[Effects of burn patients' serum on IkappaBalpha degradation and NF-kappaB activation in endothelial cells in vitro].
ABSTRACT To investigate the effects of the serum from burned patients (burn serum) on inhibitor of kappaBalpha (IkappaBalpha) degradation and nuclear factor-kappaB (NF-kappaB) activation in the endothelial cells in order to explore the role of burn serum on endothelium activation.
Cultured endothelial cells (ECV-304) were stimulated by the serum from healthy volunteers, burn serum and burn serum with pyrrolidine dithiocarbamate (PDTC), respectively. Activation of endothelial NF-kappaB at 30, 60, 120, 240 min after stimulation was tested by electrophoretic mobility shift assay (EMSA) and the degradation of endothelial IkappaBalpha at these time points determined by Western blotting.
Compared with that in the control group, cytosolic IkappaBalpha degradation in the endothelial cells occurred within 30 min after burn serum stimulation, and peaked at 60 min, followed by gradual recovery in the cytoplasm till reaching the pre-stimulation level at 2 h. Meanwhile, the activity of endothelial NF-kappaB was markedly increased, reaching the peak at 30 to 60 min with subsequent gradual decrease 2 h after stimulation. PDTC could effectively inhibit endothelial IkappaBalpha degradation and activation of NF-kappaB induced by the burn serum.
Burn serum might induce the degradation of IkappaBalpha to activate NF-kappaB, which ultimately leads to cytokine release from the endothelium.