PMID- 17717122
OWN - NLM
STAT- MEDLINE
DCOM- 20080108
LR  - 20130926
IS  - 8750-7587 (Print)
IS  - 0161-7567 (Linking)
VI  - 103
IP  - 5
DP  - 2007 Nov
TI  - Vascular endothelial sampling and analysis of gene transcripts: a new 
      quantitative approach to monitor vascular inflammation.
PG  - 1873-8
AB  - BACKGROUND: Limited access to endothelial tissue is a major constraint when 
      investigating the cellular mechanisms of vascular inflammation in patients with 
      cardiovascular and metabolic diseases. We introduce venous endothelial sampling 
      coupled to quantitative analysis of gene transcripts by real-time PCR, as a novel 
      approach to study endothelial gene expression in human subjects. METHODS: 
      Endothelial cells were collected from a superficial forearm vein using five guide 
      wires sequentially inserted through a 20-gauge angiocatheter in seven patients 
      with history of cardiovascular events related to advanced vascular disease and in 
      17 healthy subjects. Endothelial cells were purified using magnetic beads coated 
      with endothelial specific antibodies. Endothelial mRNA was amplified using 
      RiboAmp HS RNA Amplification kit (Molecular Devices, Sunnyvale, CA). Amplified 
      RNA was analyzed by real-time PCR. RESULTS: Linearity of RNA amplification was 
      validated by real-time PCR using RNA from 1,000 human umbilical endothelial cells 
      (HUVECs) before and after amplification. In human subjects, vascular disease was 
      associated with significant induction of proatherosclerotic genes: early growth 
      response gene product (Egr-1) and monocyte chemoattractant protein-1 (MCP-1). 
      CONCLUSION: Venous endothelial sampling coupled to real-time PCR analysis is a 
      minimally invasive, safe, and reliable technique to monitor vascular inflammation 
      in human subjects. Expression of genes implicated in the atherosclerotic process 
      is increased in the venous endothelium of patients with arterial vascular 
      disease. Venous endothelial sampling and quantitative analysis of gene expression 
      may help develop new vascular-targeted biomarkers to identify and track the 
      impact of disease states and therapeutic interventions in vascular diseases.
FAU - Onat, Duygu
AU  - Onat D
AD  - Department of Medicine, Columbia Univ., New York, NY, USA.
FAU - Jelic, Sanja
AU  - Jelic S
FAU - Schmidt, Ann Marie
AU  - Schmidt AM
FAU - Pile-Spellman, John
AU  - Pile-Spellman J
FAU - Homma, Shunichi
AU  - Homma S
FAU - Padeletti, Margherita
AU  - Padeletti M
FAU - Jin, Zhezhen
AU  - Jin Z
FAU - Le Jemtel, Thierry H
AU  - Le Jemtel TH
FAU - Colombo, Paolo C
AU  - Colombo PC
FAU - Feng, Lei
AU  - Feng L
LA  - eng
GR  - K23-HL-72758/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20070823
PL  - United States
TA  - J Appl Physiol (1985)
JT  - Journal of applied physiology (Bethesda, Md. : 1985)
JID - 8502536
RN  - 0 (CCL2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Early Growth Response Protein 1)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Aged
MH  - Cardiovascular Diseases/*genetics/metabolism
MH  - Case-Control Studies
MH  - *Cell Separation
MH  - Cells, Cultured
MH  - Chemokine CCL2/analysis/genetics
MH  - Early Growth Response Protein 1/analysis/genetics
MH  - Endothelium, Vascular/*chemistry
MH  - Female
MH  - Gene Expression Profiling/*methods
MH  - Humans
MH  - Inflammation/*genetics/metabolism
MH  - Male
MH  - Middle Aged
MH  - *Polymerase Chain Reaction
MH  - RNA, Messenger/*analysis
MH  - Reproducibility of Results
MH  - *Transcription, Genetic
EDAT- 2007/08/25 09:00
MHDA- 2008/01/09 09:00
CRDT- 2007/08/25 09:00
PHST- 2007/08/25 09:00 [pubmed]
PHST- 2008/01/09 09:00 [medline]
PHST- 2007/08/25 09:00 [entrez]
AID - 00367.2007 [pii]
AID - 10.1152/japplphysiol.00367.2007 [doi]
PST - ppublish
SO  - J Appl Physiol (1985). 2007 Nov;103(5):1873-8. doi: 
      10.1152/japplphysiol.00367.2007. Epub 2007 Aug 23.