PMID- 24462831 OWN - NLM STAT- MEDLINE DCOM- 20141020 LR - 20240410 IS - 1095-8584 (Electronic) IS - 0022-2828 (Print) IS - 0022-2828 (Linking) VI - 68 DP - 2014 Mar TI - BDNF secretion by human pulmonary artery endothelial cells in response to hypoxia. PG - 89-97 LID - 10.1016/j.yjmcc.2014.01.006 [doi] AB - Within human pulmonary artery, neurotrophin growth factors [NTs; e.g. brain-derived neurotrophic factor (BDNF)] and their high-affinity receptors (tropomyosin-related kinase; Trk) and low-affinity receptors p75 neurotrophin receptor (p75NTR) have been reported, but their functional role is incompletely understood. We tested the hypothesis that BDNF is produced by human pulmonary artery endothelial cells (PAECs). In the context of hypoxia as a risk factor for pulmonary hypertension, we examined the effect of hypoxia on BDNF secretion and consequent autocrine effects on pulmonary endothelium. Initial ELISA analysis of circulating BDNF in 30 healthy human volunteers showed that 72 h exposure to high altitude (~11,000 ft, alveolar PO2 = 100 mmHg) results in higher BDNF compared to samples taken at sea level. Separately, in human PAECs exposed for 24h to normoxia vs. hypoxia (1-3% O2), ELISA of extracellular media showed increased BDNF levels. Furthermore, quantitative PCR of PAECs showed 3-fold enhancement of BDNF gene transcription with hypoxia. In PAECs, BDNF induced NO production (measured using an NO-sensitive fluorescent dye DAF2-DA) that was significantly higher under hypoxic conditions, an effect also noted with the TrkB agonist 7,8-DHF. Importantly, hypoxia-induced NO was blunted by neutralization of secreted BDNF using the chimeric TrkB-Fc. Both hypoxia and BDNF increased iNOS (but not eNOS) mRNA expression. In accordance, BDNF enhancement of NO in hypoxia was not blunted by 50 nM L-NAME (eNOS inhibition) but substantially lower with 100 muM L-NAME (eNOS and iNOS inhibition). Hypoxia and BDNF also induced expression of hypoxia inducible factor 1 alpha (HIF-1alpha), a subunit of the transcription factor HIF-1, and pharmacological inhibition of HIF-1 diminished hypoxia effects on BDNF expression and secretion, and NO production. These results indicate that human PAECs express and secrete BDNF in response to hypoxia via a HIF-1-regulated pathway. CI - Copyright (c) 2014 Elsevier Ltd. All rights reserved. FAU - Helan, Martin AU - Helan M AD - Department of Anesthesiology, Mayo Clinic, Rochester. AD - ICRC, International Clinical Research Center, Brno, Czech Republic. AD - Department of Anesthesiology and Intensive Care, St. Anne's University Hospital and Faculty of Medicine, Masaryk University, Brno, Czech Republic. FAU - Aravamudan, Bharathi AU - Aravamudan B AD - Department of Anesthesiology, Mayo Clinic, Rochester. FAU - Hartman, William R AU - Hartman WR AD - Department of Anesthesiology, Mayo Clinic, Rochester. AD - Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester. FAU - Thompson, Michael A AU - Thompson MA AD - Department of Anesthesiology, Mayo Clinic, Rochester. FAU - Johnson, Bruce D AU - Johnson BD AD - Cardiovascular diseases, Mayo Clinic, Rochester. FAU - Pabelick, Christina M AU - Pabelick CM AD - Department of Anesthesiology, Mayo Clinic, Rochester. AD - Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester. FAU - Prakash, Y S AU - Prakash YS AD - Department of Anesthesiology, Mayo Clinic, Rochester. AD - Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester. LA - eng GR - HL056470/HL/NHLBI NIH HHS/United States GR - R01 HL056470/HL/NHLBI NIH HHS/United States GR - R01 HL088029/HL/NHLBI NIH HHS/United States GR - UL1 RR024150/RR/NCRR NIH HHS/United States GR - NCRR 1UL1 RR024150/PHS HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20140123 PL - England TA - J Mol Cell Cardiol JT - Journal of molecular and cellular cardiology JID - 0262322 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Hypoxia-Inducible Factor 1) RN - 0 (Membrane Glycoproteins) RN - 0 (RNA, Messenger) RN - 31C4KY9ESH (Nitric Oxide) RN - 7171WSG8A2 (BDNF protein, human) RN - EC 1.14.13.39 (NOS2 protein, human) RN - EC 1.14.13.39 (NOS3 protein, human) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type III) RN - EC 2.7.- (Protein Kinases) RN - EC 2.7.10.1 (Protein-Tyrosine Kinases) RN - EC 2.7.10.1 (Receptor, trkB) RN - EC 2.7.10.1 (tropomyosin-related kinase-B, human) RN - EC 3.5.3.1 (ARG1 protein, human) RN - EC 3.5.3.1 (ARG2 protein, human) RN - EC 3.5.3.1 (Arginase) SB - IM MH - Arginase/metabolism MH - Brain-Derived Neurotrophic Factor/blood/genetics/*metabolism MH - Cell Hypoxia MH - Cells, Cultured MH - Endothelial Cells/*metabolism MH - Endothelium, Vascular/pathology MH - Gene Expression MH - Humans MH - Hypoxia/blood MH - Hypoxia-Inducible Factor 1/metabolism MH - Membrane Glycoproteins MH - Nitric Oxide/metabolism MH - Nitric Oxide Synthase Type II/metabolism MH - Nitric Oxide Synthase Type III/metabolism MH - Protein Kinases/metabolism MH - Protein-Tyrosine Kinases MH - Pulmonary Artery/pathology MH - RNA, Messenger/genetics/metabolism MH - Receptor, trkB MH - Signal Transduction PMC - PMC3977651 MID - NIHMS559971 OTO - NOTNLM OT - Hypoxia inducible factor 1 OT - Neurotrophin OT - Nitric oxide OT - Tropomyosin related kinase OT - eNOS OT - iNOS EDAT- 2014/01/28 06:00 MHDA- 2014/10/21 06:00 PMCR- 2015/03/01 CRDT- 2014/01/28 06:00 PHST- 2013/09/03 00:00 [received] PHST- 2013/12/24 00:00 [revised] PHST- 2014/01/14 00:00 [accepted] PHST- 2014/01/28 06:00 [entrez] PHST- 2014/01/28 06:00 [pubmed] PHST- 2014/10/21 06:00 [medline] PHST- 2015/03/01 00:00 [pmc-release] AID - 10.1016/j.yjmcc.2014.01.006 [doi] PST - ppublish SO - J Mol Cell Cardiol. 2014 Mar;68:89-97. doi: 10.1016/j.yjmcc.2014.01.006. Epub 2014 Jan 23.