Subscribe to RSS
DOI: 10.1055/s-2003-45189
© Georg Thieme Verlag Stuttgart · New York
Sauchinone, a Lignan from Saururus chinensis, Suppresses iNOS Expression through the Inhibition of Transactivation Activity of RelA of NF-κB
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 ProgramPublication History
Received: April 25, 2003
Accepted: September 28, 2003
Publication Date:
29 January 2004 (online)
Abstract
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.
Key words
Saururus chinensis - Saururaceae - sauchinone - inducible nitric oxide synthase - NF-κB - transactivation - RelA
References
- 1 Anggard E. Nitric oxide: mediator, murderer, and medicine. Lancet. 1994; 343 1199-206
- 2 Hobbs A J, Higgs A, Moncada S. Inhibition of nitric oxide synthase as a potential therapeutic target. Annu Rev Pharmacol Toxicol. 1999; 39 191-220
- 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
- 4 Ghosh S, May M J, Kopp E B. NF-κB and Rel proteins: Evolutionarily conserved mediators of immune responses. Annu Rev Immunol. 1998; 16 225-60
- 5 Baeuerle P A, Baltimore D. A 65-KD subunit of active NF-κB is required for inhibition of NF-κB by IκB. Genes Dev. 1989; 3 1689-98
- 6 Schmitz M L, Baeuerle P A. The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B. EMBO J. 1991; 10 3805-17
- 7 Baldwin AS J r. The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol. 1996; 14 649-83
- 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
- 9 Kim J G. Encyclopedia of illustrated Korean natural drugs. Seoul; Namsandang 1984: p. 174
- 10 Sung S H, Kim Y C. Hepatoprotective diastereomeric lignans from Saururus chinensis herbs. J Nat Prod. 2000; 63 1019-21
- 11 Wang E C, Shih M H, Liu M C, Chen M T, Lee G H. Studies of constituents of Saururus chinensis . Heterocycles. 1996; 43 969-76
- 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
Dr. Jung Joon Lee
Anticancer Agents Research Laboratory
Korea Research Institute of Bioscience and Biotechnology
P.O. Box 115
Yuseong
Daejeon 305-600
Korea
Phone: +82-42-860-4360
Fax: +82-42-860-4595
Email: jjlee@kribb.re.kr