Effect of Forsythiae Fructus Exract on the Release of Inflammatory Mediatorinduced by Lipopolysaccharide in RAW 264.7 Macrophage

  • You, Bok-Jong (Department of Oriental Medical Ophthalmology & Otolarygology & Dermatology, College of Oriental Medicine, Semyung University) ;
  • Kim, Hee-Taek (Department of Oriental Medical Ophthalmology & Otolarygology & Dermatology, College of Oriental Medicine, Semyung University)
  • Published : 2007.06.25

Abstract

Forsythiae fructus has traditionally been used for the treatment of erysipelas, skin rash and acute or chronic inflammatory disorders. The effect of Forsythiae fructus against lipopolysaccharide-induced inflammation was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), on mouse RAW 264.7 macrophages. Forsythiae fructus extract suppressed the expression of iNOS, COX-2 and NF-$_K$B mRNAs on the lipopolysaccharide-stimulated enhancement in RAW 264.7 macrophages. We examined the expression of iNOS and COX-2 in both mRNA and protein levels to investigate the mechanism by which Forsythiae fructus extract inhibits NO production. Forsythiae fructus extract significantly reduced iNOS, NF-$_K$B and PGE$_2$, but didn't inhibit COX-2 expression which was induced by LPS treatment in Raw 264.7 cells. These results suggest that Forsythiae fructus exerts anti-inflammatory effects probably by suppression of the iNOS and NF-$_K$B expressions.

