Alteration of PKC, PKC theta, NF-kappa B and AP-1 in Ischemic-reperfused Soleus Muscle of Rat with Ischemic Preconditioning

Kim, Tai Seung; Kim, Joo Hak; Kim, Kang Wook

Abstract

PURPOSE
This study was designed to examine the expression patterns of PKC, PKC theta which is a house-keeping member of PKC family in the skeletal muscle, and two redox sensitive transcription factors, NF-kappa B and AP-1.
MATERIALS AND METHODS
Male Sprague-Dawley rats, 9, 30 and 65 weeks old, were divided into 4 groups, such as normal controls, ischemic preconditioning controls, subjects receiving 4 hours of ischemia, and subjects receiving 4 hours of ischemia with ischemic preconditioning. Each group was divided into 5 subgroups based on reperfusion time. For ischemic preconditioning, left common iliac artery was occluded three times for 5 minutes of ischemia followed by 5 minutes of reperfusion using rodent vascular clamps. Ischemia was induced by occluding the same artery for 4 hours. The soleus muscles were removed at hours 0, 1, 3, 6, and 24 after the onset of reperfusion. Muscle samples were embedded in paraffin and 6 micrometer sections were made. The expression levels of PKC, PKC theta, NF-kappa B, and AP-1 were examined using immunohistochemical methods.
RESULTS
Treatment of ischemia for 4 hours followed by reperfusion, resulted in the decrease in PKC, PKC theta, NF-kappa B, and AP-1 in 9 week-old rats, but consistant increase in 30 week-old rats. These factors were highly enhanced in the muscle from 65 week-old rats receiving 3 hours of reperfusion, followed by declined expression. And ischemic preconditioning reduced the expression variability.
CONCLUSION
Ischemic precoditioning reduces the expression variabilities that activated by reperfusion after ischemia, such as PKC, PKC theta, NF-kappa B, and AP-1. But we couldn't predict the tissue damage by the patterns of expression of those factors.