DOI QR코드

DOI QR Code

LPS로 활성화된 대식세포에서 황련해독탕(黃連解毒湯) 물추출물의 염증매개물질 억제효과

Anti-inflammatory Effects of the Aqueous Extract of Hwangnyeonhaedok-tang in LPS-activated Macrophage Cells

  • 김대희 (대구한의대학교 한의과대학) ;
  • 박숙자 (대구한의대학교 한방신약개발팀(BK21 Team)) ;
  • 정지윤 (대구한의대학교 한방신약개발팀(BK21 Team)) ;
  • 김상찬 (대구한의대학교 한방신약개발팀(BK21 Team)) ;
  • 변성희 (대구한의대학교 한의과대학)
  • 발행 : 2009.12.30

초록

Objectives : Hwangnyeonhaedok-tang (Huanglian Jiedu Tang; HHT) has been widely used for purging' 'fire' and lessening virulence of any pathogenic organism. However it has been rarely conducted to evaluate the immuno-biological activity. In this study, we evaluated anti-inflammatory effects of HHT in LPS-activated Raw264.7 cells. Methods : Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h prior to the addition of HHT. Cell viability was measured by MTT assay. The production of NO was determined by reacting cultured medium with Griess reagent. PGE2 and proinflammatory cytokines were detected by ELISA. Expression of iNOS, COX-2, $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ were analyzed by immunoblot analysis. Results : All three doses of HHT (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. The levels of NO and PGE2 were dramatically augmented by LPS compared to control. However, HHT extract dose-dependently reduced these increases. Expression of iNOS and COX-2 protein were also decreased by treatment with HHT extract. Furthermore, HHT extract significantly reduced the nuclear translocation of NF-${\kappa}B$ which is critical in regulating inflammation through transcription of iNOS and COX-2. In addition, HHT extract reduced the elevated production of inflammatory cytokines including TNF-$\alpha$, IL-$1{\beta}$ and IL-6. Conclusions : The results in this study demonstrate that HHT extract exerts anti-inflammatory activities through the inhibition of NO, PGE2 and proinflammatory cytokines production via the suppression of NF-${\kappa}B$.

