Nitric oxide-dependent down-regulation of angiotensin II type 2 receptors during experimental sepsis

Objective: The systemic renin-angiotensin system is highly activated during septic shock. This has focused interest in regulation of the adrenal angiotensin II type 2 receptor (AT(2)) as the target thought to mediate angiotensin II-induced adrenal catecholamine release during experimental sepsis in vivo. In addition, the influence of typical endogenous mediators of sepsis, such as proinflammatory cytokines and nitric oxide, on AT(2) receptor expression should be investigated in vitro. Design: Prospective animal trial followed by a controlled cell culture study. Setting: Laboratory of the Department of Anesthesiology. Subjects: Male Sprague-Dawley rats weighing 200-250 g, PC12 cell line. Interventions: Rats were injected with lipopolysaccharide to stimulate Gram-negative sepsis or lipoteichoic acid to stimulate Gram-positive sepsis. AT(2) receptor expression, abundance of the proinflammatory cytokines (interteukin-1 beta, tumor necrosis factor-alpha, and interferon-gamma), and nitric oxide synthase If expression have been determined in the adrenal gland. Rat adrenal pheochromocytoma cells were incubated with these cytokines or with the nitric oxide donors sodium nitroprusside or S-nitroso-N-acetylpenicillamine to investigate the regulation of AT(2) receptors during severe inflammation on a cellular level. Measurements and Main Results: In the adrenal gland, AT(2) receptor expression was down-regulated in both models of sepsis, whereas tissue cytokine concentrations were elevated and nitric oxide synthase II expression was induced. Incubation of PC12 cells with proinflammatory cytokines resulted in a dose-dependent diminished expression of AT(2) receptors, which was mimicked by incubation with nitric oxide donors. Blocking of cytokine-induced nitric oxide synthesis by co-incubation of PC12 cells with N-G-nitro-L-arginine methyl ester prevented down-regulation of AT(2) receptors. Conclusions: These findings show that in our model of sepsis, the expression of AT(2) receptors in the adrenal gland is down-regulated in a nitric oxide-dependent manner. Because AT(2) receptors are thought to be involved in adrenal catecholamine secretion in a stimulatory fashion, the diminished expression of AT(2) receptors could play an important role in the pathogenesis of septic shock via impaired angiotensin II-induced adrenal catecholamine release, despite a strong activation of the systemic renin-angiotensin system.