Keywords

References

  1. Feng, N.P., Di, B., Liu, W.Y. Comparison of the metabolism of baicalin in rats orally administered with Radix scutellariae extract and Shuang-Huang-Lian extract. Chem Pharm Bull. 53(8):978-983, 2005 https://doi.org/10.1248/cpb.53.978
  2. Endo, K., Taguchi, T., Taguchi, F., Hikino, H., Yamahara, J., Fujimura, H. Antiinflammatory principles of Atractylodes rhizomes. Chem Pharm Bull. 27(12):2954-2958, 1979 https://doi.org/10.1248/cpb.27.2954
  3. Resch, M., Heilmann, J., Steigel, A., Bauer, R. Further phenols and polyacetylenes from the rhizomes of Atractylodes lancea and their anti-inflammatory activity. Planta Med. 67(5):437-442, 2001 https://doi.org/10.1055/s-2001-15817
  4. Yu, K.W., Kiyohara, H., Matsumoto, T., Yang, H.C., Yamada, H. Intestinal immune system modulating polysaccharides from rhizomes of Atractylodes lancea. Planta Med. 64(8):714-719, 1998 https://doi.org/10.1055/s-2006-957564
  5. Dawson, T.M., Dawson, V.L., Snyder, S.H. A novel neuronal messenger molecule in brain: the free radical, nitric oxide. Ann Neurol. 32(3):297-311, 1992 https://doi.org/10.1002/ana.410320302
  6. Yui, Y., Hattori, R., Kosuga, K., Eizawa, H., Hiki, K., Kawai, C. Purification of nitric oxide synthase from rat macrophages. J Biol Chem. 266(19):12544-12547, 1991
  7. Mitchell, J.A., Larkin, S., Williams, T,J. Cyclooxygenase-2: regulation and relevance in inflammation. Biochem Pharmacol. 50(1):1535-1542, 1995 https://doi.org/10.1016/0006-2952(95)00212-X
  8. Crofford, L.J., Lipsky, P.E., Brooks, P., Abramson, S.B., Simon, L.S., van de Putte, L.B. Basic biology and clinical application of specific cyclooxygenase-2 inhibitors. Arthritis Rheum. 43(1):4-13, 2000 https://doi.org/10.1002/1529-0131(200001)43:1<4::AID-ANR2>3.0.CO;2-V
  9. Vane, J.R., Mitchell, J.A., Appleton, I., Tomlinson, A., Bishop-Bailey, D., Croxtall, J., Willoughby, D.A. Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation. Pharmacology, 91(6):2046-2050, 1994
  10. Moncada, S., Palmer, R.M., Higgs, E.A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 43(2):109-142,1991
  11. Yun, H.Y., Dawson, V.L., Dawson, T.M. Neurobiology of nitric oxide. Crit Rev Neurobiol. 10(3-4):291-316, 1996 https://doi.org/10.1615/CritRevNeurobiol.v10.i3-4.20
  12. Funk, C.D., Funk, L.B., Kennedy, M.E., Pong, A.S., Fitzgerald, G.A. Human platelet/erythroleukemia cell prostaglandin G/H synthase: cDNA cloning, expression, and gene chromosomal assignment. FASEB J. 5(9):2304-2312, 1991
  13. Hinz, B., Brune, K. Cyclooxygenase-2--10 years later. J Pharmacol Exp Ther. 300(2):367-375, 2002 https://doi.org/10.1124/jpet.300.2.367
  14. Prescott, S.M., Fitzpatrick, F.A. Cyclooxygenase-2 and carcinogenesis, Biochim Biophys Acta. 1470(2):M69-M78, 2000
  15. Subbaramaiah, K., Dannenberg, A.J. Cyclooxygenase 2: a molecular target for cancer prevention and treatment. Trends Pharmacol Sci. 24(2):96-102, 2003 https://doi.org/10.1016/S0165-6147(02)00043-3
  16. Lappas, M., Permezel, M., Georgiou, H.M., Rice, G.E. Nuclear factor kappa B regulation of proinflammatory cytokines in human gestational tissues in vitro. Biol Reprod. 67(2):668-673, 2002 https://doi.org/10.1095/biolreprod67.2.668
  17. Surh, Y.J., Chun, K.S., Cha, H.H., Han, S.S., Keum, Y.S., Park, K.K., Lee, S.S. Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down- regulation of COX-2 and iNOS through suppression of NF-kappa B activation. Mutat Res. 243-268, 480-481, 2001
  18. Chen, F., Kuhn, D.C., Sun, S.C., Gaydos, L.J., Demers, L.M. Dependence and reversal of nitric oxide production on NF-kappa B in silica and lipopolysaccharide-induced macrophages. Biochem Biophys Res Commun. 214(3):839-846, 1995 https://doi.org/10.1006/bbrc.1995.2363
  19. Roshak, A.K., Jackson, J.R., McGough, K., Chabot-Fletcher, M., Mochan, E., Marshall, L.A. Manipulation of distinct NFkappaB proteins alters interleukin-1beta-induced human rheumatoid synovial fibroblast prostaglandin E2 formation. JJ Biol Chem. 271(49):31496-31501, 1996 https://doi.org/10.1074/jbc.271.49.31496
  20. Schmedtje, J.F. Jr, Ji, Y.S., Liu, W.L., DuBois, R.N., Runge, M.S. Hypoxia induces cyclooxygenase-2 via the NF-kappaB p65 transcription factor in human vascular endothelial cells. J Biol Chem. 272(1):601-608, 1997 https://doi.org/10.1074/jbc.272.1.601
  21. Xie, Q.W., Kashiwabara, Y., Nathan, C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J Biol Chem. 269(7):4705-4708, 1994
  22. Xie, W., Merrill, J.R., Bradshaw, W.S., Simmons, D.L. Structural determination and promoter analysis of the chicken mitogen-inducible prostaglandin G/H synthase gene and genetic mapping of the murine homolog. Arch Biochem Biophys. 300(1):247-252, 1993 https://doi.org/10.1006/abbi.1993.1034
  23. Kracht, M., Saklatvala, J. Transcriptional and post-trans- criptional control of gene expression in inflammation. Cytokine, 20(3):91-106, 2002 https://doi.org/10.1006/cyto.2002.0895
  24. Delfino, F., Walker, W.H. Hormonal regulation of the NF-kappaB signaling pathway. Mol Cell Endocrinol. 157(1-2):1-9, 1999 https://doi.org/10.1016/S0303-7207(99)00127-6
  25. Nathan, C., Xie, Q.W. Nitric oxide synthases: roles, tolls, and controls. Cell. 78(6):915-918, 1994 https://doi.org/10.1016/0092-8674(94)90266-6
  26. Vane, J.R., Mitchell, J.A., Appleton, I., Tomlinson, A., Bishop-Bailey, D., Croxtall, J., Willoughby, D.A. Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation. Proc Natl Acad Sci USA. 91(6):2046-2050, 1994
  27. Hinz, B., Brune, K., Pahl, A. Prostaglandin E(2) upregulates cyclooxygenase-2 expression in lipopolysaccharide- stimulated RAW 264.7 macrophages. Biochem Biophys Res Commun. 272(3):744-748, 2000 https://doi.org/10.1006/bbrc.2000.2859
  28. Crofford, L.J., Lipsky, P.E., Brooks, P., Abramson, S.B., Simon, L.S., van de Putte, L.B. Basic biology and clinical application of specific cyclooxygenase-2 inhibitors. Arthritis Rheum. 43(1):4-13, 2000 https://doi.org/10.1002/1529-0131(200001)43:1<4::AID-ANR2>3.0.CO;2-V
  29. Pang, L., Hoult, J.R. Cytotoxicity to macrophages of tetrandrine, an antisilicosis alkaloid, accompanied by an overproduction of prostaglandins. Biochem Pharmacol. 53(6):773-782, 1997 https://doi.org/10.1016/S0006-2952(96)00817-9
  30. Salvemini, D., Misko, T.P., Masferrer, J.L., Seibert, K., Currie, M.G., Needleman, P. Nitric oxide activates cyclooxygenase enzymes. Proc Natl Acad Sci U S A. 90(15):7240-7244, 1993
  31. Amin, A.R., Vyas, P., Attur, M., Leszczynska-Piziak, J., Patel, I.R., Weissmann, G., Abramson, S.B. The mode of action of aspirin-like drugs: effect on inducible nitric oxide synthase. Proc Natl Acad Sci U S A. 92(17):7926-7930, 1995