Transfection of antisense p38 alpha gene ameliorates myocardial cell injury mediated by hypoxia and burn serum.

BACKGROUND

Myocardial damage occurs immediately following severe burns even before significant reduction in blood volume. This phenomenon is called postburn "shock heart" ("cardiac shock"), the pathogenesis of which is unclear. This study was designed to investigate the role of antisense p38 alpha gene transfection in ameliorating hypoxia and burn serum-mediated myocardial cell injury.

METHODS

A model of myocardial cells cultured under hypoxia and with burn serum was established. The cells were divided into control group (group C), the group cultured under hypoxia plus burn serum (group HS), and the group treated with antisense p38 alpha gene transfection (group A-p38 alpha) and cultured under hypoxia plus burn serum. Burn serum was collected from Wistar rats with 40% TBSA III degree burns. Hypoxia was produced using a mixed gas with 1% O(2). Antisense p38 alpha gene recombinants were constructed and expression of p38 alpha kinase, and NF-kappaB subunits p50, p65 and I kappa B alpha in myocardial cells were detected by Western blot. Myocardial viability was determined by tetrazolium colorimetry (MTT). Apoptosis was detected by flow cytometry. Lactate dehydrogenase (LDH) activity in cell culture supernatants was determined. Changes in TNFalpha and IL-1 beta mRNA expression were detected by RT-PCR.

RESULTS

Activation of p38 alpha kinase, expression of NF-kappaB p50, NF-kappaB p65 and I kappa B protein, and TNFalpha and IL-1 beta were downregulated significantly following antisense p38 alpha gene transfection into myocardial cells treated with hypoxia plus burn serum. Myocardial apoptosis and LDH activity in cell culture supernatants decreased markedly and myocardial viability increased significantly in the antisense p38 alpha gene treated group.

CONCLUSIONS

Results demonstrated that transfection of antisense p38 alpha gene diminishes myocardial cell injury mediated by hypoxia and burn serum, suggesting a new target for the prevention and treatment of myocardial damage after burn.