PMID- 8569176
OWN - NLM
STAT- MEDLINE
DCOM- 19960304
LR  - 20061115
IS  - 0023-6837 (Print)
IS  - 0023-6837 (Linking)
VI  - 74
IP  - 1
DP  - 1996 Jan
TI  - Measurement by quantitative reverse transcription-polymerase chain reaction of 
      the levels of tumor necrosis factor alpha mRNA in atherosclerotic arteries in 
      Watanabe heritable hyperlipidemic rabbits.
PG  - 136-45
AB  - Tumor necrosis factor-alpha (TNF-alpha), produced by activated monocytes and 
      other cells, has been proposed as a mediator of importance in atherosclerosis. In 
      this study, we use a newly developed technique, quantitative reverse 
      transcription-polymerase chain reaction (RT-PCR), to measure the mRNA levels of 
      TNF-alpha in arteries of Watanabe Heritable Hyperlipidemic (WHHL) rabbits in 
      relation to the progression of atherosclerosis. Co-amplification of known amounts 
      of TNF-alpha RNA and TNF-alpha internal control RNA indicated that the 
      quantitative RT-PCR method was quite reliable, with a < 5% difference between 
      TNF-alpha mRNA levels deduced from the standard curve and actually loaded 
      TNF-alpha mRNA. As another control, TNF-alpha mRNA levels in 
      lipopolysaccharide-induced (LPS-induced) and uninduced monocytes were measured, 
      and a 9.3-fold increase in the TNF-alpha mRNA levels was observed in LPS-induced 
      monocytes. TNF-alpha mRNA levels in the aortic arches of healthy New Zealand 
      White (NZW) rabbits was 0.0112 -/+ 0.0016 (SD) pg per ng tissue RNA. TNF-alpha 
      mRNA levels in the aortic arches and descending thoracic aortas of 6-month-old 
      WHHL rabbits were 0.0273 -/+ 0.0066 pg and 0.0176 -/+ 0.0013 pg per ng tissue 
      RNA, respectively. TNF-alpha mRNA levels in the aortic arches and descending 
      thoracic aortas of 18-month-old WHHL rabbits were much higher than in those of 
      6-month-old WHHL rabbits, with values of 0.2107 -/+ 0.0205 pg and 0.1043 -/+ 
      0.0196 pg per ng tissue RNA, respectively. These findings indicate that: a) 
      Quantitative RT-PCR can be used to measure levels of small abundance RNA in 
      normal and diseased tissues accurately; b) no significant increase in TNF-alpha 
      mRNA levels was observed in the aortic arches and descending thoracic aortas of 
      6-month-old WHHL rabbits compared with those of NZW rabbits, although they had 
      multiple raised intimal lesions; and c) a significantly elevated total tissue 
      level of TNF-alpha mRNA is demonstrable late in the course of atherosclerosis in 
      18-month-old WHHL rabbits (Bonferroni method). These findings suggest that 
      TNF-alpha might play an important role in the progression of advanced 
      atherosclerosis, although total tissue levels of TNF-alpha mRNA are not 
      unequivocally elevated earlier in the course of the disease.
FAU - Lei, X
AU  - Lei X
AD  - Department of Pathology, University of Texas Medical School, Houston 77030, USA.
FAU - Buja, L M
AU  - Buja LM
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Lab Invest
JT  - Laboratory investigation; a journal of technical methods and pathology
JID - 0376617
RN  - 0 (RNA, Messenger)
RN  - 0 (Tumor Necrosis Factor-alpha)
SB  - IM
MH  - Animals
MH  - Aorta, Thoracic/*chemistry/pathology
MH  - Arteriosclerosis/*genetics/pathology
MH  - Base Sequence
MH  - Female
MH  - Hyperlipidemias/*genetics/pathology
MH  - Male
MH  - Molecular Sequence Data
MH  - Monocytes/chemistry
MH  - Plasmids
MH  - *Polymerase Chain Reaction/methods/standards
MH  - RNA, Messenger/*analysis
MH  - Rabbits
MH  - Reference Standards
MH  - Transcription, Genetic
MH  - Tumor Necrosis Factor-alpha/*genetics/standards
EDAT- 1996/01/01 00:00
MHDA- 1996/01/01 00:01
CRDT- 1996/01/01 00:00
PHST- 1996/01/01 00:00 [pubmed]
PHST- 1996/01/01 00:01 [medline]
PHST- 1996/01/01 00:00 [entrez]
PST - ppublish
SO  - Lab Invest. 1996 Jan;74(1):136-45.