Sauchinone, a Lignan from Saururus chinensis, Suppresses iNOS Expression through the Inhibition of Transactivation Activity of RelA of NF-κB
Bang Yeon Hwang1,2
, Jeong-Hyung Lee1
, Haeng Sun Jung1,3
, Kyung-Sook Kim1
, Jeong Beom Nam1
, Young Soo Hong1
, Sang-Gi Paik3
, Jung Joon Lee1
1Anticancer Agents Research Laboratory, Korea Research Institute of Bioscience and Biotechnology, Yuseong, Daejeon, Korea
2College of Pharmacy, Chungbuk National Univeristy, Cheongju, Korea
3Department of Biology, Chungnam National University, Daejeon, Korea
This work was supported in part by a research grant (PF0320701-00) from Plant Diversity Research Center of 21st Frontier Research Program funded by the Korean Ministry of Science and Technology and by a grant from KRIBB Research Initiative Program
Sauchinone, a known lignan, was isolated from the root of Saururus chinensis as an active principle responsible for inhibiting the production of NO in LPS-stimulated RAW264.7 cells by activity-guided fractionation. Sauchinone dose-dependently inhibited not only the production of NO, but also the expression of iNOS mRNA and protein in LPS-stimulated RAW 264.7 cells. Furthermore, sauchinone prevented LPS-induced NF-κB activation, which is known to play a critical role in iNOS expression, assessed by a reporter assay under the control of NF-κB. However, an electrophoretic mobility shift assay (EMSA) demonstrated that sauchinone did not suppress the DNA-binding activity of NF-κB or the degradation of IκB-α induced by LPS. Further analysis revealed that transactivation activity of RelA subunit of NF-κB was dose-dependently suppressed in the presence of sauchinone. Taken together, our results suggested that sauchinone could inhibit production of NO in LPS-stimulated RAW264.7 cells through the suppression of NF-κB by inhibiting transactivation activity of RelA subunit.
3
Goldring C E, Reveneau S, Algarte M, Jeannin J F.
In vivo footprinting of the mouse inducible nitric oxide synthase gene: inducible protein occupation of numerous sites including Oct and NF-IL6.
Nucleic Acids Res.
1996;
24
1682-7
8
Mercurio F, Zhu H, Murray B W, Shevchenko A, Bennett B L, Li J, Young D B, Barbosa M, Mann M, Manning A, Rao A.
IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for NF-kappaB activation.
Science.
1997;
278
860-6
12
Hwang B Y, Lee J H, Koo T H, Kim H S, Hong Y S, Ro J S, Lee K S, Lee J J.
Kaurane diterpenes from Isodon japonicus inhibit nitric oxide and prostaglandin E2 production and NF-kappaB activation in LPS-stimulated macrophage RAW264.7 cells.
Planta Med.
2001;
67
406-10
13 Schmidt H HHW, Kelm M. Determination of nitrite and nitrate by the Griess reaction. In: Methods in nitric oxide research. London; John Wiley & Sons Ltd 1996: p. 491-7
14
Scudiero D A, Shoemaker R H, Paull K D, Monks A, Tierney S, Nofziger T H, Currens M J, Seniff D, Boyd M R.
Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines.
Cancer Res.
1988;
48
4827-33
15
Lee J H, Koo T H, Hwang B Y, Lee J J.
Kaurane diterpene, kamebakaurin, inhibits NF-κB by directly targeting the DNA-binding activity of p50 and blocks the expression of antiapoptotic NF-κB target genes.
J Biol Chem.
2002;
277
18 411-20
16
Koo T H, Lee J H, Park Y J, Hong Y S, Jim HS Kim K W, Lee J J.
A sesquiterpene lactone, costunolide, from Magnolia grandiflora inhibits NF-kB by targeting IkB phosphorylation.
Planta Med.
2001;
67
103-7
17
Lee A K, Sung S H, Kim Y C, Kim S G.
Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-α and COX-2 expression by sauchinone effects on I-κBα phosphorylation, C/EBP and AP-1 activation.
Br J Pharmacol.
2003;
139
11-20
18
Vermeulen L, De Wilde G, Notebaert S, Vanden Berghe W, Haegeman G.
Regulation of the transcriptional activity of the nuclear factor-kappaB p65 subunit.
Biochem Pharmacol.
2002;
64
963-70
19
Zhong H, May M J, Jimi E, Ghosh S.
The phosphorylation status of nuclear NF-kappa B determines its association with CBP/p300 or HDAC-1.
Mol Cell.
2002;
9
625-36
20
Lee K Y, Chang W, Qiu D, Kao P N, Rosen G D.
PG490 (triptolide) cooperates with tumor necrosis factor-α to induce apoptosis in tumor cells.
J Biol Chem.
1999;
274
13 451-5