Anti-inflammatory effects of Herba Artemisiae Capillaris as a consequence of the inhibition of NF-kappa B-dependent iNOS and pro-inflammatory cytokines production.

Nuclear Factor kappa B 억제를 통한 인진추출물의 inducible Nitric Oxide synthase 및 Cytokine 억제효과

  • Kim, Dae-Sung (College of Oriental Medicine, Daegu Haany University) ;
  • Park, Sook-Jahr (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) ;
  • Kim, Sang-Chan (College of Oriental Medicine, Daegu Haany University) ;
  • Byun, Sung-Hui (College of Oriental Medicine, Daegu Haany University)
  • 김대성 (대구한의대학교 한의과대학) ;
  • 박숙자 (대구한의대학교 한의과대학) ;
  • 조미정 (대구한의대학교 한의과대학) ;
  • 박상미 (대구한의대학교 한의과대학) ;
  • 김상찬 (대구한의대학교 한의과대학) ;
  • 변성희 (대구한의대학교 한의과대학)
  • Published : 2009.12.30

Abstract

Herba Artemisiae Capillaris is the dried bud of Artemisia capillaris Thunb, which has been used for expelling heat to loosen the bowels and normalizing gallbladder function to cure jaundice in traditional oriental medicines. In the present study, we evaluated the anti-inflammatory effects of the aqueous extracts of Herba Artemisiae Capillaris (HAC) in LPS-activated Raw 264.7 cells. Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h before adding HAC extract. Cell viability was determined by MTT assay, and the relative level of NO was measured with Griess reagent. TNF-$\alpha$, IL-$1{\beta}$, and IL-6 cytokines were detected by ELISA. During the entire experimental period, all three doses of HAC extract (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity. LPS-activated cells showed increased NO levels and iNOS expressions compared to control. However, these increases were dramatically attenuated by treatment with HAC extract. Moreover, the inhibitory effects of HAC extract occurred in a dose-dependent manner. In addition, HAC extract reduced the translocation of $NF{\kappa}B$ into nuclear. HAC reduced production of IL-$1{\beta}$ and IL-6 by LPS, although it had no effects on TNF-$\alpha$. These results demonstrate that liquiritigenin exerts anti-inflammatory effects, which results from the inhibition of $NF{\kappa}B$ activation in macrophages, thereby decreasing production of iNOS and proinflammatory cytokines. Taken together, these results indicate that the aqueous extracts of Herba Artemisiae Capillaris warrant further development as an anti-inflammatory agent for the treatment of gram-negative bacterial infections.

