Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of Resveratrol on the Induction of Cdk Inhibitor p21 and Pro-apoptotic Bax Expression by amyloid-β in Astroglioma C6 Cells

신경교 세포에서 resveratrol이 amyloid-β에 의해 유도되는 Cdk inhibitor p21 및 Bax 발현의 감소 효과

  • Kim Young Ae (Institute of Natural Medicine, Toyama Medical and Pharmaceutical University) ;
  • Lim Sun-Young (Division of Ocean Science, Korea Maritime University) ;
  • Ko Woo Shin (Department of Oriental Medicine, Dongeui University College of Oriental Medicine and Biomedical Research Center of Oriental Medicine) ;
  • Choi Byung Tae (Department of Oriental Medicine, Dongeui University College of Oriental Medicine and Biomedical Research Center of Oriental Medicine) ;
  • Lee Yong Tae (Department of Oriental Medicine, Dongeui University College of Oriental Medicine and Biomedical Research Center of Oriental Medicine) ;
  • Rhee Sook-Hee (Department of Food Science and Nutrition, Pusan National University) ;
  • Park Kun-Young (Department of Food Science and Nutrition, Pusan National University) ;
  • Lee Won-Ho (Department of Biology, Pusan National University) ;
  • Choi Yung Hyun (Department of Oriental Medicine, Dongeui University College of Oriental Medicine and Biomedical Research Center of Oriental Medicine)
  • 김영애 (일본 부산의과약과대학 화한약연구소) ;
  • 임선영 (한국해양대학교 해양과학기술대학 해양과학부) ;
  • 고우신 (동의대학교 한의과대학 한의학과 및 한방바이오연구센터) ;
  • 최병태 (동의대학교 한의과대학 한의학과 및 한방바이오연구센터) ;
  • 이용태 (동의대학교 한의과대학 한의학과 및 한방바이오연구센터) ;
  • 이숙희 (부산대학교 생활환경대학 식품영양학과) ;
  • 박건영 (부산대학교 생활환경대학 식품영양학과) ;
  • 이원호 (부산대학교 자연과학대학 생물학과) ;
  • 최영현 (동의대학교 한의과대학 한의학과 및 한방바이오연구센터)
  • Published : 2005.04.01
Download PDF
⟨ Previous Next ⟩

Abstract

Resveratrol (3,4',5-trihydroxy-trans-stilbene), a phytoalexin found in grape skins, peanuts, and red wine, has been reported to have a wide range of biological and pharmacological properties. $Amyloid-\beta$ deposition and senile plaque-associated astrocytes are common neuropathological features of Alzheimer's disease. In this study, we have explored the effects of resveratrol on $amyloid-\beta-peptide-mediated$ cytotoxicity in vitro and modulation of cell growth-regulatory gene products in astroglioma C6 cells to elucidate its possible mechanism for anti-cytotoxicity. Exposure of C6 cells to $Amyloid-\beta$ resulted in dose-dependent growth inhibition and morphological changes of C6 cells, which were recovered by pre-treatment with resveratrol. The anti-proliferative effect of $amyloid-\beta$ was associated with the induction of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21 (WAF1/CIP1) expression assessed by RT-PCR and Western blot analysis in time-dependent manner in C6 cells. In addition, the pro-apoptotic Bax expression was also up-regulated in $amyloid-\beta-treated$ C6 cells without alteration of anti-apoptotic Bcl-2 and $Bcl-X_L$ expression. However, pre-treatment of resveratrol significantly inhibited $amyloid-\beta-induced$ p53, p21 and Bax levels, suggesting that the modulation of p53, p21 and Bax levels could be one of the possible pathways by which resveratrol functions as anti-cytotoxic agent. Our results demonstrate that resveratrol may enhance the protection against $amyloid-\beta-induced$ cytotoxicity by promoting the survival of glial cells.

