An in Vitro Screening Assay for Inhibitors of Proinflammatory Mediators in Herbal Extracts Using Human Synoviocyte Cultures

Abstract

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.

Zingiber officinale(Ginger): A Future Outlook on Its Potential in Prevention and Treatment of Diabetes and Prediabetic States

“…To investigate the chemical component that may be responsible for the anti-inflammatory activities we investigated the effects of the major component (S)- [6]-gingerol on inflammatory pathways in hepatocytes. Our studies and other studies in the literature have shown that ginger extracts suppress inflammation through inhibition of the classical nuclear factor-kappa B (NF-B) pathway in various cell types and tissues [42,47]. Upon exposure to proinflammatory stimuli, activated NF-B directs transcription of target genes [45], including genes encoding cytokines, chemokines, and the enzyme cyclooxygenase 2 (COX2) [48].…”

Use ofCordia dichotomabark in the treatment of ulcerative colitis

“…On the other hand, the methanol fraction of C. dichotoma bark decreased significantly the gross lesion scores, and production of both TNF-α and PGE 2 . The effect of the methanol fraction on TNF-α may be mediated either by blocking activation of this pro-inflammatory mediator and its transcriptional regulator (Frondoza et al, 2004), and/or inhibits its production from macrophages (Tripathi et al, 2007). Inhibition of PGE 2 , on the other hand, may follow that of TNF-α (Otani et al, 2006), or may result from its ability to inhibit cycloxygenase enzymes (Grzanna et al, 2005).…”

Side-Effects of Non-Steroidal Anti-Inflammatory Drugs on the Liver in Dogs and Hepatoprotective Effect of Plant Remedies

“…In human medicine herb hyssop and icelandic lichen are indicated for the treatment of cholelithiasis and cholestasis, thyme herb helps digestion of fat and acts relaxant on the bile ducts, while the saponariae root increases the amount of bile secreted by the liver and facilitate its flow into the duodenum (2,5,14,19). Our study for the first time concerns the relationship between components of Pectosol® and liver pathology.…”

“…Pectosol® (composition: saponariae root, herb hyssop, icelandic lichen, thyme herb, extraction solvent 70% etanole and 10 g glycerol) has been used successfully in human medicine to protect the liver during NSAID therapy (14,19). This remedy contain many bioactive compounds with potentially hepatoprotective effects.…”