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


METHODS
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.
Kim
Ji Won
JW
Department of Internal Medicine, Seoul Metropolitan Boramae Hospital, Seoul, Korea.
Kim
Byeong Gwan
BG
Lee
Kook Lae
KL
Jeong
Ji Bong
JB
Jung
Yong Jin
YJ
Kim
Joo Sung
JS
Jung
Hyun Chae
HC
Song
In Sung
IS
D002458:Cell Fractionation; D024202:Electrophoretic Mobility Shift Assay; D005419:Flavonoids / Q000008:administration & dosage / Q000494:pharmacology*; D019073:HT29 Cells; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D016328:NF-kappa B / Q000187:drug effects* / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000494:pharmacology
D005419:Flavonoids; D008070:Lipopolysaccharides; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; C121889:recoflavone