DOI QR코드

DOI QR Code

Effects of Isatidis Radix and it's Active Component, Tryptanthrin on the Production of Inflammatory Mediators in Lipopolysaccharide-activated Raw264.7 Cells

LPS로 활성화된 Raw264.7 cell에서 판람근 및 Tryptanthrin의 염증매개물질억제효과

  • Park, Sook-Jahr (College of Oriental Medicine, Daegu Haany University) ;
  • Lee, Jong-Rok (College of Oriental Medicine, Daegu Haany University) ;
  • Jo, Mi-Jeong (College of Oriental Medicine, Daegu Haany University) ;
  • Park, Sang-Mi (College of Oriental Medicine, Daegu Haany University) ;
  • Buyn, Sung-Hui (College of Oriental Medicine, Daegu Haany University) ;
  • Cho, Il-Je (College of Oriental Medicine, Daegu Haany University) ;
  • Kim, Sang-Chan (College of Oriental Medicine, Daegu Haany University)
  • Received : 2011.03.08
  • Accepted : 2011.04.01
  • Published : 2011.04.25

Abstract

Objectives : 판람근(板藍根)은 십자화과에 속하는 대청(大靑) 또는 숭남의 근(根)을 건조한 것이다. 본 연구는 판람근(板藍根)이 청열해독(淸熱解毒)함에 근거하여, LPS로 활성화된 Raw264.7 cell에서 판람근(板藍根)과 그 성분중의 하나인 tryptanthrin이 염증매개물질에 미치는 효과를 살펴보고자 하였다. Methods : 세포생존율은 MTT, nitric oxide (NO)는 Griess reagent를 사용하여 측정하였으며, 각 단백질의 발현량은 Western blot 방법을 사용하였으며, cytokine 및 cyclooxygenase-2 (COX-2)는 ELISA방법을 사용하여 측정하였다. Results : LPS는 NO 및 prostaglandin E2 (PGE2)를 유의하게 상승시켰으며, 판람근(板藍根)추출물 (IRE) 및 tryptanthrin 은 이들을 유의하게 억제하였다. 그러나 판람근(板藍根)의 또 다른 성분인 indigo는 유의한 결과를 나타내지 못하였다. IRE와 tryptanthrin은 inhibitory kappa B alpha의 인산화를 억제하여, nuclear factor-${\kappa}$B (NF-${\kappa}$B)의 핵으로의 전위(轉位)를 억제하여, iNOS 및 cytokine을 억제하였다. IRE와 tryptanthrin의 PGE2 억제는, COX-2의 발현억제에서가 아니라, COX-2의 활성을 억제함에서 기인하였다. Conclusion : 이러한 결과는 판람근(板藍根)이 NF-${\kappa}$B pathway를 경유하여 iNOS의 발현 및 COX-2의 활성을 억제함을 나타내며, 이러한 판람근(板藍根)의 항염증효능은 일부 tryptanthrin의 작용에서 기인함을 시사한다.

