Effects of Daechilgi-tang on Glutamate-induced Apoptosis in C6 Glial Cells

대칠기탕(大七氣湯)이 Glutamate에 의한 C6 Glial 세포의 Apoptosis에 미치는 영향

  • Kim, Hye-Yoon (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Ko, Seok-Jae (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Bang, Chang-Ho (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Shin, Sun-Ho (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Lee, John Dong-Yeop (Dept. of Third Medicine, Professional Graduate School of Oriental Medicine, Won-Kwang University) ;
  • Lee, In (Dept. of Internal Medicine, College of Oriental Medicine, Pu-San National University)
  • 김혜윤 (원광대학교 심계내과학교실) ;
  • 고석재 (원광대학교 심계내과학교실) ;
  • 방창호 (원광대학교 심계내과학교실) ;
  • 신선호 (원광대학교 심계내과학교실) ;
  • 이동엽 (원광대학교 한의학전문대학원 제3의학과) ;
  • 이인 (부산대학교한의학전문대학원)
  • Published : 2010.12.30

Abstract

Objectives : The water extract of Daechilgi-tang(DCGT) has traditionally been used for treatment of qi stagnation(氣滯), which is considered to be one of the important causes of neuronal disease in oriental medicine. However, little is known about the mechanism by which DCGT protects neuronal cells from brain cell damages. Methods and Results : The author tested the mechanism of the cytoprotective effect of DCGT on glutamate -stimulated rat C6 glial cells. DCGT significantly protected C6 glial cells from glutamate in MTT assay. Pre-treatment of C6 glial cells with DCGT markedly inhibited the DNA fragmentation of C6 cells induced by glutamate. Glutamate increased the generation of reactive oxygen species(ROS) and intracellular calcium level in C6 glial cells. However, pre-treatment with DCGT markedly suppressed the increase of ROS generation and intracellular calcium accumulation induced by glutamate. Among apoptosis signaling mediators, DCGT markedly increased the expression level of Bcl2 in glutamate-treated cells. It also inhibited the cleavage of caspase-3 and PARP proteins by glutamate in C6 glial cells. Conclusions : These results suggest that DCGT protects brain cells from glutamate cytotoxicity through inhibition of ROS generation and activation of apoptosis signaling pathway as well as induction of the anti-oxidant system.

