Journal of Physiology & Pathology in Korean Medicine (동의생리병리학회지)

The Physiological Society of Korean Medicine and The Society of Pathology in Korean Medicine (대한동의생리학회·한의병리학회)
권호 표지

Inhibitory Effects of Hoechunyanggyeok-san on Inflammation in Vivo and in Vitro

회춘양격산 물추출물의 항염증효과

  • 이태호 (대구한의대학교 한의과대학) ;
  • 조미정 (대구한의대학교 한방신약개발팀(BK21 Team)) ;
  • 박숙자 (대구한의대학교 한방신약개발팀(BK21 Team)) ;
  • 이종록 (대구한의대학교 한의과대학) ;
  • 백영두 (대구한의대학교 한의과대학) ;
  • 김종열 (한국한의학연구원) ;
  • 권영규 (부산대학교 한의전문대학원) ;
  • 김상찬 (대구한의대학교 한방신약개발팀(BK21 Team))
  • Published : 2009.12.25
Download PDF
⟨ Previous Next ⟩

Abstract

Hoechunyanggyeok-san (HYS) is a traditional oriental herbal medicine widely used for treating inflammatory disorders. Although there are numerous clinical results of HYS reported in the literature of oriental hebal medicine, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of HYS extract (HYSE) in vivo and in vitro. To determine the cytotoxic concentration of HYSE, cell viability was tested by MTT assay. All four doses of HYSE (0.01, 0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. In order to measure NO levels in culture medium, the cells were treated with $1\;{\mu}g/ml$ of LPS 1h before adding HYSE for 24 h and then culture medium were reacted with Griess reagent. Increased NO production and iNOS expression were detected in LPS-activated cells compared to control. However, these increases were dose-dependently attenuated by treatment with HYSE. LPS plays a key role in leading to the massive production of pro-inflammatory cytokines such as TNF-$\alpha$, IL-$1{\beta}$ and IL-6 in macrophages. Thus, we next determined the levels of these cytokines. HYSE reduced the elevated production of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 by LPS. Moreover, the effects of HYSE were in a dose-dependent manner. In vivo, histopathological study, HYSE effectively inheefed the increases of hind paw skin thicknesses and inflammatory cell infiltrations induced by carrageenan treatment. It, therefore, considered that HYSE will be favorably inheefed the acute edematous inanner. In s. These findings showed that HYSE could have anti-inflammatory effects through the reduction of NO and inflammatory cytokines in macrophage. Furthermore, the reduction of carrageenan-induced paw oedema by HYSE helps to understand its actions on inflammatory conditions.

