Effect of lipopolysaccharide on angiotensin II type 1 receptor expression and function in human pulmonary microvascular endothelial cells

Abstract

Lipopolysaccharides (LPSs) are an important initiation factor in acute respiratory distress syndrome. The aim of the present study was to investigate the effect of LPSs on the regulation of angiotensin II (Ang II) receptors in human pulmonary microvascular endothelial cells (HPMECs). HPMECs were treated with 0, 50, 100 or 200 ng/ml LPS and incubated for 4, 8, 12 or 16 h. The expression of Ang II type 1 receptor (AT1R) and Ang II type 2 receptor (AT2R) was determined using reverse transcription-polymerase chain reaction and western blot analysis. The affinity to Ang II was measured using a radioligand binding assay. No AT2R expression was detected with or without LPS administration in HPMECs, and LPS treatment increased the expression level of AT1R. A time-dependent increase of AT1R transcription was observed in the 50 ng/ml group, while in the 100 and 200 ng/ml groups, the AT1R mRNA levels reached peak values at 4 h and remained unchanged. The protein level of AT1R increased significantly in a dose-dependent manner for each incubation time period. A time-dependent increase in the protein level was observed in the 50 and 100 ng/ml groups. As for the 200 ng/ml group, the level of AT1R reached a peak value at 8 h. Maximal binding (Bmax) significantly increased following LPS treatment and Bmax of the 50 ng/ml group exhibited a time-dependent increase. The Bmax of the 100 and 200 ng/ml groups reached peak values at 12 and 8 h, respectively, and decreased thereafter. The dissociation constant remained unchanged following LPS treatment. Thus, treatment with LPS increased AT1R expression and its Bmax to Ang II in HPMECs, however, did not alter the affinity of AT1R to Ang II.