한방재활의학과학회지 (Journal of Korean Medicine Rehabilitation)

  • 제24권2호
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  • Pages.41-50
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  • 2014
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  • 1229-1854(pISSN)
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  • 2288-4114(eISSN)
한방재활의학과학회 (The society of Korean Medicine Rehabilitation)
권호 표지

위령선 추출물이 Human Neuroblastoma 세포주에서 $CoCl_2$에 의해 유도된 세포사멸에 미치는 보호효과

Protective Effect of Clematidis Radix Extract on $CoCl_2$-induced Apoptosis in Human Neuroblastoma Cells

  • 박정우 (가천대학교 한의과대학 한방재활의학과교실) ;
  • 임형호 (가천대학교 한의과대학 한방재활의학과교실)
  • Park, Jung-Woo (Department of Korean Medicine Rehabilitation, College of Korean Medicine, Gachon University) ;
  • Lim, Hyung-Ho (Department of Korean Medicine Rehabilitation, College of Korean Medicine, Gachon University)
  • 투고 : 2014.03.18
  • 심사 : 2014.03.28
  • 발행 : 2014.04.30
⟨ 이전 논문 다음 논문 ⟩

초록

Objectives The purpose of this study was to evaluate the effects of Clematidis radix extract on $CoCl_2$-induced apoptosis in SH-SY5Y human neuroblastoma cells. Methods In order to investigate the protective effect of Clematidis radix on $CoCl_2$-induced cytotoxicity in neuronal cells, MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, DAPI(4,6-diamidino-2-phenylindoleI) staining, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) assay, DNA fragmentation assay and western blotting were performed on SH-SY5Y human neuroblastoma cells. Results Cells treated with $CoCl_2$ exhibited several apoptotic features, while cells pre-treated with Clematidis radix prior to $CoCl_2$ exposure showed a decrease in the occurrence of apoptotic features. $CoCl_2$ increased HIF-$1{\alpha}$ expression, in contrast, Clematidis radix treatment decreased $CoCl_2$-induced HIF-$1{\alpha}$ expression. Pre-treatment with the extract of Clematidis radix suppressed Bax, cytochrome c, and caspase-3 expressions, and also increased Bcl-2 expression in SH-SY5Y human neuroblastoma cells. Conclusions These results suggest that Clematidis radix may exert a protective effect on $CoCl_2$-induced apoptosis in SH-SY5Y human neuroblastoma cells.

