Planta Med 2005; 71(8): 748-753
DOI: 10.1055/s-2005-871287
Original Paper
Biochemistry and Molecular Biology
© Georg Thieme Verlag KG Stuttgart · New York

Isovitexin Suppresses Lipopolysaccharide-Mediated Inducible Nitric Oxide Synthase through Inhibition of NF-kappa B in Mouse Macrophages

Chun-Mao Lin1 , Sheng-Tung Huang2 , Yu-Chih Liang1 , Mei-Shan Lin1 , Chwen-Ming Shih1 , Yuan-Ching Chang3 , 4 , Tzong-Yueh Chen5 , Chien-Tsu Chen1
  • 1College of Medicine, Taipei Medical University, Taipei, Taiwan
  • 2Institute of Biotechnology, National Taipei University of Technology, Taipei, Taiwan
  • 3Department of General Surgery, Mackay Memorial Hospital, Taipei, Taiwan
  • 4Mackay Medicine, Nursing and Management College, Peito, Taipei, Taiwan
  • 5Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan
Further Information

Publication History

Received: October 5, 2004

Accepted: April 25, 2005

Publication Date:
11 August 2005 (online)

Abstract

Isovitexin exhibits potent antioxidant activities. In this study, the activity of nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated RAW264.7 macrophages after incubation with isovitexin was investigated. Isovitexin was able to reduce the production of hydrogen peroxide induced by LPS in mouse macrophage RAW264.7 cells. The cells incubated with isovitexin had markedly reduced LPS-stimulated NO production with an IC50 value of 58.5 μM. The expression of iNOS was also inhibited when the cells were treated with isovitexin. A transient transfection experiment showed that isovitexin suppressed the iNOS promoter and NF-κB-dependent transcriptional activities. It was also found to inhibit IKK kinase activity and prevent the degradation of IκBα in activated RAW264.7 cells. Additionally, Western blotting analysis revealed that isovitexin prevented the translocation of NF-κB from the cytoplasm to the nucleus. Our results indicate that its ROS scavenger and IKK inhibitory activities also contribute to the suppression of ROS-mediated NF-κB activity. These results suggest that isovitexin, a food phytochemical contained in dietary rice products, might have biological significance.

References

  • 1 Cuzzocrea S, Riley D P, Caputi A P, Salvemini D. Antioxidant therapy: a new pharmacological approach in shock, inflammation, and ischemia/reperfusion injury.  Pharmacol Rev. 2001;  53 135-59
  • 2 Lala P K, Chakraborty C. Role of nitric oxide in carcinogenesis and tumour progression.  Lancet Oncol. 2001;  2 149-56
  • 3 Komatsu W, Ishihara K, Murata M, Saito H, Shinohara K. Docosahexaenoic acid suppresses nitric oxide production and inducible nitric oxide synthase expression in interferon-gamma plus lipopolysaccharide-stimulated murine macrophages by inhibiting the oxidative stress.  Free Radic Biol Med. 2003;  34 1006-16
  • 4 Kang J L, Pack I S, Hong S M, Lee H S, Castranova V. Silica induces nuclear factor-kappa B activation through tyrosine phosphorylation of I kappa B-alpha in RAW264.7 macrophages.  Toxicol Appl Pharmacol. 2000;  169 59-65
  • 5 Suzuki Y J, Forman H J, Sevanian A. Oxidants as stimulators of signal transduction.  Free Radic Biol Med. 1997;  22 269-85
  • 6 Liang Y C, Huang Y T, Tsai S H, Lin-Shiau S Y, Chen C F, Lin J K. Suppression of inducible cyclooxygenase and inducible nitric oxide synthase by apigenin and related flavonoids in mouse macrophages.  Carcinogenesis. 1999;  20 1945-52
  • 7 Hayakawa M, Miyashita H, Sakamoto I, Kitagawa M, Tanaka H, Yasuda H. et al . Evidence that reactive oxygen species do not mediate NF-kappaB activation.  EMBO J. 2003;  22 3356-66
  • 8 Huang S T, Chen C T, Chieng K T, Huang S H, Chiang B H, Wang L F. et al . Inhibitory effects of a constituent from rice hull on tumor necrosis factor-α, prostaglandin E2, and cyclooxygenase-2 production in lipopolysaccharide-activated mouse macrophages.  Ann NY Acad Sci. 2005;  1042 387-95
  • 9 Ramarathnam N, Osawa T, Namiki M, Kawakishi S. Chemical studies on novel rice hull antioxidants. 2. Identification of isovitexin, a C-glycosyl flavonoid.  J Agric Food Chem. 1989;  37 316-9
  • 10 Lin C M, Chen C S, Chen C T, Liang Y C, Lin J K. Molecular modeling of flavonoids that inhibits xanthine oxidase.  Biochem Biophys Res Com. 2002;  294 167-72
  • 11 Lin C M, Chen C T, Lee H H, Lin J K. Prevention of cellular ROS damage by isovitexin and related flavonoids.  Planta Med. 2002;  68 363-5
  • 12 Lowenestein C J, Alley E W, Raval P, Snowman A M, Snyder S H, Russell S W. et al . Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon and lipopolysaccharide.  Proc Natl Acad Sci USA. 1993;  90 9730-4
  • 13 Tsai S H, Liang Y C, Lin-Shiau S Y, Lin J K. Suppression of TNFalpha-mediated NFkappaB activity by myricetin and other flavonoids through downregulating the activity of IKK in ECV304 cells.  J Cell Biochem. 1999;  74 06-15
  • 14 Kim H, Lee H S, Chang K T, Ko T H, Baek K J, Kwon N S. Chloromethyl ketones block induction of nitric oxide synthase in murine macrophages by preventing activation of nuclear factor-kB.  J Immunol. 1995;  154 4741-8
  • 15 Xie Q W, Kashiwabara Y, Nathan C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase.  J Biol Chem. 1994;  269 4705-8
  • 16 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.  Anal Biochem. 1987;  162 156-9
  • 17 Brouet I, Ohshima H. Curcumin, an anti-tumour promoter and anti-inflammatory agent, inhibits induction of nitric oxide synthase in activated macrophages.  Biochem Biophys Res Commun. 1995;  206 533-40
  • 18 Schreck R, Rieber P, Baeuerle P A. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1.  EMBO J. 1991;  10 2247-58
  • 19 Schmidt K N, Amstad P, Cerutti P, Baeuerle P A. The roles of hydrogen peroxide and superoxide as messengers in the activation of transcription factor NF-kappa B.  Chem Biol. 1995;  2 13-22
  • 20 Zhang Y, Chen F. Reactive oxygen species (ROS), troublemakers between nuclear factor-kappaB (NF-kappaB) and c-Jun NH(2)-terminal kinase (JNK).  Cancer Res. 2004;  64 1902-5

Dr. Chien-Tsu Chen

College of Medicine

Taipei Medical University

No. 250 Wu-Hsiung Street

Taipei 110

Taiwan, R.O.C.

Phone: +886-2-2736-1661 ext 2400

Fax: +886-2-2738-7348

Email: cmlin@tmu.edu.tw

    >