Role of angiotensin II type 1 receptor in angiotensin II-induced cytokine production in macrophages.

ABSTRACT

This study was aimed to investigate the regulatory effects of angiotensin II (Ang II) on the expression of Ang II type 1 (AT1) receptor and to characterize the mechanism underlying AT1 receptor promotion of inflammatory cytokines and reactive oxygen species (ROS). The RAW 264.7 cells were stimulated with various concentrations of Ang II for different times. The mRNA and protein expressions of AT1 receptor in the cells were determined by reverse transcriptase-polymerase chain reaction and Western blot analysis, respectively. The macrophages secretion of proinflammatory cytokines tumor necrosis factor-α, interleukin-1ß (IL-1ß), IL-6, and anti-inflammatory cytokine IL-10 and generation of ROS (by flow cytometry) were also examined. Nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) activities were determined by electrophoretic mobility shift assay. Ang II resulted in an upregulation of AT1 receptor expression in dose- and time-dependent manners in macrophages. Ang II not only induced the production of tumor necrosis factor-α, IL-1ß, IL-6, and IL-10 but also increased the release of ROS. Further, both NF-κB and AP-1 were activated after stimulation with Ang II. However, these events were all abolished by preincubation with ZD7155, a selective competitive antagonist for the AT1 receptor. These results suggest that the AT1 receptor plays an important role in Ang II-induced cytokines production and ROS release via NF-κB and AP-1 pathways in macrophages.