PMID- 19193655
OWN - NLM
STAT- MEDLINE
DCOM- 20090817
LR  - 20211203
IS  - 1755-3245 (Electronic)
IS  - 0008-6363 (Linking)
VI  - 82
IP  - 3
DP  - 2009 Jun 1
TI  - TGF-beta1 inhibits expression and activity of hENT1 in a nitric oxide-dependent 
      manner in human umbilical vein endothelium.
PG  - 458-67
LID - 10.1093/cvr/cvp045 [doi]
AB  - AIMS: We studied whether transforming growth factor beta1 (TGF-beta1) modulates 
      human equilibrative nucleoside transporters 1 (hENT1) expression and activity in 
      human umbilical vein endothelial cells (HUVECs). hENT1-mediated adenosine 
      transport and expression are reduced in gestational diabetes and hyperglycaemia, 
      conditions associated with increased synthesis and release of nitric oxide (NO) 
      and TGF-beta1 in this cell type. TGF-beta1 increases NO synthesis via activation 
      of TGF-beta receptor type II (TbetaRII), and NO inhibits hENT1 expression and 
      activity in HUVECs. METHODS AND RESULTS: HUVECs (passage 2) were used for 
      experiments. Total and hENT1-mediated adenosine transport was measured in the 
      absence or presence of TGF-beta1, NG-nitro-L-arginine methyl ester (L-NAME, NO 
      synthase inhibitor), S-nitroso-N-acetyl-L,D-penicillamine (SNAP, NO donor), 
      and/or KT-5823 (protein kinase G inhibitor) in control cells and cells expressing 
      a truncated form of TGF-beta1 receptor type II (TTbetaRII). Western blot and 
      real-time PCR were used to determine hENT1 protein abundance and mRNA expression. 
      SLC29A1 gene promoter and specific protein 1 (Sp1) transcription factor activity 
      was assayed. Vascular reactivity was assayed in endothelium-intact or -denuded 
      umbilical vein rings. TGF-beta1 reduced hENT1-mediated adenosine transport, hENT1 
      protein abundance, hENT1 mRNA expression, and SLC29A1 gene promoter activity, but 
      increased Sp1 binding to DNA. TGF-beta1 effect was blocked by L-NAME and KT-5823 
      and mimicked by SNAP in control cells. However, TGF-beta1 was ineffective in 
      cells expressing TTbetaRII or a mutated Sp1 consensus sequence. Vasodilatation in 
      response to TGF-beta1 and S-(4-nitrobenzyl)-6-thio-inosine (an ENT inhibitor) was 
      endothelium-dependent and blocked by KT-5823 and ZM-241385. CONCLUSION: hENT1 is 
      down-regulated by activation of TbetaRII by TGF-beta1 in HUVECs, a phenomenon 
      where NO and Sp1 play key roles. These findings comprise physiological mechanisms 
      that could be important in diseases where TGF-beta1 plasma level is increased as 
      in gestational diabetic mothers or patients with diabetes mellitus.
FAU - Vega, Jose L
AU  - Vega JL
AD  - Cellular and Molecular Physiology Laboratory, Department of Obstetrics and 
      Gynaecology, Medical Research Centre (CIM), School of Medicine, Faculty of 
      Medicine, Pontificia Universidad Catolica de Chile, PO Box 114-D, Santiago, 
      Chile.
FAU - Puebla, Carlos
AU  - Puebla C
FAU - Vasquez, Rodrigo
AU  - Vasquez R
FAU - Farias, Marcelo
AU  - Farias M
FAU - Alarcon, Julio
AU  - Alarcon J
FAU - Pastor-Anglada, Marcal
AU  - Pastor-Anglada M
FAU - Krause, Bernardo
AU  - Krause B
FAU - Casanello, Paola
AU  - Casanello P
FAU - Sobrevia, Luis
AU  - Sobrevia L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090204
PL  - England
TA  - Cardiovasc Res
JT  - Cardiovascular research
JID - 0077427
RN  - 0 (Equilibrative Nucleoside Transporter 1)
RN  - 0 (Receptors, Transforming Growth Factor beta)
RN  - 0 (SLC29A1 protein, human)
RN  - 0 (TGFB1 protein, human)
RN  - 0 (Transforming Growth Factor beta1)
RN  - 31C4KY9ESH (Nitric Oxide)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.30 (Receptor, Transforming Growth Factor-beta Type II)
RN  - K72T3FS567 (Adenosine)
SB  - IM
MH  - Adenosine/*metabolism
MH  - Cells, Cultured
MH  - Endothelial Cells/metabolism
MH  - Endothelium, Vascular/*metabolism
MH  - Equilibrative Nucleoside Transporter 1/genetics/*metabolism
MH  - Female
MH  - Humans
MH  - Nitric Oxide/*metabolism
MH  - Pregnancy
MH  - Pregnancy Complications/metabolism
MH  - Promoter Regions, Genetic
MH  - Protein Serine-Threonine Kinases/*metabolism
MH  - Receptor, Transforming Growth Factor-beta Type II
MH  - Receptors, Transforming Growth Factor beta/*metabolism
MH  - Transcription, Genetic
MH  - Transforming Growth Factor beta1/*metabolism
MH  - Vasodilation
EDAT- 2009/02/06 09:00
MHDA- 2009/08/18 09:00
CRDT- 2009/02/06 09:00
PHST- 2009/02/06 09:00 [entrez]
PHST- 2009/02/06 09:00 [pubmed]
PHST- 2009/08/18 09:00 [medline]
AID - cvp045 [pii]
AID - 10.1093/cvr/cvp045 [doi]
PST - ppublish
SO  - Cardiovasc Res. 2009 Jun 1;82(3):458-67. doi: 10.1093/cvr/cvp045. Epub 2009 Feb 
      4.