title: Identification of a novel pathway of transforming growth factor-β1 regulation by extracellular NAD+ in mouse macrophages: in vitro and in silico studies.
abstract: Extracellular β-nicotinamide adenine dinucleotide (NAD(+)) is anti-inflammatory. We hypothesized that NAD(+) would modulate the anti-inflammatory cytokine Transforming Growth Factor (TGF)-β1. Indeed, NAD(+) led to increases in both active and latent cell-associated TGF-β1 in RAW 264.7 mouse macrophages as well as in primary peritoneal macrophages isolated from both C3H/HeJ (TLR4-mutant) and C3H/HeOuJ (wild-type controls for C3H/HeJ) mice. NAD(+) acts partially via cyclic ADP-ribose (cADPR) and subsequent release of Ca(2+). Treatment of macrophages with the cADPR analog 3-deaza-cADPR or Ca(2+) ionophores recapitulated the effects of NAD(+) on TGF-β1, whereas the cADPR antagonist 8-Br-cADPR, Ca(2+) chelation, and antagonism of L-type Ca(2+) channels suppressed these effects. The time and dose effects of NAD(+) on TGF-β1 were complex and could be modeled both statistically and mathematically. Model-predicted levels of TGF-β1 protein and mRNA were largely confirmed experimentally but also suggested the presence of other mechanisms of regulation of TGF-β1 by NAD(+). Thus, in vitro and in silico evidence points to NAD(+) as a novel modulator of TGF-β1.
authors: Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, USA. zamorar@pitt.edu
authors: Ruben
authors: R
authors: Zamora
authors: Nabil
authors: N
authors: Azhar
authors: Rajaie
authors: R
authors: Namas
authors: Mallikarjuna R
authors: MR
authors: Metukuri
authors: Thierry
authors: T
authors: Clermont
authors: Chase
authors: C
authors: Gladstone
authors: Rami A
authors: RA
authors: Namas
authors: Linda
authors: L
authors: Hermus
authors: Cristina
authors: C
authors: Megas
authors: Gregory
authors: G
authors: Constantine
authors: Timothy R
authors: TR
authors: Billiar
authors: Mitchell P
authors: MP
authors: Fink
authors: Yoram
authors: Y
authors: Vodovotz
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D061207:Calcium Ionophores / Q000494:pharmacology; D002460:Cell Line; D036563:Cyclic ADP-Ribose / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008817:Mice, Mutant Strains; D008954:Models, Biological; D009243:NAD / Q000235:genetics / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D061207:Calcium Ionophores; C508961:Tgfb1 protein, mouse; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D053773:Transforming Growth Factor beta1; D009243:NAD; D036563:Cyclic ADP-Ribose; D002118:Calcium