Keywords

References

  1. 신민교. 임상본초학. 서울:영림사. 1997:688-9
  2. 정보섭, 신민교. 도해향약대사전. 서울:영림사. 2003:1016-7.
  3. 김영철, 이장훈, 우홍정, 인진이 간성상세포의 섬 유화 억제에 미치는 영향에 대한 연구. 대한한 방내과학회지. 2005;26(4):853-63
  4. 신상만, 김영철, 이장훈, 우홍정. 인진이 TGF-$\beta$1 유도성 간섬유화에 미치는 영향. 대한한의학회지. 2001;22(3):141-55.
  5. 최인호, 김연희, 이동녕, 김형준. Candidaalbicans 에 대한 계지, 백두옹, 백선피, 백작약, 빈랑, 인진 의 항진균효과. 동의생리병리학회지. 2005;19(3):690-5.
  6. 함인혜, 정성웅, 이경진, 박규하, 최호영. 인진호 와 한인진이 Triton WR-1339로 유도된 고지혈 증 흰쥐에 미치는 영향. 대한본초학회지. 2005; 20(1):45-52.
  7. 박용진, 김영철, 이장훈, 우홍정. 인진 butanol 분획의 TLC추출성분이 Fas-mediated Apoptosis 에 미치는 영향. 대한한의학회지. 2002;23(2): 57-69.
  8. 이지현, 이장훈, 우홍정. 인진분획물이 HepG2 인체간암세포의 TGF$\beta$1-inducedapoptosis에 미치는 영향. 대한한의학회지. 2000;21(1):53-6
  9. 김이화, 김연희, 김연정, 장미현, 정주호, 김창주. 인진 약침액이 신경아세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향. 대한한의학회지. 2001;22(1):90-5.
  10. 강우성, 이장훈, 우홍정. 인진과 인진사령산가 감방이 간세포활성, 세포주기 및 DNA damage -inducedapoptosis에 미치는 영향. 대한한의학회지. 1999;20(1):91-105.
  11. 이채중, 김형환, 김종대, 김철호. 인진호의 열수 및 메탄올, 에테르, 부탄올 추출물이 CCI4유발에 의한 간독성 억제에 미치는 영향. 대한한방 내과학회지. 2000;21(1):100-7.
  12. 우홍정, 이장훈, 김영철. 인진과 갈근이 d-galactosamine, 급성 alcohol중독 및 CCl4 중독 백서의 간손상 에 미치는 영향. 대한한의학회지. 1997;18(1): 411-29.
  13. 이재동, 김재규. 인진수침이 Dimethylnitrosoamine 으로 유발된 흰쥐의 손상간에 미치는 영향. 경희대논문집. 1991;14:315-31
  14. 손갑호, 김성훈. 시호(柴胡), 인진(茵蔯)의 간 암세포에 대한 항암활성 및 항암제와의 상승 (相乘)작용. 대한한의학회지. 1995;16(2):414-32
  15. 박진상, 문석재, 문구, 원진희. 인진추출물이 마 우스 대식세포주인 RAW 264.7세포주의 iNOS 발현 및 superoxide형성에 미치는 영향, 한국전통의학회지, 1999;9(1):200-1
  16. LeeYS, Kim HS, Kim SK, Kim SD. IL-6 mRNA Expression in Mouse Peritoneal Macrophages and NIH3T3 Fibroblasts in Response to Candidaalbicans. J Microbiol Biotechnol. 2000; 10:8-15.
  17. HiguchiM, HigashiN, TakiH, Osawa T. Cytolytic mechanism of activated macrophages. Tumor necrosis factor and L-arginine-dependent mechanism acts as synergistically as the major cytolytic mechanism of activated macrophages. J Immunol. 1990;144:1425-31.
  18. McDaniel ML, Kwon G, Hill JR, Marshall CA and Corbett JA. Cytokines and nitric oxides in islet inflammation and diabetes. Proc.Soc.Exp.Biol.Med. 1996;211:24-32 https://doi.org/10.3181/00379727-211-43950D
  19. Corbett JA, and Mac Daniel ML. Intraislet release of interleukin-1 inhibits beta cell function by inducing beta cell expression of inducible nitric oxide syntheses. J.Exp.Med. 1995;181 :559-68. https://doi.org/10.1084/jem.181.2.559
  20. Cetkovic-Cvrlje M and Eizirik DL. TNF and IFN$\gamma$ potentiate the deleterious effects of IL-1$\beta$ on mouse pancreatic islets mainly via generation of nitric oxide. Cytokine.1994;6: 399-406. https://doi.org/10.1016/1043-4666(94)90064-7
  21. Kawamata H, Ochiai H, Mantani N, Terasawa K. Enhanced expression of inducible nitric oxide synthase by Juzen-taiho-to in LPS-activated RAW264.7 cells, a murine macrophage cell line. Am J Chin Med 2000;28:217-26. https://doi.org/10.1142/S0192415X0000026X
  22. 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 Meliaazedarach. Eur J Pharmacol. 2000 ;406:301-9. https://doi.org/10.1016/S0014-2999(00)00680-4
  23. 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
  24. 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 lipopolysaccharide-activated Raw 264.7 cells. Korean J. Oriental Physiology & Pathology. 2004;18(3):908-13.
  25. 장선일, 김형진, 황기명, 배현옥, 윤용갑, 정헌 택, 김윤철. 활성화된 설치류 RAW 264.7 대식 세포에서 당귀에탄올 추출물의 항염증 효과. 대한한의학방제학회지. 2002;10(2):189-197
  26. 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 cellline, RAW 264.7 cells. J Ethnopharmacol. 2001;76 :59-64. https://doi.org/10.1016/S0378-8741(01)00221-5
  27. 이영선, 한옥경, 신상우, 박종현, 권영규. 향부 자열수추출물의 Nitricoxide생성 및 iNOS 유전자 발현에 미치는 영향. 동의생리병리학회지. 2003;17(3):771-6.
  28. 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 Raw264.7 Cells. Evid Based Complement Alternat Med. 2007;4(3):327-33 https://doi.org/10.1093/ecam/nel093
  29. 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-kBa phosphorylation, C/EBP and AP-1 activation. British journal of pharmacology. 2003;139:11-20 https://doi.org/10.1038/sj.bjp.0705231
  30. ChenF, 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
  31. Delgado AV, McManus AT, ChambersJP. Production of tumor necrosis factor-alpha, interleukin1-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
  32. 본초학교수. 본초학. 서울:영림사. 1992:328-9
  33. 木村正康. 漢方藥理學(1). 東京:南山堂. 1997:311-2
  34. 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
  35. Kubesp. Inducible nitric oxide synthase; a little bit og good in allofus. Gut. 2000;47 :6-9. https://doi.org/10.1136/gut.47.1.6
  36. Chiou WF, Chou CJ, Chen CF. Camptothecin suppresses nitric oxide biosynthesis in RAW 264.7 macrophages. Life Sci. 2001;69:625-35. https://doi.org/10.1016/S0024-3205(01)01154-7
  37. Seo WG, Pae HO, Oh GS, Kim NY, Kwon TO, Shin MK, Chai KY, Chung HT. The aqueous extract of Rhodiola sachalinensis root enhances the expression of inducible nitric oxide synthase gene in RAW264.7 macrophages. J Ethnopharmacol. 2001;76:119-23. https://doi.org/10.1016/S0378-8741(01)00220-3
  38. 박혜준, 윤화정, 윤정원, 윤소원, 고우신. 오미 소독음이 마우스 복강 대식세포에서 NO의 분 비과정에 미치는 영향. 동의생리병리학회지. 2002;16(2):921-7.
  39. Casey LC, Balk RA, Bone RC. Plasma cytokine and endotoxin levels correlate with survival in patients with the sepsis syndrome. AnnIntern Med. 1993;119:771-8.
  40. Wogensen L, Jensen M, Svensson P, Worsaae H, Welinder B, Nerup J. Pancreatic beta-cell function and interleukin-1$\beta$ inplasmaduring the acute phase response in patients with majorburninjuries. Eur J ClinInvest. 1993 ;23:311-9. https://doi.org/10.1111/j.1365-2362.1993.tb00780.x
  41. Shito M, Wakabayashi G, Ueda M, Shimazu M, Shirasugi N, Endo M, etal. Interleukin 1 receptor blockade reduces tumor necrosis factor production, tissue injury, and mortality after hepatic ischemia-reperfusion in therat. Transplantation. 1997;63:143-8. https://doi.org/10.1097/00007890-199701150-00026
  42. Takabayashi T, Shimizu S, Clark BD, Beinborn M, Burke JF, Gelfand JA. Interleukin-1 upregulates anaphylatoxin receptorson mononuclear cells. Surgery. 2004;135(5):544-54. https://doi.org/10.1016/j.surg.2003.09.010