키워드

참고문헌

  1. 葛洪. 肘後備急方. 上海 : 人民衛生出版社. 1996 : 34-7.
  2. 방제학교수. 방제학. 서울 : 영림사. 1999 : 180
  3. Kawashima K, Haruo N, Kogure K. Effect of Oren-gedoku-to on cerebral vascular accident. Pharma Medica. 1988 ; 6 : 33-6.
  4. Ohta Y, Sasaki E, Nishida K, Hayashi T, Nagata M, Ishiguro I. Preventive effect of Oren-gedokuto( Huang-lian-jie-do-tang)extract lipid peroxidation in rat liver microsomes. Biol Pharm Bull. 2001 ; 24(10) : 1165-70. https://doi.org/10.1248/bpb.24.1165
  5. Hayashi T, Ohta Y, Inagaki S, Harada N. Inhibitory action of Oren-Gediku-to extract on enzymatic lipid peroxidation in rat liver microsomes. Biol Pharm Bull. 2001 ; 24(10) : 1165-70. https://doi.org/10.1248/bpb.24.1165
  6. Mizukawa H, Yoshida K, Honmura A, Uchiyama Y, Kaku H, Nakajima S, Haruki E. The effect of oregedokuto on experimentally-inflamed rats. American Journal of Chinese Medicine. 1993 ; 21 : 71-8. https://doi.org/10.1142/S0192415X93000091
  7. 이병철, 임강현, 김영옥, 김선여, 안덕균, 박호균, 김호철. 황금(黃芩)의 4-VO로 유발한 흰쥐뇌허혈에 대한 신경방어효과. 대한본초학회지. 1999 ; 14 : 89-96.
  8. 김호철. 한약리학. 서울 : 집문당. 2001 : 125-40.
  9. 김보애, 김미소, 강보미, 변선희, 박일향, 박지하, 정지욱, 안은미, 정현아, 장정희, 배원, 이하영, 최필녀,박찬익. 황련해독탕이 NC/Nga Mice에서 유발된 아토피 피부염에 미치는 영향. 대한본초학회지. 2008 ; 23(2) : 59-65.
  10. 김광호, 김성수. 황련해독탕과 온성어혈방의 수침 및 경구투여가 진통 소염 해열작용에 미치는 영향. 대한한의학회지. 1994 ; 15(1) : 9-25.
  11. 박수현, 권용욱, 이태희. 황련해독탕과 건강부자탕이 LPS 유도에 의한 마우스 혈중 IL-6와 TNF-$\alpha$변화 에 미치는 영향. 대한한의방제학회지. 2007 ; 15(1) :185-97.
  12. Lee YS, Kim HS, Kim SK, Kim SD. IL-6 mRNA Expression in Mouse Peritoneal Macrophages and NIH3T3 Fibroblasts in Response to Candida albicans. J Microbiol Biotechnol. 2000 ; 10 : 8-15.
  13. Higuchi M, Higashi N, Taki H, Osawa T. Cytolytic mechanism of activated macrophages. Tumor necrosis factor and L-arginine-dependent mechanism acts as synergistically as the major cytolytic mechanism of activated macrophages. J Immunol. 1990 ; 144 : 1425-31.
  14. McDaniel ML, Kwon G, Hill JR, Marshall CA and Corbett JA. Cytokines and nitric oxides in islet inflammation and diabetes. Proc Soc Exp Biol Med. 1996 ; 211 : 24-32.
  15. Kawamata H, Ochiai H, Mantani N, Terasawa K. Enhanced expression of inducible nitric oxide synthase by Juzen-taiho-to in LPS-activated Raw 264.7 cells, a murine macrophage cell line. Am J Chin Med. 2000 ; 28 : 217-26. https://doi.org/10.1142/S0192415X0000026X
  16. Lee BG, Kim SH, Zee OP, Lee KR, Lee HY, Han JW, Lee HW. Suppression of inducible nitric oxide synthase expression in RAW 264.7 macrophages by two-carboline alkaloids extracted from Melia azedarach. Eur J Pharmacol. 2000 ; 406 : 301-9. https://doi.org/10.1016/S0014-2999(00)00680-4
  17. Seo WG, Pae HO, Oh GS, Chai KY, Yun YG, Kwon TO, Chung HT. Inhibitory effect of ethyl acetate fraction from Cudrania tricuspidata on the expression of nitric oxide synthase gene in RAW 264.7 macrophages stimulated with interferon-and lipopolysaccharide. Gen Pharmacol. 2000 ; 35 : 21-8. https://doi.org/10.1016/S0306-3623(01)00086-6
  18. Yoon TG, Byun BH, Kwon TK, Suh SI, Byun SH, Kwon YK, Kim SC. Inhibitory effect of Farfarae Flos water extract on COX-2, iNOS expression and nitric oxide production in lipopolysaccharideactivated Raw 264.7 cells. Korean J Oriental Physiology & Pathology. 2004 ; 18(3) : 908-13.
