Empagliflozin prevents oxidative stress in human coronary artery endothelial cells via the NHE/PKC/NOX axis
Xiaoling Li,
Mengnan Wang,
Jan-Ole Kalina,
Benedikt Preckel,
Markus W. Hollmann,
Martin Albrecht,
Coert J. Zuurbier,
Nina C. Weber
Affiliations
Xiaoling Li
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands
Mengnan Wang
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands
Jan-Ole Kalina
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands; Department of Anesthesiology and Intensive Care Medicine, Universitätsklinikum Schleswig-Holstein, Campus Kiel, 24105, Kiel, Germany
Benedikt Preckel
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands
Markus W. Hollmann
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands
Martin Albrecht
Department of Anesthesiology and Intensive Care Medicine, Universitätsklinikum Schleswig-Holstein, Campus Kiel, 24105, Kiel, Germany
Coert J. Zuurbier
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands
Nina C. Weber
Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology-L.E.I.C.A, Amsterdam Cardiovascular Science (ACS), Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands; Corresponding author. Amsterdam, University Medical Centers, Location AMC, Department of Anesthesiology - L.E.I.C.A, Amsterdam Cardiovascular Science, Meibergdreef 11, 1105 AZ, Amsterdam, the Netherlands.
Background: Empagliflozin (EMPA) ameliorates reactive oxygen species (ROS) generation in human endothelial cells (ECs) exposed to 10 % stretch, but the underlying mechanisms are still unclear. Pathological stretch is supposed to stimulate protein kinase C (PKC) by increasing intracellular calcium (Ca2+), therefore activating nicotinamide adenine dinucleotide phosphate oxidase (NOX) and promoting ROS production in human ECs. We hypothesized that EMPA inhibits stretch-induced NOX activation and ROS generation through preventing PKC activation. Methods: Human coronary artery endothelial cells (HCAECs) were pre-incubated for 2 h before exposure to cyclic stretch (5 % or 10 %) with either vehicle, EMPA or the PKC inhibitor LY-333531 or PKC siRNA. PKC activity, NOX activity and ROS production were detected after 24 h. Furthermore, the Ca2+ chelator BAPTA-AM, NCX inhibitor ORM-10962 or NCX siRNA, sodium/potassium pump inhibitor ouabain and sodium hydrogen exchanger (NHE) inhibitor cariporide were applied to explore the involvement of the NHE/Na+/NCX/Ca2+ in the ROS inhibitory capacity of EMPA. Results: Compared to 5 % stretch, 10 % significantly increased PKC activity, which was reduced by EMPA and PKC inhibitor LY-333531. EMPA and LY-333531 showed a similar inhibitory capacity on NOX activity and ROS generation induced by 10 % stretch, which was not augmented by combined treatment with both drugs. PKC-β knockdown inhibits the NOX activation induced by Ca2+ and 10 % stretch. BAPTA, pharmacologic or genetic NCX inhibition and cariporide reduced Ca2+ in static HCAECs and prevented the activation of PKC and NOX in 10%-stretched cells. Ouabain increased ROS generation in cells exposed to 5 % stretch. Conclusion: EMPA reduced NOX activity via attenuation of the NHE/Na+/NCX/Ca2+/PKC axis, leading to less ROS generation in HCAECs exposed to 10 % stretch.