Resveratrol (3,4',5-trihydroxy-trans-stilbene)은 포도와 같은 식물에서 각종 감염균으로부터 자신의 몸을 보호하기 위하여 생성되는 물질인 phytoalexin의 일종으로 강력한 항산화작용, 암예방 효과 및 항암 작용을 포함한 각종 약리작용을 가진 것으로 보고 되어져 오고 있다. Alzheimer 환자의 뇌에 축적되어 뇌 신경세포를 죽이는 amyloid plaque의 주 성분은 $amyloid-\beta$의 축적에 의한 것인데, $amyloid-\beta$는 정상적인 단백질 신진대사 과정의 결과로 체내 모든 세포들로부터 생성되는 물질이다. 본 연구에서는 resveratrol의 세포독성 보호효과에 관한 효능을 검증하기 위하여 C6 신경교세포에서 $amyloid-\beta-peptide$ (fragment 31-35)에 의한 세포독성 및 세포성장 조절관련 주요 유전자들의 발현에 미치는 resveratrol의 영향을 조사하였다. $Amyloid-\beta$가 처리된 C6세포는 처리 농도의존적으로 증식이 억제되었으며, 형태적 변형도 유발 되었으나 resveratrol의 전처리에 의하여 효과적으로 차단되었다. RT-PCR 및 Western blot analysis에 의한 결과에서 $amyloid-\beta$ 처리에 의한 세포증식 억제는 종양억제유전자 p53 및 Cdk 억제제인 p21 (WAF1/CIP1) 발현이 증가되었다. 또한 apoptosis 유발에 매우 중요한 역할을 수행하는 Bax의 발현도 $amyloid-\beta$가 처리된 C6 세포에서 발현이 증가되었으나 apoptosis 유발억제에 관여하는 Bcl-2및 $Bcl-X_{L}$ 발현에는 큰 영향을 미치지 못하였다. 그러나 resveratrol이 전처리된 세포에서는 처리 농도 의존적으로 $amyloid-\beta$에 의해 유도되는 p53, p21 및 Bax의 발현이 정상수준으로 회복되었다.