  19. 장선일, 김형진, 황기명, 배현옥, 윤용갑, 정헌택, 김윤철. 활성화된 설치류 RAW 264.7 대식세포에서 당귀에탄올 추출물의 항염증 효과. 대한한의학방제학회지. 2002 ; 10(2) : 189-97.
  20. Seo WG, Pae HO, Oh GS, KY Chai, Kwon TO, YG Yun, NY Kim, HT Chung. Inhibitory effects of methanol extract of Cyperus rotundus rhizomes on nitric oxide and superoxide production by murine macrophage cell line, RAW 264.7 cells. J Ethnopharmacol. 2001 ; 76 : 59-64. https://doi.org/10.1016/S0378-8741(01)00221-5
  21. Chun SC, Jee SY, Lee SG, Park SJ, Lee JR, Kim SC. Anti-Inflammatory Activity of the Methanol Extract of Moutan Cortex in LPS-Activated Raw 264.7 Cells. Evid Based Complement Alternat Med. 2007 ; 4(3) : 327-33. https://doi.org/10.1093/ecam/nel093
  22. 국윤범. 황련해독탕이 자발적 고혈압 백서의 혈압 및 신장 기능에 미치는 영향. 대한한의방제학회지.2002 ; 10(1) : 113-29.
  23. Nathan, C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992 ; 6 : 3051-64. https://doi.org/10.1096/fasebj.6.12.1381691
  24. 손갑호, 김성훈. 시호(柴胡), 인진(茵蔯)의 간암세포에 대한 항암활성 및 항암제와의 상승(相乘)작용. 대한한의학회지. 1995 ; 16(2) : 414-32.
  25. 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. British Journal of Pharmacology. 2003 ; 139 : 11-20. https://doi.org/10.1038/sj.bjp.0705231
  26. 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(6) : 355-61. https://doi.org/10.1016/j.npep.2003.09.005
  27. Jirik FR, Podor TJ, Hirano T, Kishimoto T, Loskutoff DJ, Carson DA, Lotz M. Bacterial lipopolysaccharide and inflammatory mediators augment IL-6 secretion by human endothelial cells. J Immunol. 1989 ; 142(1) : 144-7.
  28. Chen F, Castranova V, Shi X. New insights into the role of nuclear factor-kappaB in cell growth regulation. Am J Pathol. 2001 ; 159(2) : 387-97. https://doi.org/10.1016/S0002-9440(10)61708-7
  29. Mori M. Regulation of nitric oxide synthesis and apoptosis by arginase and arginine recycling. J Nutr. 2007 ; 137 : 1616S-20S. https://doi.org/10.1093/jn/137.6.1616S
  30. Sharifi AM, Mousavi SH, Bakhshayesh M, Tehrani FK, Mahmoudian M, Oryan S. Study of correlation between lead-induced cytotoxicity and nitric oxide production in PC12 cells. Toxicol Lett. 2005 ; 160 : 43-8. https://doi.org/10.1016/j.toxlet.2005.06.008
  31. 박은영, 박숙자, 이종록, 지선영, 변성희, 김상찬. PC-12 cell에서 감초성분의 Liquiritigenin이 납에 의해 유도된 세포독성과 nitric oxide production에 미치는 영향. 본초학회지. 2007 ; 22(2) : 17-24.
  32. Palmer RM, Ashton DS, Moncada S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature. 1988 ; 333 : 664-6. https://doi.org/10.1038/333664a0
  33. 국윤범. 황련해독탕과 건강부자탕이 LPS 유도에 의 한 마우스 혈중 IL-6와 TNF-$\alpha$ 변화에 미치는 영향. 대한한의학방제학회지. 2007 ; 15(1) : 185-97.
  34. 양창헌. 흔히 접하는 비스테로이드성 항염증약물 (NSAIDs)과 위장관질환. 동국의학. 2003 ; 10(2) : 190-9.
  35. Taylor PC, Williams RO, Feldmann M. Tumour necrosis factor alpha as a therapeutic target for imunne-mediated inflammatory disease. Curr Opin Biotechnol. 2004 ; 15 : 557-3. https://doi.org/10.1016/j.copbio.2004.09.005
  36. 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
  37. 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
  38. 이순희, 임병우, 조여원. DSS로 유도된 염증성 장질환 동물 모델에서 황금 열수 추출물이 면역 조절기능에 미치는 영향. 한국영양학회지. 2004 ; 37(6) :431-9.
  39. 박용기, 이경열. 황련 클로로포름 분획물의 뇌신경소교세포로부터 염증매개물질 생성억제 효능 연구. 대한본초학회지. 2007 ; 22(4) : 109-6.