Keywords

References

  1. Fitzpatrick FA. Inflammation, carcinogenesis and cancer. Int. Immunopharmacol. 2001;1:1651-67. https://doi.org/10.1016/S1567-5769(01)00102-3
  2. Chen H. Cellular inflammatory responses: novel insights for obesity and insulin resistance. Pharmacol. Res. 2006;53:469-77. https://doi.org/10.1016/j.phrs.2006.03.003
  3. Murakami A, Ohigashi H. Targeting NOX, INOS and COX-2 in inflammatory cells: chemoprevention using food phytochemicals. Int. J. Cancer. 2007;121:2357-63. https://doi.org/10.1002/ijc.23161
  4. Mariotto S, Suzuki Y, Persichini T, Colasanti M, Suzuki H, Cantoni O. Cross-talk between NO and arachidonic acid in inflammation. Curr. Med. Chem. 2007;14;1940-4. https://doi.org/10.2174/092986707781368531
  5. Lee AK, Sung SH, Kim YC, Kim SG. Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-$\alpha$ and COX-2 expression by sauchinone effects on I-$\kappa$B $\alpha$ phosphorylation, C/EBP and AP-1 activation. Br. J. Pharmacol. 2003;139:1-20. https://doi.org/10.1038/sj.bjp.0705217
  6. Tome L, Yu L, de Castro I, Campos S, Seguro A. Beneficial and harmful effects of L-arginine on renal ischaemia. Nephrol. Dial. Transplant. 1999;14:1139-45. https://doi.org/10.1093/ndt/14.5.1139
  7. Hinson RM, Williams JA, Shacter E. Elevated interleukin 6 is induced by prostaglandin E2 in a murine model of inflammation: possible role of cyclooxygenase-2. Proc. Natl. Acad. Sci. USA. 1996;93:4885-90. https://doi.org/10.1073/pnas.93.10.4885
  8. Chen F, Castranova V, Shi X. New insight into the role of nuclear factor-$\kappa$B in cell growth regulation. Am J Pathol. 2001;159(2):387-97. https://doi.org/10.1016/S0002-9440(10)61708-7
  9. Li Q, Verma IM. NF-kappaB regulation in the immune system. Nat. Rev. Immunol.. 2002;2:725-34. https://doi.org/10.1038/nri910
  10. Van Waes C. Nuclear factor-kappaB in development, prevention, and therapy of cancer. Clin. Cancer Res. 2007;13:1076-82. https://doi.org/10.1158/1078-0432.CCR-06-2221
  11. Zhang E. "The Chinese materia medica," Publishing house of Shanghai University of traditional Chinese medicine, Shanghai. 2000.
  12. Chen ZW, Wu LW, Liu ST, Cai CP, Rao PF, Ke LJ. Mechanism study of anti-influenza effects of Radix Isatidis water extract by red blood cells capillary electrophoresis. Zhongguo Zhong Yao Za Zhi. 2006;31(20):1715-9.
  13. Lin CW, Tsai FJ, Tsai CH, Lai CC, Wan L, Ho TY, Hsieh CC, Chao PD. Anti-SARS coronavirus 3C-like protease effects of Isatis indigotica root and plant-derived phenolic compounds. Antiviral Res. 2005;68:36-42. https://doi.org/10.1016/j.antiviral.2005.07.002
  14. Liu YH, Liu YF, Guo XX. Current studies on anti-endotoxic chemical components of traditional Chinese medicine in China. Acta Pharmacol. Sin. 2001;22:1071-7.
  15. Zhao YL, Wang JB, Shan LM, Jin C, Ma L, Xiao XH. Effect of Radix isatidis polysaccharides on immunological function and expression of immune related cytokines in mice. Chin. J. Integr. Med. 2008;14:207-11. https://doi.org/10.1007/s11655-008-0207-2
  16. Kong W, Zhao W, Zhao Y, Shan L, Xiao X, Guo W. Thermochemical Studies on the Quantity-Antibacterial Effect Relationship of Four Organic Acids from Radix Isatidis on Escherichia coli Growth. Biol. Pharm. Bull. 2008;31:1301-5. https://doi.org/10.1248/bpb.31.1301
  17. Fang JG, Liu YH, Wang WQ, Xie W, Fang SX, Han HG. The anti-endotoxic effect of o-aminobenzoic acid from Radix Isatidis. Acta Pharmacol. Sin. 2005;26:593-7. https://doi.org/10.1111/j.1745-7254.2005.00080.x
  18. Liau BC, Jong TT, Lee MR, Chen SS. LC-APCI-MS method for detection and analysis of tryptanthrin, indigo, and indirubin in daqingye and banlangen. J. Pharm. Biomed. Anal. 2007;43:346-51. https://doi.org/10.1016/j.jpba.2006.06.029