Keywords

References

  1. Barger, S.W. and A.D. Harmon. 1997. Microglial activation by Alzheimer amyloid precursor protein and modulation by apolipoprotein E. Nature 388, 878-881 https://doi.org/10.1038/42257
  2. Bhat, K.P.L., J.W. Kosmeder 2nd and J.M. Pezzuto. 2001. Biological effects of resveratrol. Antioxid. Redox Signal 3, 1041-1064 https://doi.org/10.1089/152308601317203567
  3. Bhavnani, B.R., A. Cecutti, A. Cerulath, A.C. Woolever and M. Berco. 2001. Comparison of the antioxidant effects of equine estrogens, red wine components, vitamin E, and probucol on low-density lipoprotein oxidation in postmenopausal women. Menopause 8, 408-419 https://doi.org/10.1097/00042192-200111000-00005
  4. Chan, M.M. 2002. Antimicrobial effect of resveratrol on dermatophytes and bacterial pathogens of the skin. Biochem. Pharmacol. 63, 99-104 https://doi.org/10.1016/S0006-2952(01)00886-3
  5. Chiarugi, V., L. Magnelli and G. Basi. 1994. Apoptosis and the cell cycle. Cell. Mol. Biol. Res. 40, 603-612
  6. Choi, Y.H., KR. Kong, Y.A. Kim, K.O. Jung, J.H. J.H. Kil, S.H. Rhee and K.Y. Park. 2003. Induction of Bax and activation of caspases during $\beta$-sitosterol-mediated apoptosis in human colon cancer cells. Int. J. Oncol. 23, 1657-1661
  7. Choi, Y.H., S.J. Lee, P. Nguyen, J.S.. Jang, J. Lee, M.L. Wu, E. Takano, M. Maki, PA Henkart and J.B. Trepel. 1997. Regulation of eyclin D1 by cal pain protease. J. Biol. Chem. 272, 28479-28484 https://doi.org/10.1074/jbc.272.45.28479
  8. Draczynska-Lusiak, B., Y.M. Chen and A.Y. Sun. 1998. Oxidized lipoproteins activate NF-$\kappa$B binding activity and apoptosis in PC12 cells. Neuroreport 9, 527-532 https://doi.org/10.1097/00001756-199802160-00028
  9. El Khoury, J., S.E. Hickman, C.A. Thomas, L. Cao, S.C Silverstein and J.D. Loike. 1996. Scavenger receptor-mediated adhesion of microglia to beta-amyloid fibrils. Nature 382, 716-719 https://doi.org/10.1038/382716a0
  10. El-Deiry, W.S., T. Tokino, V.E. Velculesco, D.B. Levy, R. Parsons, J.M. Trent, D. Lin, E.W. Mercer, KW. Kinzler and B. Vogelstain. 1993. WAF1, a potential mediator of p53 tumor suppression. Cell 75, 817-825 https://doi.org/10.1016/0092-8674(93)90500-P
  11. Ciulian, D., L.J. Haverkamp, J.H. Yu, W. Karshin, D. Tom, J. Li, J. Kirkpatrick, L.M. Kuo and A.E. Roher. 1996. Specific domains of $\beta$-amyloid from Alzheimer plaque elicit neuron killing in human microglia. J. Neurosci. 16, 6021-6037
  12. Goldberg, D.M., S.E. Hahn and J.G. Parkes. 1995. Beyond alcohol: beverage consumption and cardiovascular mortality. Clin. Chim. Acta. 237, 155-187 https://doi.org/10.1016/0009-8981(95)06069-P
  13. Hadi, S.M., S.F. Asad, S. Singh and A. Ahmad. 2000. Putative mechanism for anticancer and apoptosis-inducing properties of plant-derived polyphenolic compounds. IUBMB Life 50, 167-171 https://doi.org/10.1080/152165400300001471
  14. Hansson, E. 1998. Astroglia from defined brain regions as studied with primary cultures. Prog. Neurobiol. 30, 369-397
  15. Harder, D.R., C. Zhang and D. Gebremedhin. 2002. Astrocytes function in matching blood flow to metabolic activity. News Physiol. Sci. 17, 27-31
  16. Hockenbery, D.M., Z.M. Oltvai, X.M. Yin, CL. Milliman and S.J. Korsmeyer. 1993. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75, 241-251 https://doi.org/10.1016/0092-8674(93)80066-N
  17. Igura, K, T. Ohta, Y. Kuroda and K Kaji. 2001. Resveratrol and quercetin inhibit angiogenesis in vitro. Cancer Lett. 171, 11-16 https://doi.org/10.1016/S0304-3835(01)00443-8
  18. Jang, D.S., B.S. Kang, S.Y. Ryu, J.M. Chang, KR. Min and y. Kim. 1999. Inhibitory effects of resveratrol analogs on unopsonized zymosan-induced oxygen radical production. Biochem. Pharmacol. 57, 705-712 https://doi.org/10.1016/S0006-2952(98)00350-5
  19. Jang, J.H. and Y.J. Surh. 2001. Protective effects of resveratrol on hydrogen peroxide-induced apoptosis in rat pheochromocytoma (PC12) cells. Mutat. Res. 496, 181-190 https://doi.org/10.1016/S1383-5718(01)00233-9
