Effects of Vespae Nidus on Peroxynitrite Production and Protein Expression of Proinflammatory Mediators

노봉방(露蜂房)이 t-butylhydroxyperoxide에 의한 Peroxynitrite 생성과 염증성 단백질 발현에 미치는 영향

  • Jang, Jae-Shik (Department of Internal Medicine, College of Oriental Medicine, Dongguk University) ;
  • Jeong, Ji-Cheon (Department of Internal Medicine, College of Oriental Medicine, Dongguk University) ;
  • Shin, Hyeon-Cheol (Department of Internal Medicine, College of Oriental Medicine, Daegu Hanny University)
  • 장재식 (동국대학교 한의과대학 내과학교실) ;
  • 정지천 (동국대학교 한의과대학 내과학교실) ;
  • 신현철 (대구한의대학교 한의과대학 내과학교실)
  • Published : 2007.12.25

Abstract

Peroxynitrite ($ONOO^-$) is a reactive oxidant formed from superoxide anion radical (${\cdot}\;O_2-$) and nitric oxide (NO), which can oxidize cellular components such as essential protein, non-protein thiols, DNA, low-density lipoproteins and membrane phospholipids. ${\cdot}\;O_2-$ and $ONOO^-$ have contributed to the pathogenesis of diseases such as stroke, heart disease, Alzheimer's disease and atherosclerosis. Because of damaging effects of ${\cdot}\;O_2-$ and $ONOO^-$ oxidants, Vespae Nidus, which has been known to strengthen the kidneys to preserve the vital energy. was tested as a potential specific scavenger of those oxidants. In this study, the viability of Vespae Nidus (1, 10, 50 g/ml) to scavenge ${\cdot}\;O_2-$, NO, $ONOO^-$ and so to protect cells against tert-butylhydroxyperoxide (t-BHP) induced cell death was tested. The levels of ${\cdot}\;O_2-$ and $ONOO^-$ were detected by staining with DCFH-DA and DHR 123, respectively. Protein expression levels of COX-2, iNOS and $NF{-\kappa}B$ were assayed by western blot. Vespae Nidus blocked t-BHP-induced cell death in a dose-dependent fashion. Vespae Nidus inhibited t-BHP-induced production of ${\cdot}\;O_2-$, NO and $ONOO^-$ in YPEN cells. The lipid peroxide level was increased and glutathione level was decreased in lipopolysaccharide (LPS)-treated ICR mouse, whereas the ones in the Vespae Nidus-administered group were regulated beneficially. Vespae Nidus inhibited the expression of COX-2, iNOS and NF-κB (p65 and p50) genes in LPS-treated ICR mouse. The present study suggests that Vespae Nidus is a powerful antioxidant and promotes cellular defense activity by scavenging the toxic oxidants such as ${\cdot}\;O_2-$ and $ONOO^-$.

