PMID- 15081318
OWN - NLM
STAT- MEDLINE
DCOM- 20041119
LR  - 20081121
IS  - 0022-2828 (Print)
IS  - 0022-2828 (Linking)
VI  - 36
IP  - 4
DP  - 2004 Apr
TI  - Regulation of monocyte chemoattractant protein-1 by the oxidized lipid, 
      13-hydroperoxyoctadecadienoic acid, in vascular smooth muscle cells via nuclear 
      factor-kappa B (NF-kappa B).
PG  - 585-95
AB  - The leukocyte- type 12/15-Lipoxygenase (12/15-LO) enzyme and its oxidized lipid 
      products play important roles in vascular smooth muscle cell (VSMC) growth, 
      migration, and matrix responses associated with hypertension, atherosclerosis, 
      and restenosis. However, much less is known about their inflammatory effects. In 
      this study, we showed that the 12/15-LO product of linoleic acid, 
      13-hydroperoxyocta decadienoic acid (13-HPODE) can transcriptionally upregulate 
      the expression of the chemokine monocyte chemoattractant protein-1 (MCP-1) in 
      VSMC. We also observed reduced activation of the transcription factor, NF-kappa B 
      and reduced expression of MCP-1/JE mRNA in VSMC from 12/15-LO knock-out mice 
      relative to WT. To confirm the role of NF-kappa B in 13-HPODE-induced MCP-1 
      expression and to selectively block the induction of such inflammatory genes in 
      VSMC, we designed novel molecular approaches to knockdown NF-kappa B with short 
      interfering RNAs (siRNAs). We designed siRNAs to human NF-kappa B p65 
      transcriptionally active subunit by using a rapid PCR-based approach that 
      generates sense and antisense siRNA separated by a hairpin loop downstream of the 
      U6 promoter. siRNA PCR products targeting seven different sites on p65 cDNA could 
      induce upto 92% reduction in HA-p65 protein levels. A six-fold decrease in 
      NF-kappa B-dependent luciferase activity was also seen. Transfection of human 
      VSMC with these siRNA PCR products resulted in 70% reduction in p65 protein 
      levels. We cloned the PCR products into a pCR3.1 vector and these p65 siRNA 
      expressing plasmids very effectively blocked 13-HPODE-induced expression of both 
      MCP-1 and TNF-alpha genes. These results show for the first time that 13-HPODE 
      can induce MCP-1 in the vasculature via activation of NF-kappa B.
FAU - Dwarakanath, Roopashree S
AU  - Dwarakanath RS
AD  - Departments of Diabetes, Beckman Research Institute of City of Hope, Duarte, CA 
      91010, USA.
FAU - Sahar, Saurabh
AU  - Sahar S
FAU - Reddy, Marpadga A
AU  - Reddy MA
FAU - Castanotto, Daniela
AU  - Castanotto D
FAU - Rossi, John J
AU  - Rossi JJ
FAU - Natarajan, Rama
AU  - Natarajan R
LA  - eng
GR  - P01 HL55798/HL/NHLBI NIH HHS/United States
GR  - R01 DK58191/DK/NIDDK NIH HHS/United States
GR  - R01 DK65073/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - J Mol Cell Cardiol
JT  - Journal of molecular and cellular cardiology
JID - 0262322
RN  - 0 (Chemokine CCL2)
RN  - 0 (Linoleic Acids)
RN  - 0 (Lipid Peroxides)
RN  - 0 (NF-kappa B)
RN  - 0 (Oligonucleotides)
RN  - 0 (Oligonucleotides, Antisense)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Small Interfering)
RN  - 23017-93-8 (13-hydroperoxy-9,11-octadecadienoic acid)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Cell Line
MH  - Cells, Cultured
MH  - Chemokine CCL2/*biosynthesis/metabolism
MH  - Cloning, Molecular
MH  - Down-Regulation
MH  - *Gene Expression Regulation
MH  - Genetic Vectors
MH  - Humans
MH  - Immunoblotting
MH  - Inflammation
MH  - Linoleic Acids/pharmacology
MH  - Lipid Metabolism
MH  - Lipid Peroxides/pharmacology
MH  - Luciferases/metabolism
MH  - Mice
MH  - Mice, Knockout
MH  - Microscopy, Fluorescence
MH  - Molecular Sequence Data
MH  - Muscle, Smooth, Vascular/*cytology
MH  - Myocytes, Smooth Muscle/*cytology
MH  - NF-kappa B/*metabolism
MH  - Oligonucleotides/genetics
MH  - Oligonucleotides, Antisense/chemistry
MH  - Plasmids/metabolism
MH  - Polymerase Chain Reaction
MH  - Promoter Regions, Genetic
MH  - RNA Interference
MH  - RNA, Messenger/metabolism
MH  - RNA, Small Interfering/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sequence Homology, Nucleic Acid
MH  - Time Factors
MH  - Transcription, Genetic
MH  - Transfection
MH  - Up-Regulation
EDAT- 2004/04/15 05:00
MHDA- 2004/12/16 09:00
CRDT- 2004/04/15 05:00
PHST- 2004/01/23 00:00 [received]
PHST- 2004/02/10 00:00 [revised]
PHST- 2004/02/12 00:00 [accepted]
PHST- 2004/04/15 05:00 [pubmed]
PHST- 2004/12/16 09:00 [medline]
PHST- 2004/04/15 05:00 [entrez]
AID - S0022282804000483 [pii]
AID - 10.1016/j.yjmcc.2004.02.007 [doi]
PST - ppublish
SO  - J Mol Cell Cardiol. 2004 Apr;36(4):585-95. doi: 10.1016/j.yjmcc.2004.02.007.