In vitro effects of waterpipe smoke condensate on endothelial cell function: a potential risk factor for vascular disease

Toxicol Lett. 2013 May 23;219(2):133-42. doi: 10.1016/j.toxlet.2013.02.015. Epub 2013 Feb 28.


Aim: Despite its increasing popularity, little is known about the health effects of waterpipe smoking (WPS), particularly on the cardiovascular system. To investigate the role of WPS as a risk factor for vascular disease, we evaluated its effect on endothelial cell function, which is an early event in vascular disease pathogenesis. We assessed the changes in cell viability, ROS generation, inflammatory and vasodilatory markers and in vitro angiogenesis of human aortic endothelial cells in response to waterpipe smoke condensate exposure.

Methods and results: Mainstream waterpipe smoke condensate (WSC) was generated using a standard laboratory machine protocol. Compared to control, WSC induced cell cycle arrest, apoptosis, and oxidative stress in human primary endothelial cells. In addition, we assayed for impaired endothelium-dependent vasodilation and induced inflammation by studying the effect of WPS on the content and activity of AMPK, eNOS proteins and NF-κB p65 ser536 phosphorylation, respectively. WSC inhibited AMPK/eNOS phosphorylation and induced phosphorylation of p65. Moreover, we evaluated endothelial cells repair mechanism related properties that include migration/invasion and in vitro tube formation upon treatment with WSC. WSC reduced the motility and inhibited angiogenic potential of HAEC cells.

Conclusions: WPS induced endothelial cell dysfunction as evident by exerting oxidative stress, inflammation, and impaired endothelial vasodilatory function and repair mechanisms. All together these data provide evidence for the potential contribution of WPS to endothelial dysfunction and thus to vascular disease.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • AMP-Activated Protein Kinases / metabolism
  • Annexin A5 / metabolism
  • Blotting, Western
  • Cell Cycle / drug effects
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Endothelial Cells / drug effects*
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects*
  • Flow Cytometry
  • Humans
  • Inflammation / pathology
  • Neovascularization, Physiologic / drug effects
  • Nitric Oxide Synthase Type III / metabolism
  • Reactive Oxygen Species / metabolism
  • Risk Factors
  • Smoke / adverse effects*
  • Smoke / analysis*
  • Smoking*
  • Tobacco / chemistry*
  • Vascular Diseases / chemically induced*
  • Water


  • Annexin A5
  • Reactive Oxygen Species
  • Smoke
  • Water
  • Nitric Oxide Synthase Type III
  • AMP-Activated Protein Kinases