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27 February 2018 : Original article  

Treatment of Donor Rat Hearts Prior to Transplantation with FLIP (FADD-Like Interleukin Beta-Converting Enzyme (FLICE)-Like Inhibitory Protein) in Cardioplegic Solution Decreased Apoptosis at Thirty Minutes Post-transplantation and Decreased Total Tyrosine Phosphorylation Levels

Feliu Roset1ABCDE, Jesus M. Ureña2AEF*, Tiziana Cotrufo2BCDE, José Carreras1ACD, Pablo Pérez de la Ossa13ABCDEFG, Fernando Climent1ACDEFG

DOI: 10.12659/AOT.903913

Ann Transplant 2018; 23:144-152


BACKGROUND: Heart transplantation is a therapeutic option for patients with severe coronary artery disease or heart failure. One of the difficulties to overcome is the apoptosis of cardiomyocytes in the donor organ. To prevent apoptosis in the donor organ, we developed a fusion protein containing FLIP (FADD-like interleukin beta-converting enzyme (FLICE)-like inhibitory protein) to inhibit caspase-8.

MATERIAL AND METHODS: We linked the cDNA coding for the FLIP protein to the transduction domain of HIV (human immunodeficiency virus) to allow the protein to enter cells. The recombinant protein was used at two different concentrations, 3 nM and 30 nM, for treatment of the donor heart in rat transplantation experiments. After transplantation, apoptosis was measured by ELISA, and the levels of active caspase-3, caspase-8, Bid, and PUMA were determined by western blotting using specific antibodies.

RESULTS: We observed that treatment of the donor organ with a solution containing this protein reduced the apoptosis level in the donor organ after 30 minutes post-transplantation as measured by the total of apoptotic cells with ELISA assay, and caspase-8 and caspase-3 activation and decreased levels of BH3-only proteins such as Bid and PUMA. Furthermore, this treatment also reduced the total tyrosine phosphorylation levels, which may be a possible measurement of lower oxidative stress levels in cardiomyocytes.

CONCLUSIONS: Protein FLIP solution reduced apoptosis at 30 minutes post-transplantation and decreased levels of several regulators of apoptosis.

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Annals of Transplantation eISSN: 2329-0358
Annals of Transplantation eISSN: 2329-0358