References

  1. 李梴. 國譯編註醫學入門. 서울: 南山堂; 2001, p. 403.
  2. 黃道淵. 方藥合編. 서울: 永林社; 2002, p. 278.
  3. 전국한의과대학심계내과학교실. 심계내과학. 서울: 군자출판사; 2006, p. 331-6.
  4. Peter L. Ischemic Cell Death in Brain Neurons. Physiol. Review. 1999;70:1499.
  5. 하지용, 정병억, 이선구, 윤병길. 大七氣湯이 抗癌, 免疫調節作用 및 apoptosis에 미치는 影響. 동의생리병리학회지. 2001;15(1):111-24.
  6. 이승재. 大七氣湯이 자궁근종 세포의 증식억제, MAP kinase 활성 및 cell apoptosis에 미치는 影響. 경희대학교 박사학위논문. 2002.
  7. Arce MP, Rodriguez-Franco MI, Gonzalez-Munoz GC, Perez C, Lopez B, Villarroya M, Lopez MG et al. Neuroprotective and Cholinergic Properties of Multifunctional Glutamic Acid Derivatives for the Treatment of Alzheimer's Disease. J. Med Chem. 2009;52(22):7249-57. https://doi.org/10.1021/jm900628z
  8. Straus SE, Majumdar SR, McAlister FA. New evidence for stroke prevention: scientific review. JAMA. 2002;288(11):1388-95. https://doi.org/10.1001/jama.288.11.1388
  9. 김세길. 風의 病理的 意味糾明과 中風의 原因 및 治療에 대한 東西醫學的 比較. 대한한의학회지. 1995;16(1):96-117.
  10. 尹用甲. 東醫方劑와 處方解說. 서울: 醫聖堂; 1998, p. 118.
  11. 윤상학, 정인철, 이상룡. 香附子가 치매병태 모델에 미치는 影響. 동의신경정신과학회지. 2004;14(1):59-74.
  12. 이금수, 정현우, 이원석. 靑皮가 腦血流力學에 미치는 실험적 연구. 동의생리병리학회지. 2002;16(1):104-10.
  13. 서운교, 정효원, 박용기. 생강 클로로포름 분획의 활성화된 腦神經膠細胞에서 염증반응 억제효과. 대한본초학회지. 2008;23(3):73-83.
  14. 정환용, 주혜진, 정혜미, 신우진, 서운교. 腦神經小膠細胞에서 생강 헥산 분획물의 염증매개물질 生成 抑制效果. 대한한의학회지. 2009;30(2):17-29.
  15. 오용열. 배양섬유모세포에서 산소유리기의 세포 독성에 대한 진피의 항산화효과에 관한 연구. 대한본초학회지. 2006;21(4):143-8.
  16. Landucci E, Boscia F, Gerace E, Scartabelli T, Cozzi A, Moroni F, Mannaioni G, Pellegrini-Giampietro DE. Involvement of endocannabinoid signaling in the neuroprotective effects of subtype 1 metabotropic glutamate receptor antagonists in models of cerebral ischemia. Int Rev Neurobiol. 2009;85:337-50. https://doi.org/10.1016/S0074-7742(09)85023-X
  17. 황인영, 신임철, 송연숙, 성민제, 박혜지, 박철범 등. Glutamate에 의한 세포 내 칼슘농도 변화와 세포독성과의 관계. Toxicological Research. 2002;18(4):355-62.
  18. 안종석, 김동희, 김윤식, 이용구, 남궁욱, 설인찬. 加味鷄血藤湯이 Glutamate receptor와 Free radical 및 뇌손상 보호에 미치는 영향. 동의생리병리학회지. 2003;17(3):728-37.
  19. 오병열, 김민창, 유병열, 최영, 설인찬 加味柴胡湯이 뇌허혈 시 Glutamate receptor, free radical 및 뇌손상 보호에 미치는 영향. 대한한의학회지. 2004;25(3):32-44.
  20. 강익현, 한상혁, 이인, 문병순. 歸脾湯이 Glutamate에 의한 C6 Glial Cell의 Apoptosis에 미치는 영향. 대한한의학회지. 2001;22(4):45-57.
  21. 정대영, 최철원, 문병순. 四物湯이 Glutamate에 의한 해마세포의 손상에 미치는 영향. 대한한의학회지. 2009;30(1):64-75.
  22. 정승원, 최철원, 김봉상, 문병순. 疎風湯이 Glutamate에 의한 C6 Glial Cell의 Apoptosis에 미치는 영향. 동의생리병리학회지. 2008;22(6):1423-30.
  23. Solary E, Droin N, Bettaieb A, Corcos L, Dimanche-Boitrel MT, Garrido C. Positive and negative regulation of apoptotic pathways by cytotoxic agents in hematological malignancies. Leukemia. 2000;14:1833-49. https://doi.org/10.1038/sj.leu.2401902
  24. Wilson JX, Gelb AW. Free radicals, antioxidants, and neurologic injury: possible relationship to cerebral protection by anesthetics. J. Neurosurg Anesthesiol. 2002;14:66-79. https://doi.org/10.1097/00008506-200201000-00014
  25. Hengartner MO. The biochemistry of apoptosis. Nature. 2000;407:770-6. https://doi.org/10.1038/35037710
  26. Deveraux QL, Schendel SL, Reed JC. Antiapoptotic proteins: the Bcl-2 and inhibitor of apoptosis protein families. Cardiol Clin. 2001;19:57-74. https://doi.org/10.1016/S0733-8651(05)70195-8