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Coupling endoplasmic reticulum stress to cell death program in isolated human pancreatic islets: effects of gene transfer of Bcl-2

  • Original Article
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Transplant International

Abstract

A variety of toxic insults can result in endoplasmic reticulum (ER)-stress that ultimately leads to apoptosis. β-cells have a highly developed ER due to a great commitment to insulin production. The present study was carried out to determine the role of ER-stress in isolated human pancreatic islet apoptosis, and the potential protective effects of Bcl-2. Isolated human islets were infected with an adenoviral vector encoding Bcl-2 and then exposed to brefeldin-A, tunicamycin, A23187 and pro-inflammatory cytokines. Activation of caspase-12 was analyzed by means of Western blots. Apoptosis was evaluated using a commercial quantitative assay. ER-stress-inducers promoted caspase-12 activation and apoptosis, effect reversed by overexpression of Bcl-2. Co-localization of caspase-12 and Bcl-2 in the microsomal islet fractions were demonstrated by means of Western blots. We can conclude that the current studies highlight the importance of Bcl-2 as an anti-apoptotic protein, and shed new light on the mechanisms underlying its cytoprotective effects on pancreatic islets.

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Acknowledgments

We thank Stacie M. Jenkins, BS, and Barry Grace, BS, CPTC, for their technical assistance during the human islet isolations. This paper was supported by research grants from Diabetes Trust Fund and Protective Life Clinical Initiative Award, University of Alabama at Birmingham.

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Authors and Affiliations

  1. Division of Transplantation, University of Alabama at Birmingham, 748 Lyons-Harrison Research Building, 701 19th Street South, Birmingham, AL 35294, USA

    Juan L. Contreras, Cheryl A. Smyth, Christopher Eckstein, Carlton J. Young, J. Anthony Thompson & Devin E. Eckhoff

  2. Division of Human Gene Therapy, University of Alabama at Birmingham, Birmingham, Alabama, USA

    Guadalupe Bilbao & David T. Curiel

Authors
  1. Juan L. Contreras
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  2. Cheryl A. Smyth
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  3. Guadalupe Bilbao
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  4. Christopher Eckstein
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  5. Carlton J. Young
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  6. J. Anthony Thompson
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  7. David T. Curiel
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  8. Devin E. Eckhoff
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Correspondence to Juan L. Contreras.

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Contreras, J.L., Smyth, C.A., Bilbao, G. et al. Coupling endoplasmic reticulum stress to cell death program in isolated human pancreatic islets: effects of gene transfer of Bcl-2. Transpl Int 16, 537–542 (2003). https://doi.org/10.1007/s00147-003-0619-x

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  • DOI: https://doi.org/10.1007/s00147-003-0619-x

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