The effects of DA-6034 on NF-kappaB activity induced by lipopolysaccharide or tumor necrosis factor alpha in a human colonic epithelial cell line.

Abstract

BACKGROUND/AIMS:

DA-6034, a synthetic derivative of eupatilin, has shown a potent anti-inflammatory effect in experimental rat models with inflammatory bowel disease (IBD). Nuclear factor kappaB (NF-kappaB) is known to play a central role in regulating inflammatory responses in patients with IBD, and is an important target for therapeutic intervention. We, therefore, examined the effects of DA-6034 on NF-kappaB activity and proinflammatory cytokines production in the human colonic cell line.

METHODOLOGY:

HT-29 cells were stimulated with either lipopolysaccharide (LPS) or tumor necrosis factor alpha (TNF-alpha). After pretreatment with different doses of DA-6034, the influence of DA-6034 on NF-kappaB activity was determined by an electrophoretic mobility shift assay (EMSA). The effect of DA-6034 on inhibitory protein kappaBalpha (IkappaBalpha) activity was determined by immunoblotting. TNF-alpha and interlukin 8 (IL-8) messenger RNA expression, which are regulated by NF-kappaB, were assayed by RT-PCR. The production of TNF-alpha or IL-8 proteins was measured by ELISA.

RESULTS:

DA-6034 inhibited NF-kappaB activity and prevented IkappaBalpha degradation in HT-29 cells after stimulation with either LPS or TNF-alpha. Furthermore, the production of TNF-alpha or IL-8 proteins decreased significantly after DA-6034 pretreatment.

CONCLUSIONS:

These observations suggest that DA-6034 produces an anti-inflammaotory effect through inhibition of NF-kappaB activity by the prevention of I B degradation.

• Authors: Ji Won Kim, Byeong Gwan Kim, Kook Lae Lee, Ji Bong Jeong, Yong Jin Jung, Joo Sung Kim, Hyun Chae Jung, In Sung Song
• Journal: Hepato-gastroenterology (Hepatogastroenterology) 2008 Nov-Dec Vol. 55 Issue 88 Pg. 2059-64 ISSN: 0172-6390 [Print] Greece
• PMID: 19260477 (Publication Type: Journal Article)

Chemical References

• Flavonoids
• Lipopolysaccharides
• NF-kappa B
• Tumor Necrosis Factor-alpha
• recoflavone

Topics

• Cell Fractionation
• Electrophoretic Mobility Shift Assay
• Flavonoids (administration & dosage, pharmacology)
• HT29 Cells
• Humans
• Lipopolysaccharides (pharmacology)
• NF-kappa B (drug effects, metabolism)
• Tumor Necrosis Factor-alpha (analysis, pharmacology)