A Sesquiterpene Lactone, Costunolide, from Magnolia grandiflora Inhibits NF-κB by Targeting IκB Phosphorylation

Abstract

A sesquiterpene lactone, costunolide (CTN), was identified from Magnolia grandiflora together with parthenolide (PTN) by its strong inhibition of LPS-induced NF-κB activation. CTN, which showed more potent inhibition than PTN in the NF-κB activation, strongly suppressed nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. RT-PCR and Western blot analyses demonstrated that CTN suppressed the expression of iNOS mRNA and protein in a dose-dependent manner. CTN also significantly inhibited LPS-induced DNA-binding activity of NF-κB as well as the LPS-induced degradation of IκB-α and -β. Furthermore, CTN inhibited LPS-induced phosphorylation of IκB-α. These findings support that CTN inhibits NO production by down-regulating iNOS expression, at least, in part through the inhibition of IκBs' phosphorylation and degradation, which are essential for the activation of NF-κB.

Magnolia grandiflora - Magnoliaceae - costunolide - inducible nitric oxide synthase - NF-κB - IκB

References

• 1 Baldwin A S. The NF-kappaB and I kappa B proteins: new discoveries and insights.  Annual Review of Immunology. 1996;  14 649-83
• 2 Vane J R. The Croonian Lecture, 1993. The endothelium: maestro of the blood circulation.  Philosophical Transactions of Royal Society of London Series B, Biological Sciences. 1994;  343 225-46
• 3 Dinerman J L, Lowenstein C J, Snyder S H. Molecular mechanisms of nitric oxide regulation. Potential relevance to cardiovascular disease.  Circulation Research. 1993;  73 217-22
• 4 Xie Q, Nathan C. The high-output nitric oxide pathway: role and regulation.  Journal of Leukocyte Biology. 1994;  56 576-82
• 5 Xie Q W, Whisnant R, Nathan C. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide.  Journal of Experimental Medicine. 1993;  177 1779-84
• 6 Xie Q W, Kashiwabara Y, Nathan C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase.  Journal of Biological Chemistry. 1994;  269 4705-8
• 7 Baeuerle P A, Henkel T. Function and activation of NF-kappa B in the immune system.  Annual Review of Immunology. 1994;  12 141-79
• 8 El-Feraly F S, Chan Y M. Isolation and characterization of the sesquiterpene lactones costunolide, parthenolide, costunolide diepoxide, santamarine, and reynosin from Magnolia grandiflora L.  Journal of Pharmaceutical Science. 1978;  67 347-50
• 9 Erkel G, Anke T, Sterner O. Inhibition of NF-kappa B activation by panepoxydone.  Biochemical and Biophysical Research Communications. 1996;  226 214-21
• 10 Cullen B R, Malim M H. Secreted placental alkaline phosphatase as an eukaryotic reporter gene.  Methods in Enzymology. 1992;  216 362-8
• 11 Gilad E, Wong H R, Zingarelli B, Virag L, O'Connor M, Salzman A L, Szabo C. Melatonin inhibits expression of the inducible isoform of nitric oxide synthase in murine macrophages: role of inhibition of NFkappaB activation.  FASEB Journal. 1998;  12 685-93
• 12 Schmidt  HHHW, Kelm M. Determination of nitrite and nitrate by the Griess reaction.  In: Feelisch M, Stamler J, editors. Methods in nitric oxide research, John Wiley & Sons Ltd., New York. 1996;  Chapter 33 491-7
• 13 Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.  Journal of Immunological Methods. 1983;  65 55-63
• 14 Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Analytical Biochemistry. 1976;  72 248-54
• 15 Groenewegen W A, Heptinstall S. A comparison of the effects of an extract of feverfew and parthenolide, a component of feverfew, on human platelet activity in-vitro .  Journal of Pharmacy and Pharmacology. 1990;  42 553-7
• 16 Hwang D, Fischer N H, Jang B C, Tak H, Kim J K, Lee W. Inhibition of the expression of inducible cyclooxygenase and proinflammatory cytokines by sesquiterpene lactones in macrophages correlates with the inhibition of MAP kinases.  Biochemical Biophysical Research Communications. 1996;  226 810-8
• 17 Sumner H, Salan U, Knight D W, Hoult J R. Inhibition of 5-lipoxygenase and cyclooxygenase in leukocytes by feverfew.  Involvement of sesquiterpene lactones and other components.. Biochemical Pharmacology. 1992;  43 2313-20
• 18 Hehner S P, Heinrich M, Bork P M, Vogt M, Ratter F, Lehmann V, Schulze-Osthoff K, Dröge W, Schmitz M L. Sesquiterpene lactones specifically inhibit activation of NF-kappa B by preventing the degradation of I kappa B-alpha and I kappa B-beta.  Journal of Biological Chemistry. 1998;  273 1288-97
• 19 Wong H R, Menendez I Y. Sesquiterpene lactones inhibit inducible nitric oxide synthase gene expression in cultured rat aortic smooth muscle cells.  Biochemical Biophysical Research Communications. 1999;  262 375-80
• 20 Hehner S P, Hofmann T G, Dröge W, Schmitz M L. The anti-inflammatory sesquiterpene lactone parthenolide inhibits NF-kappa B by targeting the I kappa B kinase complex.  Journal of Immunology. 1999;  163 5617-23
• 21 Lyss G, Knorre A, Schmidt T J, Pahl H L, Merfort I. The anti-inflammatory sesquiterpene lactone helenalin inhibits the transcription factor NF-kappaB by directly targeting p65.  Journal of Biological Chemistry. 1998;  273 33508-16
• 22 Kopp E, Ghosh S. Inhibition of NF-kappa B by sodium salicylate and aspirin.  Science. 1994;  265 956-9
• 23 Auphan N, DiDonato J A, Rosette C, Helmberg A, Karin M. Immunosuppression by glucocorticoids: inhibition of NF-kappa B activity through induction of I kappa B synthesis.  Science. 1995;  270 286-90