Cyclosporine A–protection against microvascular hyperpermeability is calcineurin independent

Childs, Ed W.; Tharakan, Binu; Nurudeen, Suliat; Delmas, Thomas L.; Hellman, Joseph; Christie, Tasheika; Hunter, Felicia A.; Smythe, W. Roy

doi:10.1016/j.amjsurg.2009.11.002

« Previous Next »The American Journal of Surgery
Volume 199, Issue 4 , Pages 542-548, April 2010

Cyclosporine A–protection against microvascular hyperpermeability is calcineurin independent

Ed W. Childs, M.D.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA
- Corresponding author. Tel.: +1-254-724-5638; fax: +1-254-724-7912
Binu Tharakan, Ph.D.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA
Suliat Nurudeen, B.S.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA
Thomas L. Delmas, B.S.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA
Joseph Hellman, B.S.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA
Tasheika Christie, B.S.
- Undergraduate Medical Academy, Prairie View A&M University, Prairie View, TX, USA
Felicia A. Hunter, B.S.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA
W. Roy Smythe, M.D.
- Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA

Received 1 July 2009; received in revised form 20 November 2009

Abstract

Background

Mitochondria-mediated apoptotic signaling contributes to microvascular hyperpermeability. We hypothesized that cyclosporine A (CsA), which protects mitochondrial transition pores, would attenuate hyperpermeability independent of its calcineurin inhibitory property.

Methods

Hyperpermeability was induced in microvascular endothelial cell monolayers using proapoptotic BAK or active caspase-3 after CsA or a specific calcineurin inhibitor, calcineurin autoinhibitory peptide (CIP), treatment. Permeability was measured based on fluorescein isothiocyanate–albumin flux across the monolayers. Mitochondrial transmembrane potential (MTP) was determined using 5,5′,6,6′-tetrachoro-1,1′,3,3′-tetraethylbenzimidazolyl carbocyanine iodide. Mitochondrial release of cytochrome c was measured using an enzyme-linked immunosorbent assay and caspase-3 activity fluorometrically.

Results

CsA-attenuated (10 nmol/L) but not CIP-attenuated (100 μmol/L) BAK induced hyperpermeability (P < .05), CsA- but not CIP-attenuated BAK induced a decrease in MTP and an increase in cytochrome c levels and caspase-3 activity (P < .05). CsA and CIP were ineffective against caspase-3–induced hyperpermeability.

Conclusions

CsA attenuated hyperpermeability by protecting MTP, thus preventing mitochondria-mediated apoptotic signaling. The protective effect of CsA is independent of calcineurin inhibition.

Keywords: Vascular permeability, Cell death, “Intrinsic” apoptotic cascade