PMID- 10792620
OWN - NLM
STAT- MEDLINE
DCOM- 20000601
LR  - 20081121
IS  - 0085-2538 (Print)
IS  - 0085-2538 (Linking)
VI  - 57
IP  - 5
DP  - 2000 May
TI  - Lipopolysaccharide-induced MCP-1 gene expression in rat tubular epithelial cells 
      is nuclear factor-kappaB dependent.
PG  - 2011-22
AB  - BACKGROUND: Endotoxin is an important factor in the development of acute renal 
      failure related to infection and in acceleration of chronic nephritis. 
      Lipopolysaccharide (LPS; the major component of endotoxin) is one of the most 
      potent triggers for renal cells to produce monocyte chemoattractant protein-1 
      (MCP-1), a key cytokine involved in immune cell recruitment into the renal 
      interstitium in acute and chronic renal diseases. Knowledge about the 
      transcriptional regulation of MCP-1 in renal tubular epithelial cells in response 
      to LPS is incomplete. METHODS: Transcriptional regulation of MCP-1 was 
      investigated in rat proximal tubule cells (PTCs) in primary culture and was 
      exposed to LPS using electromobility shift assay and supershift analysis for 
      nuclear factor-kappaB (NF-kappaB) and Western blot for the NF-kappaB inhibitory 
      protein IkappaB. To prove the role for NF-kappaB, activator protein (AP-1), and 
      sequence-specific transcription factor (Sp1), mutation and deletion analysis was 
      performed using a 3.5 kb fragment of rat MCP-1 5'-flanking region inserted into a 
      luciferase reporter construct transfected into tubular epithelial cell line 
      (NRK-52E). RESULTS: LPS increased NF-kappaB in a dose- and time-dependent manner, 
      which paralleled that of MCP-1 mRNA expression. IkappaBalpha decreased within 30 
      minutes of LPS treatment, but returned to basal levels by two hours. IkappaBbeta 
      levels were depressed within one hour and remained low throughout the culture 
      period after LPS stimulation. The activity of the transfected 5'-flanking region 
      of the MCP-1 gene increased nearly threefold after LPS stimulation. Mutation or 
      deletion of NF-kappaB binding sites, located in the enhancer region of the 
      5'-flanking region, resulted in a total loss of LPS-induced increase in 
      luciferase activity. Mutation of putative AP-1 and Sp1 sites, located in the 
      proximal promoter region of MCP-1, reduced basal luciferase activity in 
      unstimulated cells, but had no effect on LPS-stimulated luciferase activity. 
      CONCLUSIONS: These studies prove that NF-kappaB is critical for LPS-induced MCP-1 
      transcription, and AP-1 and Sp1 are essential for basal expression of MCP-1 in 
      rat tubule cells. The species-specific nature of transcriptional regulation of 
      MCP-1 has important implications for the delineation of treatment to prevent 
      endotoxin-mediated renal injury.
FAU - Wang, Y
AU  - Wang Y
AD  - Department of Renal Medicine and Storr Liver Unit, The University of Sydney at 
      Westmead Hospital, Sydney, New South Wales, Australia.
FAU - Rangan, G K
AU  - Rangan GK
FAU - Goodwin, B
AU  - Goodwin B
FAU - Tay, Y C
AU  - Tay YC
FAU - Harris, D C
AU  - Harris DC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Kidney Int
JT  - Kidney international
JID - 0323470
RN  - 0 (Chemokine CCL2)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
RN  - 0 (Sp1 Transcription Factor)
RN  - 0 (Transcription Factor AP-1)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Cells, Cultured
MH  - Chemokine CCL2/*genetics
MH  - Epithelial Cells/drug effects/metabolism
MH  - Gene Expression Regulation/*drug effects
MH  - Kidney Tubules/*drug effects/metabolism
MH  - Lipopolysaccharides/*pharmacology
MH  - Male
MH  - Molecular Sequence Data
MH  - NF-kappa B/*physiology
MH  - Promoter Regions, Genetic
MH  - Rats
MH  - Rats, Wistar
MH  - Sp1 Transcription Factor/physiology
MH  - Transcription Factor AP-1/physiology
EDAT- 2000/05/03 09:00
MHDA- 2000/06/03 09:00
CRDT- 2000/05/03 09:00
PHST- 2000/05/03 09:00 [pubmed]
PHST- 2000/06/03 09:00 [medline]
PHST- 2000/05/03 09:00 [entrez]
AID - S0085-2538(15)46953-4 [pii]
AID - 10.1046/j.1523-1755.2000.00051.x [doi]
PST - ppublish
SO  - Kidney Int. 2000 May;57(5):2011-22. doi: 10.1046/j.1523-1755.2000.00051.x.