PMID- 11901316
OWN - NLM
STAT- MEDLINE
DCOM- 20020416
LR  - 20190711
IS  - 0022-5282 (Print)
IS  - 0022-5282 (Linking)
VI  - 52
IP  - 3
DP  - 2002 Mar
TI  - Slow channel calcium inhibition blocks proinflammatory gene signaling and reduces 
      macrophage responsiveness.
PG  - 434-42
AB  - BACKGROUND: This study investigates the possible intracellular mechanisms 
      responsible for calcium antagonist protection in tissue-fixed macrophages, a 
      central modulator of the proinflammatory phenotype. METHODS: Rabbit alveolar 
      macrophages were exposed to lipopolysaccharide in the presence of different 
      specific calcium antagonists. Cellular and nuclear protein were extracted and 
      analyzed by Western blot for the phosphorylated forms of PYK2, ERK 1/2, and p38, 
      and nuclear translocation of NF-kappaB and AP-1. Tumor necrosis factor-alpha 
      (TNF-alpha) expression was measured by an L929 bioassay on cellular supernatants. 
      Statistical analysis was performed by unpaired Student's t tests. RESULTS: Cells 
      pretreated with 100 to 500 micromol/L of diltiazem or 50 to 100 micromol/L of 
      verapamil, both slow channel calcium blockers, led to dose-dependent reductions 
      in lipopolysaccharide-induced PYK2 and ERK 1/2 phosphorylation, and nuclear 
      translocation of AP-1 when compared with controls (p < 0.05). Neither inhibitor 
      had any significant effect on p38 or NF-kappaB translocation. EGTA an 
      extracellular calcium chelator, had no significant effect on any intracellular 
      process studied. A dose-dependent reduction in TNF-alpha production was 
      demonstrated with diltiazem and verapamil (p < 0.05), with no effect induced by 
      EGTA. CONCLUSION: Slow channel calcium influx is essential for optimal 
      intracellular signaling through PYK2 and ERK 1/2. This reduced intracellular 
      signaling correlated with reduced AP-1 translocation and TNF-alpha production. 
      Extracellular calcium chelation had no significant effect on intracellular 
      signaling or TNF-alpha production. This study further elucidates the protective 
      mechanism of action of calcium channel blockade by diltiazem and verapamil by 
      reducing intracellular calcium release and down-regulating the excessive 
      proinflammatory phenotype.
FAU - Cuschieri, Joseph
AU  - Cuschieri J
AD  - Department of Surgery, University of Washington, Seattle, Washington 98104, USA. 
      jcuschie@uwashington.edu
FAU - Gourlay, David
AU  - Gourlay D
FAU - Garcia, Iris
AU  - Garcia I
FAU - Jelacic, Sandra
AU  - Jelacic S
FAU - Maier, Ronald V
AU  - Maier RV
LA  - eng
GR  - GM45873-08/GM/NIGMS 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  - United States
TA  - J Trauma
JT  - The Journal of trauma
JID - 0376373
RN  - 0 (Calcium Channel Blockers)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
RN  - 0 (Transcription Factor AP-1)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - CJ0O37KU29 (Verapamil)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.2 (Focal Adhesion Kinase 2)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EE92BBP03H (Diltiazem)
SB  - IM
MH  - Analysis of Variance
MH  - Animals
MH  - Blotting, Western
MH  - Calcium Channel Blockers/*pharmacology
MH  - Diltiazem/*pharmacology
MH  - Dose-Response Relationship, Immunologic
MH  - Focal Adhesion Kinase 2
MH  - Lipopolysaccharides/pharmacology
MH  - Macrophages, Alveolar/immunology/*metabolism
MH  - Mitogen-Activated Protein Kinases/metabolism
MH  - NF-kappa B/metabolism
MH  - Protein-Tyrosine Kinases/metabolism
MH  - Rabbits
MH  - Sepsis/metabolism
MH  - Signal Transduction
MH  - Transcription Factor AP-1/metabolism
MH  - Tumor Necrosis Factor-alpha/*metabolism
MH  - Verapamil/*pharmacology
EDAT- 2002/03/20 10:00
MHDA- 2002/04/17 10:01
CRDT- 2002/03/20 10:00
PHST- 2002/03/20 10:00 [pubmed]
PHST- 2002/04/17 10:01 [medline]
PHST- 2002/03/20 10:00 [entrez]
AID - 10.1097/00005373-200203000-00004 [doi]
PST - ppublish
SO  - J Trauma. 2002 Mar;52(3):434-42. doi: 10.1097/00005373-200203000-00004.