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Inhibition of Lipopolysaccharide-Induced iNOS, COX- 2, and TNF-α Expression by Aqueous Extract of Orixa Japonica in RAW 264.7 Cells via Suppression of NF- <sub>k</sub>B Activity
Abstract
Methods: The expression of mRNA and protein using RT-PCR and Western blot analysis was investigated. The level of nitric oxide (NO) production was analyzed using Griess reaction. Release of prostaglandin E2 (PGE2) and tumor necrosis factor- (TNF-) was determined using sandwich ELISA.
Results: AEOJ potently inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor- (TNF-) in LPS-stimulated RAW 264.7 cells. Consistent with these findings, AEOJ was
also found to significantly reduce LPS-induced expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF- at the transcriptional level. Additionally, AEOJ attenuated LPSinduced
NF-B activity via the inhibition of IB phosphorylation and degradation. It was also found that the NF-B inhibitor N-acetyl cysteine (NAC) attenuated LPS-induced gene expression of iNOS, COX-2,
and TNF-. These results indicate that AEOJ attenuates LPS-induced inflammatory mediators such as NO, PGE2, and TNF- via suppression of NF-B activity.
Conclusion: These results suggest that AEOJ has a potential activity to alleviate LPS-induced inflammation.
