Effects Of Egr-1 Antisense Oligodeoxynucleotide On Egr-1 Expression And Cardiomyocytes Hypoxia-reoxygenation Injury

The research about myocardial ischemia-reperfusion(I/R) injury is an important area in cardiology. Many literatures reported that Egr-1 played a significant role during I/R injury. In the experiment, the effect of antisense oligodeoxynucleotide (AS-ODN) on Egr-1 protein expression, inflammatory injury and cellular function in the model of cultured rat cardiomyocytes were evaluated undergoing hypoxia-reoxygenation(H/R). This article investigated that Egr-1 played an important role in the occurrance and development of cardiomyocytes H/R injury, and AS-ODN could relieve H/R injury.I. Effects of AS-ODN on Egr-1 expression and cardiomyocytes inflammationVentricular myocardial cells from neonatal rats were isolated enzymatically and cultured in M₁₉₉ culture medium for 5 days. Cells were transfected with FITC labeled Egr-1 AS-ODN for 8h.To follow this procedure, cardiomyocytes were washed with PBS, fixed with acetone and mounted by glycerin for taking photographs under fluorescente microscope.Cardiomyocytes were randomly divided into 6 groups: 1. normal(N), 2. hypoxia-reoxygenation(H/R), 3. lipofectamine? 2000(LIP), 4. sense oligodeoxynucleotide (S), 5. scrambled oligodeoxynucleotide(Sc), 6. antisense oligodeoxynucleotide(AS). The cells of 2, 3, 4, 5 and 6 groups were treated with hypoxia for 3 hours and subsequently reoxygenated for 1 hour. TNF- α was assayed from culture medium by ELISA and Egr-1 production in cardiomyocytes was measured by Western blot.