Angiotensin II inhibits interleukin-6 mRNA expression of LPS-stimulated macrophages through down-regulating calcium signaling

BackgroundThe renin-angiotensin system plays a key role in the regulation of blood pressure following hemorrhage and shock. Recent studies also suggest renin-angiotensin system regulates inflammatory mediator production although the amechanism is largely unknown. This purpose of the study was to examine the effect of angiotensin II on macrophage (MØ) IL-6 messenger RNA (mRNA) expression induced by lipopolysaccharide (LPS) and on the alterations in the calcium influx.MethodsJ774A.1 cells, a mouse MØ cell line, were exposed to E. coli LPS (1 or 10 μg/ml) in the presence of angiotensin II (10 nM to 1 μM). IL-6 mRNA expression was determined by the reverse transcription polymerase chain reaction technique. IL-6 protein production was measured by ELISA. To examine the involvement of calcium signaling in IL-6 mRNA expression, MØ were exposed to various calcium agonists and antagonists in the presence of LPS stimulation. Changes of intracellular [Ca2+] by LPS stimulation and angiotensin II treatment were determined by a fura-2 fluorescence ratio method.ResultsLPS stimulation increased MØ IL-6 mRNA expression, which was inhibited by Angiotensin II in a dose-dependent fashion. Both thapsigargin and A23187 augmented the IL-6 mRNA levels induced by LPS stimulation, but only thapsigargin was able to induce IL-6 mRNA directly. TMB-8 but not verapamil inhibited LPS-stimulated MØ IL-6 mRNA. Finally, angiotensin II significantly altered the changes in intracellular [Ca2+] levels induced by LPS stimulation by reducing both the peak and slope of calcium spikes.ConclusionsOur data show that calcium signaling is closely related to IL-6 mRNA expression. Angiotensin II inhibits IL-6 mRNA expression of LPS-stimulated MØ. The inhibitory effects of angiotensin II appear, at least in part, to be mediated through down regulating calcium dependent pathways.