Keywords

References

  1. 허 준. 동의보감. 남산당, 서울, p 252, 420, 1987
  2. 김경준, 김중호, 채병윤. 회춘양격산이 항알레르기 및 소염 진통 해열효과에 미치는 영향. 대한외관과학회지 7: 1-13, 1994
  3. 강승원, 노석선. 回春凉膈散과 龍石散이 抗炎作用에 미치는 影響. 대한외관과학회지 12(1):47-78, 1999
  4. 한유진, 이용태, 장경전. Carrageenan 유발염증에 대한 15Hz 전침의 효과에 대한 연구. 대한침구학회지 20(3):166-176, 2003
  5. 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. 144: 1425-1431, 1990
  6. 국윤범. 황련해독탕이 자발적 고혈압 백서의 혈압 및 신장 기능에 미치는 영향. 대한한의방제학회지 10(1):113-129, 2002
  7. 黃道淵 原著. 南山堂編輯局 飜譯. 對譯證脈方藥合編. 南山堂. 서울, pp 229-230, 1985
  8. Kim, H.D., Cho, H.R., Moon, S.B., Shin, H.D., Yang, K.J., Park, B.R., Jang, H.J., Lim, L.S., Lee, H.S., Ku, S.K. Effect of exopolymers from Aureobasidum pullulans on formalin-induced chronic paw inflammation in mice. J Microbiol Biotechnol. 16: 1954-1960, 2006
  9. Lee, A.K., Sung, S.H., Kim, Y.C., Kim, S.G. Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-a and COX-2 expression by sauchinone effects on I-κBa phosphorylation, C/EBP and AP-1 activation. British journal of pharmacology 139: 11-20, 2003 https://doi.org/10.1038/sj.bjp.0705231
  10. Delgado, A.V., McManus, A.T., Chambers, J.P. Production of tumor necrosis factor-alpha, interleukin 1-beta, interleukin 2, and interleukin 6 by rat leukocyte subpopulations after exposure to substance P. Neuropeptides. 37(6):355-361, 2003 https://doi.org/10.1016/j.npep.2003.09.005
  11. Jirik, F.R., Podor, T.J., Hirano, T., Kishimoto, T., Loskutoff, D.J., Carson, D.A., Lotz, M. Bacterial lipopolysaccharide and inflammatory mediators augment IL-6 secretion by human endothelial cells. J Immunol. 142(1):144-147, 1989
  12. Mori, M. Regulation of nitric oxide synthesis and apoptosis by arginase and arginine recycling. J Nutr. 137: 1616S-1620S, 2007 https://doi.org/10.1093/jn/137.6.1616S
  13. McDaniel, M.L., Kwon, G., Hill, J.R., Marshall, C.A. and Corbett, J.A. Cytokines and nitric oxides in islet inflammation and diabetes. Proc. Soc. Exp. Biol. Med. 211: 24-32, 1996 https://doi.org/10.3181/00379727-211-43950D
  14. Corbett, J.A. and Mac, Daniel, M.L. Intraislet release of interleukin-1 inhibits beta cell function by inducing beta cell expression of inducible nitric oxide syntheses. J. Exp. Med. 181: 559-568, 1995 https://doi.org/10.1084/jem.181.2.559
  15. Cetkovic Cvrlje, M. and Eizirik, D.L. TNF and IFNγ potentiate the deleterious effects of IL-1$\beta$ on mouse pancreatic islets mainly via generation of nitric oxide. Cytokine. 6: 399-406, 1994 https://doi.org/10.1016/1043-4666(94)90064-7
  16. 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. 28: 217-226, 2000 https://doi.org/10.1142/S0192415X0000026X
  17. Lee, B.G., Kim, S.H., Zee, O.P., Lee, K.R., Lee, H.Y., Han, J.W., Lee, H.W. Suppression of inducible nitric oxide synthase expression in RAW 264.7 macrophages by two-carboline alkaloids extracted from Melia azedarach. Eur J Pharmacol. 406: 301-309, 2000 https://doi.org/10.1016/S0014-2999(00)00680-4
  18. 장선일, 김형진, 황기명, 배현옥, 윤용갑, 정헌택, 김윤철. 활성화된 설치류 RAW 264.7 대식세포에서 당귀에탄올 추출물의 항염증 효과. 대한한의학방제학회지 10(2):189-197, 2002
  19. Seo, W.G., Pae, H.O., Oh, G.S., K.Y. Chai, Kwon, T.O., Y.G. Yun, N.Y. Kim, H.T. 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. 76: 59-64, 2001 https://doi.org/10.1016/S0378-8741(01)00221-5
  20. 이영선, 한옥경, 신상우, 박종현, 권영규. 향부자 열수추출물의 Nitric oxide 생성 및 iNOS 유전자 발현에 미치는 영향. 동의생리병리학회지 17(3):771-776, 2003
  21. Chun, S.C., Jee, S.Y., Lee, S.G., Park, S.J., Lee, J.R., Kim, S.C. Anti-Inflammatory Activity of the Methanol Extract of Moutan Cortex in LPS-Activated Raw264.7 Cells. Evid Based Complement Alternat Med. 4(3):327-333, 2007 https://doi.org/10.1093/ecam/nel093
  22. Sharifi, A.M., Mousavi, S.H., Bakhshayesh, M., Tehrani, F.K., Mahmoudian, M., Oryan, S. Study of correlation between lead-induced cytotoxicity and nitric oxide production in PC12 cells. Toxicol Lett. 160: 43-48, 2005 https://doi.org/10.1016/j.toxlet.2005.06.008
  23. 박은영, 박숙자, 이종록, 지선영, 변성희, 김상찬. PC-12 cell에서 감초성분의 Liquiritigenin이 납에 의해 유도된 세포독성과 nitric oxide production에 미치는 영향, 본초학회지 22(2):17-24, 2007
  24. Palmer, R.M., Ashton, D.S., Moncada, S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature. 333: 664-666, 1988 https://doi.org/10.1038/333664a0
  25. Lee, J.H., Choi, Y.H., Choi, B.T. The anti-inflammatory effects of 2 Hz electroacupuncture with different intensities on acute carrageenan-induced inflammation in the rat paw. Int J Mol Med. 16: 99-102, 2005
  26. Gupta, M., Mazumder, U.K., Gomathi, P., Selvan, V.T. Antiinflammatory evaluation of leaves of Plumeria acuminata. BMC Complement Altern Med. 6: 36, 2006 https://doi.org/10.1186/1472-6882-6-36
  27. Rao, C.V., Verma, A.R., Gupta, P.K., Vijayakumar, M. Anti-inflammatory and anti-nociceptive activities of Fumaria indica whole plant extract in experimental animals. Acta Pharm. 57: 491-498, 2007 https://doi.org/10.2478/v10007-007-0039-z
  28. Holt, S., Comelli, F., Costa, B., Fowler, C.J. Inhibitors of fatty acid amide hydrolase reduce carrageenan-induced hind paw inflammation in pentobarbital-treated mice: comparison with indomethacin and possible involvement of cannabinoid receptors. Br J Pharmacol. 146: 467-476, 2005 https://doi.org/10.1038/sj.bjp.0706348
  29. Liu, J., Zhang, W., Zhou, L., Wang, X., Lian, Q. Anti-inflammatory effect and mechanism of osthole in rats. Zhong Yao Cai. 28: 1002-1006, 2005
  30. Beloeil, H., Ababneh, Z., Chung, R., Zurakowski, D., Mulkern, R.V., Berde, C.B. Effects of bupivacaine and tetrodotoxin on carrageenan-induced hind paw inflammation in rats (Part 1): hyperalgesia, edema, and systemic cytokines. Anesthesiology. 105: 128-138, 2006 https://doi.org/10.1097/00000542-200607000-00022
  31. Kim, Y.W., Zhao, R.J., Park, S.J., Lee, J.R., Cho, I.J., Yang, C.H., Kim, S.G., Kim, S.C. Anti-inflammatory effects of liquiritigenin as a consequence of the inhibition of NF-kappaB-dependent iNOS and proinflammatory cytokines production. Br J Pharmacol. 154(1):165-173, 2008 https://doi.org/10.1038/bjp.2008.79