Korean J Physiol Pharmacol.  2008 Oct;12(5):275-280. 10.4196/kjpp.2008.12.5.275.

Kinetic Changes of COX-2 Expression during Reperfusion Period after Ischemic Preconditioning Play a Role in Protection Against Ischemic Damage in Rat Brain

• Kang YJ
• Park MK
• Lee HS
• Choi HC
• Lee KY
• Kim HJ
• Seo HG
• Lee JH
• Chang KC

• KMID: 1486110
• DOI: http://doi.org/10.4196/kjpp.2008.12.5.275

Abstract

A brief ischemic insult induces significant protection against subsequent massive ischemic events. The molecular mechanisms known as preconditioning (PC)-induced ischemic tolerance are not completely understood. We investigated whether kinetic changes of cyclooxygenase (COX)-2 during reperfusion time-periods after PC were related to ischemic tolerance. Rats were given PC by occlusion of middle cerebral artery (MCAO) for 10 min and sacrificed after the indicated time-periods of reperfusion (1, 2, 4, 8, 12, 18 or 24 h). In PC-treated rats, focal ischemia was induced by occlusion of MCA for 24 h and brain infarct volume was then studied to determine whether different reperfusion time influenced the damage. We report that the most significant protection against focal ischemia was obtained in rats with 8 h reperfusion after PC. Administration of indomethacin (10 mg/kg, oral) or rofecoxib (5 mg/kg, oral) 48 h prior to PC counteracted the effect of PC. Immunohistochemical analysis showed that COX-2 and HO-1 protein were induced in PC-treated rat brain, which was significantly inhibited by rofecoxib. Taken together, we concluded that the kinetic changes of COX-2 expression during the reperfusion period after PC might be partly responsible for ischemic tolerance.

Keyword

Ischemic preconditioning; Stroke; Heme oxygenase; Cyclooxygenase

MeSH Terms

Animals
Brain
Heme Oxygenase (Decyclizing)
Indomethacin
Ischemia
Ischemic Preconditioning
Lactones
Middle Cerebral Artery
Prostaglandin-Endoperoxide Synthases
Rats
Reperfusion
Stroke
Sulfones
Heme Oxygenase (Decyclizing)
Indomethacin
Lactones
Prostaglandin-Endoperoxide Synthases
Sulfones

Figure

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  Fig. 1. Time course of COX-2 expression by Western blot analysis in rat brain subjected to MCA occlusion for 10 min. The results were presented as ratio of COX-2/β-actin, in which the amount of constitutively expressed COX-2/β-actin at 0 h was designated as 1. It should be noted that preconditioning increases COX-2 expression which varied depending on the reperfusion.

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  Fig. 2. A representative photograph of neuronal damage between sham-operated (A) and ischemic preconditioned rat brain (B), in which 8 h reperfusion was performed after 10 min MCA occlusion.

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  Fig. 3. Size of brain infarction according to reperfusion time after PC against focal ischemia (24 h, MCAO). (A). Percentile of neuronal damage (%) occurred with different time of reperfusion after ischemic preconditioning against focal ischemia (B).

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  Fig. 4. A representative photograph of neural damage by COX-inhibitors in PC-treated rat brain. A. PC only, B. Indomethacin/PC, C. roferocoxib/PC (A) Percentile of neuronal damage (%) occurred with COX-inhibitors in PC-treated rat (B).

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  Fig. 5. Immunohisotochemical staining of COX-2 and HO-1 in the brain treated with or without rofecoxib in PC-treated rat. (A) COX-2 (B) HO-1.

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