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. 平田結善緖. NO and NOS. 內分泌, 糖尿病科, 2: 207-214, 1996
  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. Chung, H.Y., Kim, H.J., Jung, K.J., Yoon, J.S., Yoo, M.A., Kim, K.W., Yu, B.P. The inflammatory process in aging. Reviews in Clinical Gerontology. 10: 207-222, 2000 https://doi.org/10.1017/S0959259800010327
  5. Chung, H.Y., Kim, H.J., Kim, K.W., Chio, J.S., Yu, B.P. Molecular inflammation hypothesis of aging based on the anti-aging mechanism of calorie restriction. Micro Res Techinq. 59: 264-272, 2002 https://doi.org/10.1002/jemt.10203
  6. Haenen, G.P.M.M., Paquay, J.B.G., Korthouwer, R.E.M., Bast, A. Peroxynitrite scavenging by flavonoids. Biochem Biophy Res Commu. 236: 591-596, 1997 https://doi.org/10.1006/bbrc.1997.7016
  7. Pannala, A.S., Rice-Evans, C.A., Halliwell, B., 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
  8. Kim, H.J., Kim, K.W., Yu, B.P., Chung, H.Y. The effect of age on cyclooxygenase-2 gene expression : NF-kB activation and IkB$\alpha$ degradation. Free Radical Biol Med. 28: 683-692, 2000 https://doi.org/10.1016/S0891-5849(99)00274-9
  9. Ghost, S., May, M.J., Kopp, E.B. NF-kB and Rel proteins : evolutionarity conserved mediators of immune responses. Annu Rev Immunol. 16: 225-260, 1998 https://doi.org/10.1146/annurev.immunol.16.1.225
  10. Korhonen, P., Helenius, M., Salminen, A. Age-related changes in the regulation 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. 전국 한의과대학 본초학교수. 本草學. 서울,영림사, pp 647-648, 1991
  12. 李尙仁. 本草學. 서울, 醫藥社, pp 534-535, 1983
  13. 江蘇新醫學院 編. 中藥大辭典. 上海, 上海科學技術出版社, pp 2736-2737, 1983
  14. 吳儀洛. 本草從新. 上海, 上海科學技術出版社, p 340, 1982
  15. 朱良春 編著. 蟲類藥的應用. 山西, 山西科學技術出版社, pp 58-70, 1994
  16. Scheller, S., Witczok, T., Imielski, S., Krol, W., Gabrys, J. and Shani, J. Free radical scavenging by ethanol extract of propolis. Int J Radiat Biol. 57(3):461-465, 1990 https://doi.org/10.1080/09553009014552601
  17. 양송원. 천연물질의 항균 활성에 관한 연구. 전남대학교 대학원, 2001
  18. 송윤선. Propolis의 약물학적 특성. 항염증 및 에스트로젠성 효과. 숙명여자대학교 대학원, 2001
  19. 허홍열. Propolis가 마우스에서 종양 발생 및 비장세포와 대식세포의 성장에 미치는 영향. 경북대학교 대학원, 1998
  20. 김영대, 이명종. 露蜂房의 항산화 작용에 관한 연구. 한방재활의학회지 13(1):11-22, 2003
  21. Tada, H., Shiho, O., Kuroshima, K., Koyama, M., Tsukamoto, K. An improved colorimetric assay for interleukin 2. J Immunol Methods. 93: 157-165, 1986 https://doi.org/10.1016/0022-1759(86)90183-3
  22. Cathcart, R., Schwiers, E., 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
  23. Kooy, N.W., Royall, J.A., Ischiropoulos, H., Beckman, J.S. Peroxynitrite-mediated oxidation of dihydrorhodamine 123. Free Radic Res Commun. 16: 149-156, 1994
  24. Nagata, N., Momose, K., 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
  25. Ohkawa, H., Ohishi, N., Yaki, K. Assay for lipid peroxide in animal tissues by thiobarbituric acid reaction. Anal Biochem. 95: 351-358, 1979 https://doi.org/10.1016/0003-2697(79)90738-3
  26. Ellman, G.L. Tissue sulfhydryl group. Arch Biochem Biophys. 82: 70-77, 1959 https://doi.org/10.1016/0003-9861(59)90090-6
  27. Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. Protein measurement with folin phenol reagent. J Biol Chem. 193: 265-275, 1951
  28. A Monti, M., Berti, E., Carminati, G. and Cusini, M. Occupational and cosmetic dermatitis from propolis. Contact Dermatitis. 9: 163-164, 1983
  29. Dobrowolski, J.W., Vohora, S.B., Sharma, K., Shah, S.A., Naqvi, S.A.H., Dandiya, P.C. Antibacterial, antifungal, antiamoebic, antiinflammatory and antipyretic studies on propolis bee products. J Ethnopharmacol. 35: 77-82, 1991 https://doi.org/10.1016/0378-8741(91)90135-Z
  30. Frenkel, K., Wei, H., Bhimani, R., Ye, J., Zaduanisky, J.A., Huamg, M., Ferraro, T., Conney, A.H., Grunberger, D. Inhibition of tumor promoter-mediated processes in mouse skin and bovine lens by caffeic acid phenethylester. Cancer Res. 53: 1255-12561, 1993
  31. Pascual, C., Gonzalez, R., Toricella, R.G. Scavenging action of propolis extract against oxygen radicals. J Ethnopharmacol. 41: 9-13, 1994 https://doi.org/10.1016/0378-8741(94)90052-3
  32. 정일윤. 프로폴리스에서 분리한 플라보노이드 화합물의 항산 화 활성 및 방사선 방어 효과. 한국식품영양학회지 34(2):162-166, 2005
  33. Choi, J.S., Chung, H.Y., Kang, S.S., Jung, M.J., Kim, J.W., No, J.K. and Jung, H.A. The Structure-Activity Relationship of Flavonoids as Scavengers of Peroxynitrite. Phytother. Res. 16: 232-235, 2002 https://doi.org/10.1002/ptr.828
  34. Kim, H.J., YU, B.P., Chung, H.Y. Moleculor inflammation hypothesis of aging based in the anti-aging mechanism of calorie restriction. Free Radical Biol Med. 10: 991-1005, 2002
  35. Hornsby, P.J., Crivello, J.F. The role of lipid peroxidation and biological antioxidants in the function of the adrenal cortex. Part 2. Mol Cell Endocrinol. 30: 123-147, 1983 https://doi.org/10.1016/0303-7207(83)90043-6
  36. Ross, D. Glutathione, free radicals and chemotherapeutic agents : Mechanism of free radical induced toxicity and glutathione dependent protection. Pharmacol Ther. 37(2):231-239, 1988 https://doi.org/10.1016/0163-7258(88)90027-7