  20. Johnson, S.A., G.M. Pasinetti, P.C. May, P.A. Ponte, B. Cordell and C.E. Finch. 1998. Selective reduction of mRNA for the $\beta$-amyloid precursor protein that lacks a Kunitztype protease inhibitor motif in cortex from Alzheimer brains. Exp. Neurol. 102, 264-826 https://doi.org/10.1016/0014-4886(88)90104-5
  21. Kopan, R. and A. Goate. 2002. Aph-2/Nicastrin: an essential component of gamma-secretase and regulator of Notch signaling and Presenilin localization. Neuron 33, 321-324 https://doi.org/10.1016/S0896-6273(02)00585-8
  22. Kosik, K.S. 1992. Alzheimer's disease: a cell biological perspective. Science 256, 780-783 https://doi.org/10.1126/science.1589757
  23. Kvamme, E., LA. Torgner and B. Roberg. 2001. Kinetics and localization of brain phosphate activated glutaminase. J. Neurosci. Res. 66, 951-958 https://doi.org/10.1002/jnr.10041
  24. Lowe, S.W., H.E. Ruley, T. Jacks and D.E. Housman. 1993. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74, 957-967 https://doi.org/10.1016/0092-8674(93)90719-7
  25. Lue, L.F., D.G. Walker and J. Rogers. 2001. Modeling microglial activation in Alzheimer's disease with human poshnortem microglial cultures. Neurobiol. Aging 22, 945-956 https://doi.org/10.1016/S0197-4580(01)00311-6
  26. Mahley, R.W. and S.C. Rall Jr. 2000. Apolipoprotein E: far more than a lipid transport protein. Annu. Rev. Genomics Hum. Genet. 1, 507-537 https://doi.org/10.1146/annurev.genom.1.1.507
  27. Marin, D.B., M.C. Sewell and A. Schlechter. 2002. Alzheimer's disease. Accurate and early diagnosis in the primary care setting. Geriatrics 57, 36-40
  28. Miyashita, T. and J.C. Reed. 1995. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80, 293-299 https://doi.org/10.1016/0092-8674(95)90412-3
  29. Morgan, D.O. 1995. Principles of CDK regulation. Nature 374, 131-134 https://doi.org/10.1038/374131a0
  30. Potter, H., L.M. Wefes and L.N. Nilsson. 2001. The inflammation-induced pathological chaperones ACT and apo-E are necessary catalysts of Alzheimer amyloid formation. Neurobiol. Aging 22, 923-930 https://doi.org/10.1016/S0197-4580(01)00308-6
  31. Reddy, B.S., C.X. Wang, H. Samaha, R. Lubet, V.E. Steele, G,J. Kelloff and C.V. Rao. 1997. Chemoprevention of colon carcinogenesis by dietary perillyl alcohol. Cancer Res. 57, 420-425
  32. Sherr, C.J., 2000. The Pezcoller lecture: cancer cell cycles revisited. Cancer Res. 60, 3689-3695
  33. Soleas, G.J., E.P. Diamandis and D.M. Goldberg. 1997. Wine as a biological fluid: history, production, and role in disease prevention. J. Clin. Lab. Anal. 11, 287-313 https://doi.org/10.1002/(SICI)1098-2825(1997)11:5<287::AID-JCLA6>3.0.CO;2-4
  34. Sun, N.J., S.H. Woo, J.M. Cassady and R.M. Snapka. 1998. DNA polymerase and topoisomerase II inhibitors from Psoralea corylifolia. J. Nat. Prod. 61, 362-366 https://doi.org/10.1021/np970488q
  35. Surh, Y.J., KS. Chun, H.H. Cha, S.S. Han, Y.S. Keum, KK Park and SS. Lee. 2001. Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-$\kappa$B activation. Mutat. Res. 480-481, 243-268
  36. Wang, Z., Y. Huang, J. Zou, K Cao, Y. Xu and J.M. WU. 2002. Effects of red wine and wine polyphenol resveratrol on platelet aggregation in vivo and in vitro. Int. J. Mol. Med. 9, 77-79
  37. Wu, J.M., Z.R. Wang, T.C. Hsieh, J.L. Bruder, J.G. Zou and Y.Z. Huang. 2001. Mechanism of cardioprotection by resveratrol, a phenolic antioxidant present in red wine. Int. J. Mol. Med. 8, 3-17
  38. Yan, S.D., X. Chen, J. Fu, M. Chen, H. Zhu, A. Roher, T. Slattery, L. Zhao, M. Nagashima, J. Morser, A. Migheli, P. Nawroth, D. Stern and A.M. Schmidt. 1996. RAGE and arnyloid--$\beta$ peptide neurotoxicity in Alzheimer's disease. Nature 382, 685-691 https://doi.org/10.1038/382685a0
  39. Zou, J., Y. Huang, Q. Chen, E. Wei, K Cao and J.M. Wu. 2000. Effects of resveratrol on oxidative modification of human low density lipoprotein. Chin. Med. J. (Engl) 113, 99-102