  19. Delgado AV, McManus AT, Chambers JP. Production of tumor necrosis factor-alpha, interleukin 1-beta, interleukin 2, and interleukin 6 by rat leukocyte subpopulations after exposure to substance P. Neuropeptides. 2003;37:355-61. https://doi.org/10.1016/j.npep.2003.09.005
  20. Danz H, Stoyanova S, Wippich P, Brattstrom A, Hamburger M. Identification and isolation of the cyclooxygenase-2 inhibitory principle in Isatis tinctoria. Planta Med. 2001;67:411-6. https://doi.org/10.1055/s-2001-15805
  21. Oberthur C, Jaggi R, Hamburger M. HPLC based activity profiling for 5-lipoxygenase inhibitory activity in Isatis tinctoria leaf extracts. Fitoterapia. 2005;76:324-32. https://doi.org/10.1016/j.fitote.2005.03.002
  22. Ishihara T, Kohno K, Ushio S, Iwaki K, Ikeda M, Kurimoto M. Tryptanthrin inhibits nitric oxide and prostaglandin E(2) synthesis by murine macrophages. Eur. J. Pharmacol. 2000;407:197-204. https://doi.org/10.1016/S0014-2999(00)00674-9
  23. Adachi J, Mori Y, Matsui S, Takigami H, Fujino J, Kitagawa H, Miller III CA, Kato T, Saeki K, Matsuda T. Indirubin and indigo are potent aryl hydrocarbon receptor ligands present in human urine. J. Biol. Chem, 2001;276:31475-8. https://doi.org/10.1074/jbc.C100238200
  24. Farias-Silva E, Cola M, Calvo TR, Barbastefano V, Ferreira AL, De Paula Michelatto D, Alves de Almeida AC, Hiruma-Lima CA, Vilegas W, Brito AR. Antioxidant activity of indigo and its preventive effect against ethanol-induced DNA damage in rat gastric mucosa. Planta Med. 2007;73:1241-6. https://doi.org/10.1055/s-2007-981613
  25. Kunikata T, Tatefuji T, Aga H, Iwaki K, Ikeda M, Kurimoto M. Indirubin inhibits inflammatory reactions in delayed-type hypersensitivity. Eur. J. Pharmacol.. 2000;410:93-100. https://doi.org/10.1016/S0014-2999(00)00879-7
  26. Hewett JA, Roth RA, Hepatic and extrahepatic pathobiology of bacterial lipopolysaccharides. Pharmacol. Rev. 1993;45:382-411.
  27. Watson WH, Zhao Y, Chawla RK. S-adenosylmethionine attenuates the lipopolysaccharide-induced expression of the gene for tumour necrosis factor alpha. Biochem. J. 1999;342:21-5. https://doi.org/10.1042/0264-6021:3420021
  28. Dinarello CA. Proinflammatory and antiinflammatory cytokines as mediators in the pathogenesis of septic shock. Chest. 1997;112:321S-9S. https://doi.org/10.1378/chest.112.6_Supplement.321S
  29. Vassalli P. The pathophysiology of tumor necrosis factors. Annu. Rev. Immunol. 1992;10:411-52. https://doi.org/10.1146/annurev.iy.10.040192.002211
  30. Goetz FW, Planas JV, MacKenzie S. Tumor necrosis factors. Dev. Comp. Immunol. 2004;28:487-97. https://doi.org/10.1016/j.dci.2003.09.008
  31. Casey LC, Balk RA, Bone RC. Plasma cytokine and endotoxin levels correlate with survival in patients with the sepsis syndrome. Ann. Intern. Med. 1993;119:771-8. https://doi.org/10.7326/0003-4819-119-8-199310150-00001
  32. Wogensen L, Jensen M, Svensson P, Worsaae H, Welinder B, Nerup J. Pancreatic beta-cell function and interleukin-1 beta in plasma during the acute phase response in patients with major burn injuries. Eur. J. Clin. Invest. 1993;23:311-9. https://doi.org/10.1111/j.1365-2362.1993.tb00780.x
  33. Shito M, Wakabayashi G, Ueda M, Shimazu M, Shirasugi N, Endo M, Mukai M, Kitajima M. Interleukin 1 receptor blockade reduces tumor necrosis factor production, tissue injury, and mortality after hepatic ischemia-reperfusion in the rat. Transplantation. 1997;63:143-8. https://doi.org/10.1097/00007890-199701150-00026
  34. Haeffner-Cavaillon N, Roussellier N, Ponzio O, Carreno M-P, Laude M, Carpentier A, Kazatchkine MD. Induction of interleukin-1 production in patients undergoing cardiopulmonary bypass. J. Thorac. Cardiovasc. Surg. 1989;98:1100-6.