Calciprotein Particles Induce Endothelial Dysfunction by Impairing Endothelial Nitric Oxide Metabolism

Lian Feenstra, Anton G. Kutikhin, Daria K. Shishkova, Hendrik Buikema, Lara Zeper, Arno R. Bourgonje, Guido Krenning, Jan-Luuk Hillebrands

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Abstract

BACKGROUND:
Calciprotein particles (CPPs) are associated with the development of vascular calcifications in chronic kidney disease. The role of endothelial cells (ECs) in this process is unknown. Here, we investigated the interaction of CPPs and ECs, thereby focusing on endothelial nitric oxide metabolism and oxidative stress.

METHODS:
CPPs were generated in calcium- and phosphate-enriched medium. Human umbilical vein endothelial cells were exposed to different concentrations of CPPs (0–100 µg/mL) for 24 or 72 hours. Ex vivo porcine coronary artery rings were used to measure endothelial cell–dependent vascular smooth muscle cell relaxation after CPP exposure. Serum samples from an early chronic kidney disease cohort (n=245) were analyzed for calcification propensity (measure for CPP formation) and nitrate and nitrite levels (NOx).

RESULTS:
CPP exposure for 24 hours reduced eNOS (endothelial nitric oxide synthase) mRNA expression and decreased nitrite production, indicating reduced nitric oxide bioavailability. Also, 24-hour CPP exposure caused increased mitochondria-derived superoxide generation, together with nitrotyrosine protein residue formation. Long-term (72 hours) exposure of human umbilical vein endothelial cells to CPPs induced eNOS uncoupling and decreased eNOS protein expression, indicating further impairment of the nitric oxide pathway. The ex vivo porcine coronary artery model showed a significant reduction in endothelial-dependent vascular smooth muscle cell relaxation after CPP exposure. A negative association was observed between NOx levels and calcification propensity (r=−0.136; P=0.049) in sera of (early) chronic kidney disease patients.

CONCLUSIONS:
CPPs cause endothelial cell dysfunction by impairing nitric oxide metabolism and generating oxidative stress. Our findings provide new evidence for direct effects of CPPs on ECs and pathways involved.
Original languageEnglish
Pages (from-to)443–455
Number of pages13
JournalArteriosclerosis, thrombosis, and vascular biology
Volume43
Issue number3
DOIs
Publication statusPublished - 2-Feb-2023

Keywords

  • Humans
  • Animals
  • Swine
  • Nitric Oxide/metabolism
  • Nitrites/metabolism
  • Endothelium/metabolism
  • Nitric Oxide Synthase Type III/metabolism
  • Vascular Diseases
  • Human Umbilical Vein Endothelial Cells/metabolism
  • Renal Insufficiency, Chronic/metabolism
  • Endothelium, Vascular/metabolism

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