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Pravastatin&[trade] Prevents Venous Shear Stress Dependent Induction of Aquaporin 1 Protein Expression in Human Umbilical Vein Endothelial Cells in vitro
Author(s) -
Crum Raphael J.,
Krane Carissa
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.lb639
Subject(s) - umbilical vein , aquaporin 1 , microbiology and biotechnology , endothelial stem cell , endothelium , immunofluorescence , human umbilical vein endothelial cell , immunocytochemistry , chemistry , biology , biophysics , in vitro , endocrinology , biochemistry , immunology , antibody , mechanical engineering , water channel , engineering , inlet
Background Aquaporins (AQPs) are transmembrane water channels that facilitate osmotically‐driven transmembrane water flux. AQP1 is abundantly expressed in vascular endothelium, where it has been shown to function in endothelial cell migration, wound healing and cell volume regulation in response to mechanical stimuli. AQP1 protein abundance is enhanced in cultured human umbilical vein endothelial cells (HUVECs) in response to fluid flow shear stress. Statins, a pharmacological class of HMG‐CoA reductase inhibitors, have been shown to suppress the early development of vein graft intimal hyperplasia by improving endothelial cell function, likely by affecting endothelial response to arterial shear stress. Statins have also been shown to differentially regulate the expression of specific AQPs by altering their abundance and/or subcellular localization. Therefore, it is hypothesized that AQP1 may function as a component of the mechanosensory complex in endothelial cells, and is subject to shear‐stress induced gene regulation and modulation of expression by statins. Methods HUVECs were cultured in microfluidic chambers under static (0 dynes/cm 2 ) and venous fluid shear stress (6 dynes/cm 2 ) in the presence or absence of 5 μM Pravastatin™. Fluorescent immunocytochemistry was used to detect AQP1 protein expression and subcellular localization. Images were captured using confocal laser scanning microscopy. Mean fluorescent intensity (in arbitrary units) of AQP1 immunofluorescence under each condition was determined using ImageJ software, and normalized to control conditions (time zero, static culture with no Pravastatin™). 25–30 cells from two independent experiments were analyzed for each experimental condition. Results AQP1 immunofluorescence increased 1.3‐fold (p<0.05) after 24 hours, and 1.7‐fold (p<0.05) after 48 hours in cells cultured under venous fluid shear stress as compared to the control. This shear stress‐induced expression was blocked by incubation with Pravastatin™ (p>0.05; control vs. venous shear stress with Pravastatin™). Pravastatin™ had no effect on AQP1 immunofluorescence in static cultures (p>0.05 vs. control). These data indicate that Pravastatin™ prevents shear stress dependent induction of AQP1 in cultured HUVECs. Conclusions The results presented show that the shear stress dependent induction of AQP1 protein expression is prevented by Pravastatin™. These results further demonstrate a possible tissue‐independent function of statin drugs in regulating the expression of AQPs in general and specifically a potential cardioprotective benefit of statins, independent of their cholesterol‐lowering function, related to the regulation of AQP1 in vascular endothelial cells. Support or Funding Information This research was funded by the Schuellein Chair in the Biological Sciences (CMK), University of Dayton Honors Program (RJC), and a 2015 UGSURF Award to RJC.