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

Effects of Polygoni Cuspidati Radix on the $H_2O_2$-treated LLC-$PK_1$ Cell's Redox Status and NF-${\kappa}B$ Signaling

호장근(虎杖根)이 $H_2O_2$에 노출된 LLC-$PK_1$ 세포의 Redox Status 및 NF-${\kappa}B$ Signaling에 미치는 영향

  • Kim, Sol-Ri (Department of Internal Medicine, College of Oriental Medicine, Dongguk University) ;
  • Jeong, Ji-Cheon (Department of Internal Medicine, College of Oriental Medicine, Dongguk University)
  • 김솔리 (동국대학교 한의과대학 내과학교실) ;
  • 정지천 (동국대학교 한의과대학 내과학교실)
  • Received : 2012.07.05
  • Accepted : 2012.08.20
  • Published : 2012.08.25
Download PDF
⟨ Previous Next ⟩

Abstract

This study was designed to identify the effects of Polygoni cuspidati Radix(PCR) on the generation of superoxide anion radicals (${\cdot}O_2{^-}$), nitric oxide (NO), peroxynitrite ($ONOO^-$) in the renal epithelial cells of mouse(LLC-$PK_1$). The effects of PCR on the expression of inflammation-related proteins, IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1, were examined by western blotting. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 2',7'-dichloro dihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) were used. Protein expression levels of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1 were assayed by western blot. PCR reduced $H_2O_2$-induced cell death dose-dependently. It inhibited the generation of ${\cdot}O_2{^-}$, NO, $ONOO^-$ and $PGE^2$ in the $H_2O_2$-treated LLC-PK1 cells in vitro. PCR inhibited the espression of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest that PCR is an effective NO, ${\cdot}O_2{^-}$, $ONOO^-$ scavenger, and this substance recommended to be applied in treatment for the inflammatory process and inflammation-related disease.

