Aquaporin 1 and 5 expression evoked by the β2 adrenoreceptor agonist terbutaline and lipopolysaccharide in mice and in the human monocytic cell line THP-1 is differentially regulated

Abstract

Aquaporin 1 (AQP1) and AQP5 expression may impact on key mechanisms in sepsis. However, it is unclear whether these AQPs are expressed to an equal extent or regulated differentially. Accordingly, we investigated the time-dependent expression of AQP1 and AQP5 following stimulation with lipopolysaccharide (LPS) in cultured human THP-1 cells and in the lungs of mice injected with LPS. Furthermore, we tested the hypothesis that the β2 adrenoreceptor agonist terbutaline or its downstream effector cyclic adenosine monophosphate (cAMP) mitigates LPS-evoked changes of AQP expression. THP-1 cells were stimulated with either LPS (1 μg/mL; serotype O127:B8), 8-Br-cAMP (1 mM), or both, and RNA and protein were extracted at baseline and after 2, 6, and 24 h. C57BL/6 mice that received LPS (20 mg/kg i.p.), terbutaline (2.5 mg/kg), or both were killed 8 h later, and lungs were excised for RNA extraction and lung wet weight determination. Real-time polymerase chain reaction and Western blot analysis show that LPS increased AQP1 (3 h, P < 0.0001) but not AQP5 mRNA and protein expression in THP-1 cells. cAMP increased AQP1 (6 h, P < 0.0001) but not AQP5 mRNA and protein expression. Incubation with both substances accelerated the increase in AQP1 (2 h, P = 0.001) expression, whereas AQP5 expression decreased after 2 h but increased after 24 h (P = 0.0148). In mice lungs, LPS decreased AQP1 (P = 0.0082) but not AQP5 mRNA expression and increased lung wet weight. Terbutaline increased AQP1 mRNA expression twice (P = 0.0005) but not AQP5 mRNA expression. Terbutaline did neither abolish the LPS-induced decrease in AQP1 and AQP5 expression nor increase lung weight. Thus, AQP1 and AQP5 expression is differentially regulated following exposure to LPS, the β2 adrenoreceptor agonist terbutaline, and cAMP. Furthermore, neither terbutaline nor cAMP mitigated the LPS-evoked change of AQP1 and AQP5 expression.