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Detergent sclerosants at sub-lytic concentrations induce endothelial cell apoptosis through a caspase dependent pathway

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Abstract

To investigate the apoptotic effects of detergent sclerosants sodium tetradecylsulphate (STS) and polidocanol (POL) on endothelial cells at sub-lytic concentrations. Human umbilical vein endothelial cells (HUVECs) were isolated and labelled with antibodies to assess for apoptosis and examined with confocal microscopy and flow cytometry. Isolated HUVECs viability was assessed using propidium iodide staining. Early apoptosis was determined by increased phosphatidylserine exposure by lactadherin binding. Caspase 3, 8, 9 and Bax activation as well as inhibitory assays with Pan Caspase (Z-VAD-FMK) and Bax (BI-6C9) were assessed to identify apoptotic pathways. Porimin activation was used to assess cell membrane permeability. Cell lysis reached almost 100 % with STS at 0.3 % and with POL at 0.6 %. Apoptosis was seen with both STS and POL at concentrations ranging from 0.075 to 0.15 %. PS exposure increased with both STS and POL and exhibited a dose-dependent trend. Active Caspase 3, 8 and 9 but not Bax were increased in HUVECs stimulated with low concentrations of both STS and POL. Inhibitory assays demonstrated Caspase 3, 8, 9 inhibition at low concentrations (0.075 to 0.6 %) with both STS and POL. Both agents increased the activation of porimin at all concentrations. Both sclerosants induced endothelial cell (EC) apoptosis at sub-lytic concentrations through a caspase-dependant pathway. Both agents induced EC oncosis.

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Acknowledgments

This research was funded by the Academy of Phlebology and supported by the Dermatology, Phlebology and Fluid Mechanics Research Laboratory, St Vincent’s Centre for Applied Medical Research, Sydney, Australia.

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Authors and Affiliations

  1. Dermatology, Phlebology and Fluid Mechanics Research Laboratory, St Vincent’s Centre for Applied Medical Research, Level 8, Lowy-Packer Building, 405 Liverpool Street, Darlinghurst, Sydney, NSW, 2010, Australia

    Osvaldo Cooley-Andrade, David Ewan Connor & Kurosh Parsi

  2. The University of New South Wales, Sydney, Australia

    Osvaldo Cooley-Andrade, Kelvin Cheung, An-Ning Chew, David Ewan Connor & Kurosh Parsi

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  1. Osvaldo Cooley-Andrade
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  2. Kelvin Cheung
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  3. An-Ning Chew
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  5. Kurosh Parsi
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Correspondence to Kurosh Parsi.

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Cooley-Andrade, O., Cheung, K., Chew, AN. et al. Detergent sclerosants at sub-lytic concentrations induce endothelial cell apoptosis through a caspase dependent pathway. Apoptosis 21, 836–845 (2016). https://doi.org/10.1007/s10495-016-1252-3

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