Keywords

References

  1. Yu, B.P. Aging and oxidative stress: modulation by dietary restriction. Free Radic Biol Med. 21: 651-668, 1996. https://doi.org/10.1016/0891-5849(96)00162-1
  2. Helmer, K.S., Chang, L., Cui, Y., and Mercer, D.W. Induction of NF-${\kappa}B$, $I{\kappa}B-{\alpha}$, and iNOS in rat gastric mucosa during endotoxemia. J Surg Res. 104(1):46-52, 2002. https://doi.org/10.1006/jsre.2002.6404
  3. Carr, A.C. Oxidation of LDL by myeloperoxidase and reactive nitrogen species: reaction pathways and antioxidant protection. Arterioscler Thromb Vasc Biol. 20: 1716-1723, 2000. https://doi.org/10.1161/01.ATV.20.7.1716
  4. Haenen, G.R., Paquay, J.B., Korthouwer, R.E., and Bast, A. Peroxynitrite scavenging by flavonoids. Biochem Biophy Res Commu. 236(3):591-593, 1997. https://doi.org/10.1006/bbrc.1997.7016
  5. Kim, H.J., Kim, K.W., Yu, B.P., and Chung, H.Y. The effct of age on cyclooxygenase-2 gene expression: NF-${\kappa}B$ activation and $I{\kappa}B{\alpha}$ degradation. Free Radic Biol Med. 28(5):683-692, 2000. https://doi.org/10.1016/S0891-5849(99)00274-9
  6. Baeuerle, P.A., and Henkei, T. Function and activation of NF-${\kappa}B$ in the immune system. Annu Rev Immunol. 12: 141-179, 1994. https://doi.org/10.1146/annurev.iy.12.040194.001041
  7. Nathan, C.F., and Hibbs, J.B. Roles of nitric oxide synthesis in macrophage antimicrobial activity. Curr Opin Immunol. 1: 65-70, 1991.
  8. Kawamori, T., Rao, C.V., Seibert, K., and Reddy, B.S. Chemopreventive activity of elecoxib, a pecific cyclooxygenase-2 Inhibitor, against colon carcinogenesis. Cancer Res. 58: 409-412, 1998.
  9. Lawrence, T., Gilroy, D.W., Colville-Nash, P.R., and Willoughby, D.A. Possible new role for NF-${\kappa}B$ in the resolution of inflammation. Nat Med. 7(12):1291-1297, 2001. https://doi.org/10.1038/nm1201-1291
  10. Korhonen, P., Helenius, M., and Salminen, A. Age-related chages in the reguration of transcription factor NF-kappa B in rat brain. Neurosci Lett. 225: 61-64, 1997. https://doi.org/10.1016/S0304-3940(97)00190-0
  11. 全國韓醫科大學共同敎材編纂委員會編著. 本草學. 서울, 永林社, pp 462-463, 2005.
  12. 정운석, 이주일, 서운교. 백일해 유발균 Bordetella pertussis 에 대한 虎杖根의 생육억제 효과. 대한한방내과학회지 26(3):543-550, 2005.
  13. 이태규, 김종화, 소준노. 虎杖根으로부터 분리된 emodin의 혈관신생 억제 활성. 한국농화학회지 46(1):50-54, 2003.
  14. 김동조, 김호현, 고흥. 虎杖根물추출물의 내피세포 의존성 혈관이완효과. 동의생리병리학회지 22(1):131-136, 2008.
  15. 최변탁, 김연희, 이동녕, 김형준. 虎杖根이 子宮頸部癌細胞의 成長抑制및 細胞枯死에 미치는 영향. 대한한방부인과학회지 19(1):1-13, 2006.
  16. 김지연, 양기숙. 虎杖根의 사염화탄소로 유도된 지질과산화 저해활성. 약학회지 43(5):572-576, 1999.
  17. 이인선, 임효권, 이승욱. 虎杖根에 의한 Helicobacter pylori의 생육 저해. 한국식품과학회지 35(6):1182-1187, 2003.
  18. Tada, H., Shiho, O., Kuroshima, K., Koyama, M., and Tsukamoto, K. An improved colorimetric assay for interleukin 2. J Immunol Methods. 93(2):157-165, 1986. https://doi.org/10.1016/0022-1759(86)90183-3
  19. Cathcart, R., Schwiers, E., and Ames, B.N. Detection of picomole levels of hydroperoxides using a fluorescent dichlorofluorescein fluorescent assay. Anal Biochem. 134: 111-116, 1983. https://doi.org/10.1016/0003-2697(83)90270-1
  20. Nagata, N., Momose, K., and Ishida, Y. Inhibitory effects of catecholamines and anti-oxidants on the fluorescence reaction of 4,5-diaminofluorescein, DAF-2, a novel indicator of nitric oxide. J Biochem Tokyo. 125: 658-661, 1999. https://doi.org/10.1093/oxfordjournals.jbchem.a022333
  21. Kooy, N.W., Royall, J.A., Ischiropoulos, H., and Beckman, J.S. Peroxynitrite-mediated oxidated oxidation of dihydrorhodamine 123. Free Radic Res Commun. 16: 149-1456, 1994.
  22. Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. Protein measurement with the Folin phenol reagent. J Biol Chem. 193(1):265-275, 1951.
  23. 陶弘景. 名醫別錄. 북경, 人民衛生出版社, pp 228-229, 1986.
  24. 許浚. 東醫寶鑑. 서울, 법인문화사, p 1962, 1999.
  25. Qian, G., Leung, S., Lu, G., and Leung, K. Differentiation of Rhizoma Et Radix Poligoni Cuspidati from Closely Related Herbs by HPLC Fingerprinting. Chem Pharm Bull. 54: 1179-1186, 2006. https://doi.org/10.1248/cpb.54.1179
