Translator Disclaimer
1 March 2004 AN IN VITRO SCREENING ASSAY FOR INHIBITORS OF PROINFLAMMATORY MEDIATORS IN HERBAL EXTRACTS USING HUMAN SYNOVIOCYTE CULTURES
Author Affiliations +
Abstract

Tumor necrosis factor–α (TNF-α), cyclooxygenase (COX)-2, and prostaglandin (PG)E-2 play a critical role in the pathophysiology of arthritis. Tumor necrosis factor–α 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 × 105 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-α, 10 ng/ml interleukin-1β, or control media alone at 5% carbon dioxide, 37° C. Cell viability, TNF-α, COX-2, PGE-2, nuclear factor κB (NF-κB), and inhibitory subunit I kappa B-alpha (IκB-α) expression were analyzed by reverse transcriptase–polymerase chain reaction, enzyme-linked immunosorbent assay, electrophoretic mobility shift assay, and Western blots. Ginger extract-HAPC (100 μg/ml) significantly inhibited the activation of TNF-α and COX-2 expression in human synoviocytes as well as suppressed production of TNF-α and PGE-2. Inhibition of TNF-α and COX-2 activation was accompanied by suppression of NF-κB and IκB-α 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.

CARMELITA G. FRONDOZA, AFSHIN SOHRABI, ANNA POLOTSKY, PHONG V. PHAN, DAVID S. HUNGERFORD, and LARS LINDMARK "AN IN VITRO SCREENING ASSAY FOR INHIBITORS OF PROINFLAMMATORY MEDIATORS IN HERBAL EXTRACTS USING HUMAN SYNOVIOCYTE CULTURES," In Vitro Cellular & Developmental Biology - Animal 40(3), (1 March 2004). https://doi.org/10.1290/1543-706X(2004)040<0095:AIVSAF>2.0.CO;2
Received: 7 January 2004; Accepted: 24 March 2004; Published: 1 March 2004
JOURNAL ARTICLE
7 PAGES

This article is only available to subscribers.
It is not available for individual sale.
+ SAVE TO MY LIBRARY

SHARE
ARTICLE IMPACT
RIGHTS & PERMISSIONS
Get copyright permission
Back to Top