PMID- 15311968
OWN - NLM
STAT- MEDLINE
DCOM- 20041012
LR  - 20240109
IS  - 1071-2690 (Print)
IS  - 1071-2690 (Linking)
VI  - 40
IP  - 3-4
DP  - 2004 Mar-Apr
TI  - An in vitro screening assay for inhibitors of proinflammatory mediators in herbal 
      extracts using human synoviocyte cultures.
PG  - 95-101
AB  - Tumor necrosis factor-alpha (TNF-alpha), cyclooxygenase (COX)-2, and 
      prostaglandin (PG)E-2 play a critical role in the pathophysiology of arthritis. 
      Tumor necrosis factor-alpha mediates induction of other cytokines, COX-2, PGs, 
      and metalloproteinases, which leads to cartilage degradation. We developed an in 
      vitro human synoviocyte assay system for screening inhibitors of proinflammatory 
      mediators in herbal extracts. Synoviocytes (5 x 10(5) cells/well) obtained during 
      primary knee replacement from osteoarthritic patients were incubated with: 
      control media alone or ginger extract (hydroxy-methoxy-phenyl compounds [HAPC]: 
      EV.EXT 77), 1 h before activation with 1 ng/ml TNF-alpha, 10 ng/ml 
      interleukin-1beta, or control media alone at 5% carbon dioxide, 37 degrees C. 
      Cell viability, TNF-alpha, COX-2, PGE-2, nuclear factor kappaB (NF-kappaB), and 
      inhibitory subunit I kappa B-alpha (IkappaB-alpha) expression were analyzed by 
      reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent 
      assay, electrophoretic mobility shift assay, and Western blots. Ginger 
      extract-HAPC (100 microg/ml) significantly inhibited the activation of TNF-alpha 
      and COX-2 expression in human synoviocytes as well as suppressed production of 
      TNF-alpha and PGE-2. Inhibition of TNF-alpha and COX-2 activation was accompanied 
      by suppression of NF-kappaB and IkappaB-alpha induction. Using our in vitro 
      assay, we discovered that the ginger extract blocks activation of proinflammatory 
      mediators and its transcriptional regulator suggesting its mode of action. These 
      observations indicate that ginger extract-HAPC offers a complementary and 
      alternative approach to modulate the inflammatory process involved in arthritis.
FAU - Frondoza, Carmelita G
AU  - Frondoza CG
AD  - Division of Arthritis Surgery, Department of Orthopaedic Surgery, Johns Hopkins 
      University, Baltimore, Maryland 21205, USA. cgfrondo@jhmi.edu
FAU - Sohrabi, Afshin
AU  - Sohrabi A
FAU - Polotsky, Anna
AU  - Polotsky A
FAU - Phan, Phong V
AU  - Phan PV
FAU - Hungerford, David S
AU  - Hungerford DS
FAU - Lindmark, Lars
AU  - Lindmark L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - In Vitro Cell Dev Biol Anim
JT  - In vitro cellular & developmental biology. Animal
JID - 9418515
RN  - 0 (Cyclooxygenase 2 Inhibitors)
RN  - 0 (Cyclooxygenase Inhibitors)
RN  - 0 (I-kappa B Proteins)
RN  - 0 (Inflammation Mediators)
RN  - 0 (Interleukin-1)
RN  - 0 (Isoenzymes)
RN  - 0 (Membrane Proteins)
RN  - 0 (NF-kappa B)
RN  - 0 (NFKBIA protein, human)
RN  - 0 (Plant Extracts)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 139874-52-5 (NF-KappaB Inhibitor alpha)
RN  - EC 1.14.99.1 (Cyclooxygenase 2)
RN  - EC 1.14.99.1 (PTGS2 protein, human)
RN  - EC 1.14.99.1 (Prostaglandin-Endoperoxide Synthases)
RN  - K7Q1JQR04M (Dinoprostone)
SB  - IM
MH  - Blotting, Western
MH  - Cell Survival
MH  - Cells, Cultured
MH  - Cyclooxygenase 2
MH  - Cyclooxygenase 2 Inhibitors
MH  - Cyclooxygenase Inhibitors/pharmacology
MH  - Dinoprostone/metabolism
MH  - Drug Evaluation, Preclinical
MH  - Electrophoretic Mobility Shift Assay
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Gene Expression Regulation/drug effects
MH  - *Zingiber officinale
MH  - Humans
MH  - I-kappa B Proteins/metabolism
MH  - Inflammation Mediators/*antagonists & inhibitors
MH  - Interleukin-1/*antagonists & inhibitors/pharmacology
MH  - Isoenzymes/metabolism
MH  - Membrane Proteins
MH  - NF-KappaB Inhibitor alpha
MH  - NF-kappa B/analysis/metabolism
MH  - Osteoarthritis/*pathology
MH  - Plant Extracts/*pharmacology
MH  - Prostaglandin-Endoperoxide Synthases/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Synovial Membrane/*cytology/drug effects/metabolism
MH  - Tumor Necrosis Factor-alpha/*antagonists & inhibitors/pharmacology
EDAT- 2004/08/18 05:00
MHDA- 2004/10/13 09:00
CRDT- 2004/08/18 05:00
PHST- 2004/08/18 05:00 [pubmed]
PHST- 2004/10/13 09:00 [medline]
PHST- 2004/08/18 05:00 [entrez]
AID - 0401001 [pii]
AID - 10.1290/1543-706x(2004)040<0095:aivsaf>2.0.co;2 [doi]
PST - ppublish
SO  - In Vitro Cell Dev Biol Anim. 2004 Mar-Apr;40(3-4):95-101. doi: 
      10.1290/1543-706x(2004)040<0095:aivsaf>2.0.co;2.