  26. Sato, M., Maulik, G., Bagchi, D., and Das, D.K. Myocardial protection by protykin, a novel extract of trans-resveratrol and emodin. Free Radic Res. 32: 135-144, 2000. https://doi.org/10.1080/10715760000300141
  27. Jayasuriya, H., Koonchanok, N.M., Geahlen, R.L., MacLaughlin, J.L., and Chang, C.J. Emodin, a protein kinase inhibitor from Polygonum cuspidatum. J. Nat. Prod. pp 55, 696-698, 1992. https://doi.org/10.1021/np50083a026
  28. Kumar, A., Dhawan, S., and Aggarwal, B.B. Emodin (3-methyl-1,6,8-trihydroxyanthraquinone) inhibits TNF-induced NF-${\kappa}B$ activation, IkB degradation, and expression of cell surface adhesion proteins in human vascular endothelial cells. Oncogene pp 17, 913-918, 1998. https://doi.org/10.1038/sj.onc.1201998
  29. 최병진. 한약의 약리. 서울, 고려의학, pp 117-118, 339, 2000.
  30. Mates, J.M., Perez-Gomez, C., and Nunez de Castro I. Antioxidant enzymes and human disease. Clin. Biochem. 32(8):595-603, 1999. https://doi.org/10.1016/S0009-9120(99)00075-2
  31. Shingu, M., Oribe, M., Todoroki, T., Tatsukawa, K., Tomo-Oka, K., Yasuda, M., and Nobunaga, M. Serum factors from patients with systemic lupus erythematosus enhancing superoxide generation by normal neutrophils. J. Invest. Dermatol. 81(3):212-215, 1983. https://doi.org/10.1111/1523-1747.ep12517989
  32. Pannala, A.S., Rice-Evans, C.A., Halliwell, B., and Singh, S. Inhibition of peroxynitrite- mediated tyrosine nitration by catechin polyphenols. Biochem Biophy Res Commu. 232: 164-170, 1997. https://doi.org/10.1006/bbrc.1997.6254
  33. Shimokawa, H., Zanchetti, A., and Mancia, G. Handbook of Hypertension. Elvsevier Science. Amsterdam. pp 1007-1068, 1997.
  34. Chung, H.Y., Kim, H.j., and Kim, J.W. The inflammation hypothesis of aging: Molecular modulation by calorie restriction. Ann N Y Acad Sci. 928: 327-335, 2001.
  35. Crow, J.P., Beckman, J.S., and McCord, J.M. Sensitivity of the essential zinc-thiolate moiety of yeast alcohol dehydrogenase to hypochlorite and peroxynitrite. Biochemistry. 34(11):3533-3552, 1995.
  36. 정해영, 성도유, 김애라, 최혜리, 김현진, 최재수. ONOO-의 생성, 독성, 제거 및 노화와 관련성. 한국노화학회지 10(1):46-59, 2000.
  37. Halushka, P.V., Mais, D.E., Mayeux, P.R., and Morinelli, T.A. Thromboxane, prostaglandin and leukotriene receptors. Annu Rev Pharmacol Toxicol. 29: 213-239, 1989. https://doi.org/10.1146/annurev.pa.29.040189.001241
  38. Masferrer, J., Zweofel, B.S., Manning, P.T., Hauser, S.D., Leahy, K.M., Smith, W.G., Isacson, P.C., and Seibert, K. Selective inhibition of inducible cyclooxygenase 2 in vivo is anti-inflammatory and nouulcerogenic. Proc. Natl. Acad. U.S.A. 91: 3228-3232, 1994. https://doi.org/10.1073/pnas.91.8.3228
  39. Seibert, K., Zhang, Y., Leahy, K., Hauser, S., Masferrer, J., Perkins, W., Lee, L., and Ksakson, P. Pharmacoogical and biochemical demonstration and pain. Proc. Natl. Acad. Sci. U.S.A. 91: 12013-12017, 1994. https://doi.org/10.1073/pnas.91.25.12013
  40. Groszmann, R.J. Hyperdynamic state in chronic liver disease. J Hepatol. 224(1):61-64, 1993.
  41. Chung, H.Y., Kim, H.J., Kim, K.W., Choi, J.S., and Yu, B.P. Molecular inflammation hypothesis of aging based on the anti-aging mechanism of calorie restriction. Microsc Res Tech. 59(4):264-272, 2002. https://doi.org/10.1002/jemt.10203
  42. 김선영 외 14명 역. 면역생물학 6판. 서울, 라이프사이언스, p 78, 237, 2005.
  43. Alagappan, V.K., McKay, S., Widyastuti, A., Garrelds, I.M., Bogers, A.J., and Hoogsteden, H.C. Proinflammatory cytokines upregulate mRNA expression and secretion of vascular endothelial growth factor in cultured human airway smooth muscle cells. Cell Biochem Biophys. 43(1):119-129, 2005. https://doi.org/10.1385/CBB:43:1:119
  44. Pannala, A.S., Rice-Evans, C.A., Halliwell, B., and Sin, S. Inhibition of peroxynitrite-mediated tyrosine nitration by catechin polyphenoles. Biochem Biophy Res Commu. 232: 164-170, 1997. https://doi.org/10.1006/bbrc.1997.6254