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Zinc Inhibits Apoptosis and Maintains NEP Downregulation, Induced by Ropivacaine, in HaCaT Cells

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Abstract

Zinc (Zn), a cell-protective metal against various toxic compounds, is the key agent for neutral endopeptidase (NEP) functional structure. NEP is a zinc metalloenzyme which degrades endogenous opioids and is expressed in human keratinocytes (HaCaT). Ropivacaine, a widely used opiate local anaesthetic, exerts cell toxic and apoptotic effects against HaCaT cells. The aim of the present study is to investigate whether zinc modulates the effects of ropivacaine on proliferation, viability, apoptosis and NEP expression in HaCaT cells. To investigate the role of ropivacaine in NEP function, HaCaT cells overexpressing NEP were generated via cell transfection with plasmids carrying NEP cDNA. Ropivacaine's anti-proliferative effect was tested by Neubauer's chamber cell counting, and induction of cell death was demonstrated by trypan blue exclusion assay. Apoptosis due to ropivacaine was tested via DNA fragmentation and poly-ADP-ribose-polymerase (PARP) cleavage. NEP and PARP expression was performed by western blot analysis. Results showed that zinc (15 μΜ) inhibited proliferation and cell death induction by ropivacaine (0.5, 1 and 2 mM) (p < 0.05) as well as apoptosis induced by the drug (0.5 and 1 mM) in HaCaT cells. Ropivacaine (1.0, 2.0 and 5.0 mM) downregulated NEP expression in the presence of zinc (15 μΜ) while NEP overexpression enhanced ropivacaine's apoptotic effect. In conclusion, the abilities of zinc to inhibit the toxic and apoptotic effects of ropivacaine, to maintain NEP downregulation induced by the drug and, consequently, to enhance its anaesthetic result suggest that zinc may have a significant role in pain management and tissue protection.

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Acknowledgments

This work was partially supported by a European's Society for Regional Anaesthesia (ESRA) Research Grant.

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Authors and Affiliations

  1. Laboratory of Physiology, Medical School, University of Ioannina, 45110, Ioannina, Greece

    Evangelos Kontargiris & Vasiliki Kalfakakou

  2. Department of Anaesthesia, Pain Relief and Palliative Care, University Clinic, Areteion Hospital, Athens, Greece

    Athina Vadalouka

  3. Department of Maxillofacial Surgery, Medical School, University of Ioannina, 45110, Ioannina, Greece

    Vasilios Ragos

Authors
  1. Evangelos Kontargiris
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  2. Athina Vadalouka
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  3. Vasilios Ragos
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  4. Vasiliki Kalfakakou
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Correspondence to Vasiliki Kalfakakou.

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Kontargiris, E., Vadalouka, A., Ragos, V. et al. Zinc Inhibits Apoptosis and Maintains NEP Downregulation, Induced by Ropivacaine, in HaCaT Cells. Biol Trace Elem Res 150, 460–466 (2012). https://doi.org/10.1007/s12011-012-9492-8

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  • DOI: https://doi.org/10.1007/s12011-012-9492-8

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