochromes c/genetics/metabolism
Hemodynamics/physiology
Male
Mitogen-Activated Protein Kinase 1/metabolism
Mitogen-Activated Protein Kinase 3/metabolism
Myocardial Reperfusion Injury/metabolism/pathology/*prevention & control
Myocardium/*metabolism/pathology
Phosphorylation
Protein C/*therapeutic use
Proto-Oncogene Proteins c-bcl-2/metabolism
Rats
Rats, Sprague-Dawley
Signal Transduction
bcl-2-Associated X Protein/metabolism

Figure

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  Fig. 1 The changes of the hemodynamic parameters before and during myocardial I/R. (A) Left ventricular end-diastolic pressure (LVEDP) during ischemia-reperfusion injury. (B) Left ventricular developed pressure (LVDP). (C) Contractility (+dP/dt) during ischemia-reperfusion injury. dP/dtmin during ischemia-reperfusion injury. (D) Compliance (-dP/dt) during ischemia-reperfusion injury. All values are means±SE (n=6). *P<0.05 for Pre-APC and Post-APC vs the I/R group.

• 

  Fig. 2 The effect of APC on infarct size. Infarct size is expressed as a percentage of the area at risk in rats, which was reduced by APC administration. Mean±SE, n=6. *P<0.01 vs sham group; †P<0.01 vs I/R group.

• 

  Fig. 3 Effect of APC treatment on p-ERK protein in rat heart after 30 min ischemia and 2 h reperfusion. The phosphorylation was quantified by densitometry using image analysis program. Mean±SE, n=6. *P<0.01 vs sham; †P<0.01 vs I/R.

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  Fig. 4 The effect of APC on expressions of Bax, Bcl-2 and cytochrome C mRNA. All values are means±SE (n=6). *P<0.01 vs sham group; †P<0.05 vs I/R group.

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  Fig. 5 Effect of APC on apoptosis in ischemic reperfusion-induced myocardial injury. Detection of apoptosis was done at 2 hr after reperfusion. (A) Sham group, (B) I/R group, (C) Pre-APC group, (D) Post-APC group. (E) Mean apoptotic index was counted in each group (A-D). TUNEL stain ×400 (n=6 animals per group). Arrow indicates TUNEL positive cells. *P<0.05 vs sham group, †P<0.05 vs I/R group.

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