키워드

참고문헌

  1. An SM, Lee SI, Choi SW, Moon SW, Boo YC. p- Coumaric acid, a constituent of Sasa quelpaertensis Nakai, inhibits cellular melanogenesis stimulated by $\alpha$- melanocyte stimulating hormone. Br J Dermatol. 2008; 159(2):292-9. https://doi.org/10.1111/j.1365-2133.2008.08653.x
  2. Yu s, Keum DH, Lee MJ. Effect of Clematidis Radix on Anti-Inflammatory, Analgesic and Antipyretic in Animal Model. J Oriental Rehab Med. 2004;14(2):75-84.
  3. Song SW, Song TW. Immunity Responses of the Clematidis Radix to the Synovial Cells Isolated from Patients with Rheumatoid Arthritis. Graduate School, Daejeon University. 2003:1-50.
  4. Lee JS, Kim KH, Lee SD, Kim KS. The Effect of Clematidis Radix Herbal-acupuncture Solution, on Collagen, Adjuvant, Lipopoly-saccharide and Phospholipase A2 Induced Rheumatoid Arthritis in Mice. JKAMS. 2012;29(1):127-37.
  5. Choi WJ, Kim KB, Kim KH, Kim JH. Analgesic Effect of Clematidis Radix Herbal acupuncture on the Test Rats with Induced Acute Pain. JKAMS. 2009;26(6):91-100.
  6. Song YH, Yoo JE, Lim HJ, Yoo DY. Effects of Clematidis Radix Extract on Osteoclastogenesis and Gene Expression in RANKL -induced RAW 264.7 Cell. J Oriental Obstetrics & Gynecology. 2010;23(3):78-90.
  7. Chen JX, Zhao T, Huang DX. Protective effects of edaravone against cobalt chloride-induced apoptosis in PC12 cells. Neurosci Bull. 2009;25(2):67-74. https://doi.org/10.1007/s12264-009-1215-6
  8. Guglielmotto M, Tamagno E, Danni O. Oxidative stress and hypoxia contribute to Alzheimer's disease pathogenesis: two sides of the same coin. Scientific World Journal. 2009;9:781-91. https://doi.org/10.1100/tsw.2009.93
  9. Wang G, Hazra TK, Mitra S, Lee HM, Englander E. Mitochondrial DNA damage and a hypoxic response are induced by CoCl2 in rat neuronal PC12 cells. Nucleic Acids Res. 2000;28(10):2135-40. https://doi.org/10.1093/nar/28.10.2135
  10. Minet E, Arnould T, Michel G, Roland I, Mottet D, Raes M, Remacle J, Michiels C. ERK activation upon hypoxia: involvement in HIF-1 activation. FEBS Lett. 2000;468(1):53-8. https://doi.org/10.1016/S0014-5793(00)01181-9
  11. Shin KS, Kwon GR, Kim JW, Son NW, Jeong SH, Shin GC, Lee WC. A Study for Neuroproctective Effect of Bupleuri Radix on BV-2 Microglia Cells. J The Korea Institute Of Oriental Medical Informatics. 2004;10(2):61-69.
  12. Park BY, Song YK, Lim HH. Protective Effect of Acanthopanax Cortex Extract on CoCl2-induced Apoptosis in Human SK-N-MC Neuroblastoma Cells. J Oriental Rehab Med. 2012;22(2):1-16.
  13. Sung MS, Song YK, Lim HH. Protective Effect of Sorbus Commixta Cortex Extract on CoCl2-induced Apoptosis in Neuroblastoma Cell Line SK-N-MC. J Oriental Rehab Med. 2012;22(2):17-29.
  14. Hu YS, Lee SG, Chai YG, Jung KH, Kim JH. Gene expression profiling of SH-SY5Y cells in neuroprotective effect of total ginsenosides on H202 induced neurotoxicity. J Oriental Neuro-psychiatry. 2007;18(1):95-110.
  15. Bae KR, Doo AR, Kim SN, Park JY, Park HJ, Lee HJ, Kwon KR. Neuroprotective Effects of Bee Venom, which Removes High Molecular Elements against MPP+-induced Human Neuroblastoma SH-SY5Y Cell Death. J Korean Oriental Internal Med. 2010;31(2):254-63.
  16. 전국한의과대학 공동교재편찬위원회. 본초학. 서울:영림사. 2011:305-6.
  17. An SM, Lee SI, Choi SW, Moon SW, Boo YC. p-Coumaric acid, a constituent of Sasa quelpaertensis Nakai, inhibits cellular melanogenesis stimulated by $\alpha$- melanocyte stimulating hormone. Br J Dermatol. 2008; 159(2):292-9. https://doi.org/10.1111/j.1365-2133.2008.08653.x
  18. Zeno S, Zaaroor M, Leschiner S, Veenman L, Gavish M. CoCl2 induces apoptosis via the 18 kDa translocator protein in U118MG human glioblastoma cells. Biochemistry. 2009;48(21):4652-61. https://doi.org/10.1021/bi900064t
  19. Zou H, Henzel WJ, Liu X, Lutschg A, Wang X. Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell. 1997;90(3):405-13. https://doi.org/10.1016/S0092-8674(00)80501-2
  20. Spector MS, Desnoyers S, Hoeppner DJ, Hengartner MO. Interaction between the C. elegans cell-death regulators CED-9 and CED-4. Nature. 1997;385(6617):653-6. https://doi.org/10.1038/385653a0
  21. Wolter KG, Hsu YT, Smith CL, Nechushtan A, Xi XG & Youle RJ. Movement of Bax from the cytosol to mitochondria during apoptosis. J Cell Biol. 1997;139(5):1281-92. https://doi.org/10.1083/jcb.139.5.1281
  22. Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature. 1998;391(6662):43-50. https://doi.org/10.1038/34112
  23. Wyllie AH. Glucocorticoid induced thymocytes apoptosis is associated with endogenous endonuclease activation. Nature. 1980;284(5756):555-6. https://doi.org/10.1038/284555a0
  24. Gavrieli Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992;119(3):493-501. https://doi.org/10.1083/jcb.119.3.493
  25. Sleiman RJ, Catchpoole DR, Stewart BW. Drug-induced death of leukaemic cells after G2/M arrest: higher order DNA fragmentation as an indicator of mechanism. Br J Cancer. 1998;77(1):40-50.
  26. Ema M, Hirota K, Mimura J, Abe H, Yodoi J, Sogawa K, Poellinger L, Fujii-Kuriyama Y. Molecular mechanisms of transcription activation by HLF and HIF1$\alpha$ in response to hypoxia: their stabilization and redox signal-induced interaction with CBP/p300. EMBO J. 1999;18(7):1905-14. https://doi.org/10.1093/emboj/18.7.1905
  27. Greijer AE, van der Wall E. The role of hypoxia inducible factor 1(HIF-1) in hypoxia induced apoptosis. J Clin Pathol. 2004;57(10):1009-14. https://doi.org/10.1136/jcp.2003.015032
  28. Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 1993;74(4):609-19. https://doi.org/10.1016/0092-8674(93)90509-O
  29. Harris MH, Thompson CB. The role of Bcl-2 family in the regulation of outer mitochondrial membrane permeability. Cell Death Differ. 2000;7(12):1182-91. https://doi.org/10.1038/sj.cdd.4400781
  30. Lim ML, Lum MG, Hansen TM, Roucou X, Nagley P. On the release of cytochrome c from mitochondria during cell death signaling. J Biomed Sci. 2002;9(6 Pt 1):288-506. https://doi.org/10.1007/BF02256075
  31. Shimizu S, Tsujimoto Y. Proapoptotic BH3-only Bcl-2 family members induce cytochrome c release, but not mitochondrial membrane potential loss, and do not directly modulate voltage-dependent anion channel activity. Proc Natl Acad Sci USA. 2000;97(2):577-82. https://doi.org/10.1073/pnas.97.2.577
  32. Jü.rgensmeier JM, Xie Z, Deveraux Q, Ellerby L, Bredesen D, Reed JC. Bax directly induces release of cytochrome c from isolated mitochondria. Proc Natl Acad Sci USA. 1998;95(9):4997-5002. https://doi.org/10.1073/pnas.95.9.4997
  33. Boatright KM, Salvesen GS. Mechanisms of caspase activation. Curr Opin Cell Biol. 2003;15(6):725-31. https://doi.org/10.1016/j.ceb.2003.10.009
  34. Araya R, Uehara T & Nomura Y. Hypoxia induces apoptosis in human neuroblastoma SK-N-MC cells by caspase activation accompanying cytochrome c release from mitochondria. FEBS Lett. 1998;439(1-2):168-72. https://doi.org/10.1016/S0014-5793(98)01363-5