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Cyclic AMP and Cyclic GMP suppress TNFα-induced hepatocyte apoptosis by inhibiting FADD up-regulation via a protein kinase A-dependent pathway

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Abstract

Cyclic AMP (cAMP) and cyclic GMP (cGMP) suppress apoptosis in many cell types, including hepatocytes. We have previously shown that membrane-permeable cAMP and cGMP analogs attenuate tumor necrosis factor α plus actinomycin D (TNFα/ActD)-induced apoptosis in hepatocytes at a step upstream of caspase activation and cytochrome c release. Recently we have also shown that FADD levels increase 10 folds in response to TNFα/ActD. Therefore we hypothesized that cAMP and cGMP would inhibit FADD upregulation. We show here that cyclic nucleotide analogs dibutyryl cAMP (db-cAMP) and 8-bromo-cGMP (Br-cGMP) inhibit cell death and the cleavages of multiple caspases including caspase-10, -9, -8, -3, and -2, as well as suppress FADD protein up-regulation in TNFα/ActD-induced apoptosis. The inhibitory effects of cAMP were seen at lower concentrations than cGMP. Both cAMP and cGMP prevented FADD overexpression and cell death in hepatocytes transfected with the FADD gene. A protein kinase A (PKA) inhibitor, KT 5720, reversed the inhibition of FADD protein levels induced by cAMP or cGMP. In conclusion, our findings indicate that cAMP and cGMP prevent TNFα/ActD-induced apoptosis in hepatocytes and that this occurs in association with a near complete inhibition of the upregulation of FADD via a PKA-dependent mechanism.

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Abbreviations

TNFα:

tumor necrosis factor-α

ActD:

actinomycin D

FADD:

Fas-associated death domain protein

db-cAMP:

dibutyryl cyclic adenosine monophosphate

Br-cGMP:

8-bromoguanosine 3′,5′-cyclic monophosphate

PKA.:

protein kinase A; Z-VAD-fmk, benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone

Ac-IETD-CHO:

Ac-Ile-Glu-Thr-Asp-CHO

(Sρ)-cAMPS:

Adenosine 3′,5′-cyclic monophosphorothioate, Sp-isomer

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP-nick end labeling

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Authors and Affiliations

  1. Department of Surgery, School of Medicine, University of Pittsburgh, 3459 Fifth Ave., NW607, MUH, Pittsburgh, PA, 15213, USA

    Y. Wang, Peter K. M. Kim, X. Peng, P. Loughran, Y. Vodovotz, B. Zhang & T. R. Billiar

  2. Department of Surgery, School of Medicine, University of Pittsburgh, P.O.Box 7533, Pittsburgh, PA, 15213, USA

    T. R. Billiar

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  1. Y. Wang
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  2. Peter K. M. Kim
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  3. X. Peng
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Correspondence to T. R. Billiar.

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Supported by the National Institutes of Health Grant GM-44100 (to T.R.B).

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Wang, Y., Kim, P.K.M., Peng, X. et al. Cyclic AMP and Cyclic GMP suppress TNFα-induced hepatocyte apoptosis by inhibiting FADD up-regulation via a protein kinase A-dependent pathway. Apoptosis 11, 441–451 (2006). https://doi.org/10.1007/s10495-005-4293-6

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  • DOI: https://doi.org/10.1007/s10495-005-4293-6

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