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Cholesterol Enrichment Impairs Capacitative Calcium Entry, eNOS Phosphorylation & Shear Stress-Induced NO Production

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Abstract

Endothelial dysfunction, characterized by decreased production or availability of nitric oxide (NO), is widely believed to be the hallmark of early-stage atherosclerosis. In addition, hypercholesterolemia is considered a major risk factor for development of atherosclerosis and is associated with impaired flow-induced dilation. However, the mechanism by which elevated cholesterol levels leads to decreased production of NO is unclear. NO is released in response to shear stress and agonist-evoked changes in intracellular calcium. Although calcium signaling is complex, we have previously shown that NO production by endothelial nitric oxide synthase (eNOS) is preferentially activated by calcium influx via store-operated channels. We hypothesized that cholesterol enrichment altered this signaling pathway (known as capacitive calcium entry; CCE) ultimately leading to decreased NO. Our results show that cholesterol enrichment abolished ATP-induced eNOS phosphorylation and attenuated the calcium response by the preferential inhibition of CCE. Furthermore, cholesterol enrichment also inhibited shear stress-induced NO production and eNOS phosporylation, consistent with our previous results showing a significant role for ATP autocrine stimulation and subsequent activation of CCE in the endothelial flow response.

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Acknowledgments

We would like to acknowledge the following funding sources for this project: NIH/HL068164 (DJ, KAB), NSF/BES0301446 (DJ, KAB), NSF/CBET0730547 (DJ, KAB), NIH U01HL116256 (DJ, KAB, DGB).

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Authors and Affiliations

  1. Department of Pathology & Laboratory Medicine, Lewis Katz School of Medicine at Temple University, 3500N. Broad St., Philadelphia, PA, 19140, USA

    Allison M. Andrews

  2. School of Biomedical Engineering, Science, and Health Systems, Drexel University, 3141 Market St., Philadelphia, PA, 19104, USA

    Tenderano T. Muzorewa, Kelly A. Zaccheo, Donald G. Buerk, Dov Jaron & Kenneth A. Barbee

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  1. Allison M. Andrews
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  2. Tenderano T. Muzorewa
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Correspondence to Allison M. Andrews.

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Ms. Muzorewa, Ms. Zaccheo, and Dr. Buerk have nothing to disclose. Dr. Andrews, Dr. Jaron and Dr. Barbee have a patent 8,828,711 issued.

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No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Associate Editor William H. Guilford oversaw the review of this article.

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Andrews, A.M., Muzorewa, T.T., Zaccheo, K.A. et al. Cholesterol Enrichment Impairs Capacitative Calcium Entry, eNOS Phosphorylation & Shear Stress-Induced NO Production. Cel. Mol. Bioeng. 10, 30–40 (2017). https://doi.org/10.1007/s12195-016-0456-5

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