Effects of Samultang on Glutamate-Induced Apoptosis of Hippocampus Cells

사물탕(四物湯)이 Glutamate에 의한 해마세포의 손상에 미치는 영향

  • Jeong, Dae-Young (Department of Internal Medicine, School of Oriental Medicine, Wonkwang University) ;
  • Choi, Chul-Won (Department of Internal Medicine, School of Oriental Medicine, Wonkwang University) ;
  • Moon, Byung-Soon (Department of Internal Medicine, School of Oriental Medicine, Wonkwang University)
  • 정대영 (원광대학교 한의과대학 내과학교실) ;
  • 최철원 (원광대학교 한의과대학 내과학교실) ;
  • 문병순 (원광대학교 한의과대학 내과학교실)
  • Published : 2009.01.30

Abstract

Objective: This study was designed to investigate the effect of Samultang (SMT) under hippocampus cells ischemia both in vitro and in vivo. Methods: In the in vitro study, HT22 cells, predominantly detected in the cytoplasm, which coincides with the location of the mitochondria, were used as indicators. In the in vivo study, permanent middle cerebral artery occlusion (MCAO) was induced on rats. SMT was given orally 2 h before induction of permanent focal brain ischemic injury. Result: In the in vitro study, SMT had protective effects in glutamate-induced cytotoxicity, which was revealed as apoptosis characterized by chromatic condensation and the loss of mitochondrial membrane potential in HT22 cells. In the in vivo study, TTC (2,3,5-triphenyltetrazolium chloride) staining showed a marked ischemic injury in blood supply territory of the middle cerebral artery (MCA) such as the cerebral cortex and striatum. However, treatment with SMT significantly reduced infarcted volume. SMT increased marked survival of HT22 cells against glutamate-induced cytotoxicity in MTT assay. Conclusion: These results suggest that water extract of SMT provides neuroprotection against ischemic or oxidative injury by inhibition of apoptotic cell death.

Keywords

References

  1. 강성돈, 정진원,문병순,김종문. 뇌졸중의 역학적 동향에 대한 연구. 대한신경외과학학회지. 1999;28:509-13.
  2. American Heart Association. Heart and Stroke Facts Statistical Supplement. New York. American Heart Association. 1994:12.
  3. Kirino T, Tamura A, Sano K. Selective vulnerability of the hippocampus to ischemia-reversible and irreversible types of ischemic cell damage. Progress in Brain Res. 1985;63:39-58. https://doi.org/10.1016/S0079-6123(08)61974-3
  4. Kogure T, Koure K. Molecular and biochemical events within the brain subjected to cerebral ischemia. Clin Neurosci. 1997;4:179-83.
  5. Gray JA, McNaughton N. Comprasion between the behavioral effects of septal and hippocampal lesions. A review. Neuroscience and biobehavioral Reviews. 1982;7:119-88. https://doi.org/10.1016/0149-7634(83)90014-3
  6. Blozovski D. PA-Learning in young rats with dorsal hippocampus and hippocampoentorhinal atropine. Pharmacology. Biochemistry and Behavior. 1979;10:369-72. https://doi.org/10.1016/0091-3057(79)90199-0
  7. 陳師文. 太平惠民和劑局方. 臺北: 族風出版社. 1975:242.
  8. 朱震亨. 新編丹溪心法附餘(下卷). 서울:大成文化社. 1993:702-3.
  9. 許浚. 東醫寶鑑. 서울: 大成文化社. 1981:235.
  10. 김완희. 四物揚에 대한 고찰. 大韓韓醫學會誌. 1996:2:35-41.
  11. 陳偉. 方劑學. 서울: 醫聖堂. 1993:205-8.
  12. 尹用甲. 東醫方劑와 處方解說. 서울: 醫聖堂. 1998:163-70.
  13. 남창규. 四物揚이 血管內皮細胞에 미치는 影響. 圓光大學校 韓醫學專門大學院. 1998:24.
  14. 김도환. Nitric Oxide에 의해 유발된 C6 glial 세포독성에 대한 사물탕의 방어효과. 圓光大學校 韓醫學專門大學院. 2000:30.
  15. 조권일, 정승원, 장재호, 이대용, 박세욱, 이 인, 신선호, 문병순. 培養心筋細胞의 酸化的 慎傷에 대한 四物湯의 防禦效果. 大韓韓醫學會誌. 2005;26:174-86.
  16. 李哲浣 : 四君子湯, 四物湯 및 八物湯이 筋肉疲勞恢復에 미치는 實驗的 硏究. 慶喜大學校 大學院. 1988.
  17. 은재순. 사물탕이 L1210 세포 이식 및 항암제를 투여한 마우스의 면역세포에 미치는 영향. 생약학회지. 1998:113:110-9.
  18. 이성환, 임성우. 알코올로 유발된 생쥐의 위점막 손상에 대한 사물탕의 보호효과. 大韓韓醫學會誌. 2004;25:1-11.
  19. 金完熙, 崔達永. 臟腑辨蹬論治. 서울: 成輔社. 1988:58-9,160-5.
  20. Gerriets T, Li F, Silva MD, Meng X, Brevard M, Sotak CH, Fisher M. The microsphere model: evaluation of a new stroke model for permanent middle cerebral artery occlusion in rats. J Neurosci Methods. 2003;122:201-11. https://doi.org/10.1016/S0165-0270(02)00322-9
  21. Cole SP. Rapid chemosensitivity testing of human lung tumor cells using the MTT assay. Cancer Chemother Pharmacol. 1986;17:159-63.
  22. Herrmann M, Lorenz HM, Voll R, Grunke M, Woith W, Kalden JR. A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Research. 1994;22:5506-7. https://doi.org/10.1093/nar/22.24.5506
  23. Graham DI. The pathology of brain ischemia and posibilities for therapeutic intervention. Br J Anaesth. 1985;57:3-17. https://doi.org/10.1093/bja/57.1.3
  24. 辛民敎. 臨木本草學. 서울: 永林社, 1988:219-23, 249-50.
  25. Yoon HR, Yook CS. Studies on the constituents of Angelica gigas Nakai. Bull. Kyung Hee pharma. sci. 1995;23:55-71.
  26. 김성재, 송봉근, 이언정, 김형균, 김중길. 당귀 및 아위가 평활근 이완과 iNOS 발현에 미치는 영향. 大韓韓醫學會誌. 2000;20:60-7.
  27. 오하식. 당귀류 한약재가 혈관신생에 미치는 영향에 대한 비교연구. 慶熙大學校 大學院. 2001:34-5.
  28. 전연이, 박치상, 박창국. 당귀의 허혈성 뇌손상 억제작용 및 신경세포 보호효과. 대한본초학회지. 2003;18:25-35.
  29. 조정숙, 양재하, 박창국, 이희순, 김영호. 뇌졸중 치료 생약 추출물의 흥분성 신경독성 억제효과. 약학회지. 2000;44:29-35.
  30. 김수민, 조영석, 김은주, 배종만, 한준표, 이신호, 성삼경. 단삼, 도인, 당귀미 및 솔잎의 열수추출물이 지방산화에 미치는 영향. 한국식품영향과학회지. 1998;27:399-405.
  31. 李時珍. 本草鋼目. 上海:上海科學技術出版社. 1982:754-6.
  32. 김형균. 韓藥의 藥理. 서울:고려의학. 2000:90-2, 240,285-7,323.
  33. 全國韓醫科大學 本草學敎授 共編 著. 本草學. 서울:永林社. 1999:409-10,580-3.
  34. Mathews KS, McLaughlin DP, Ziabari LH, Toner CC, Street PC, Hisgrove E, Bessina EL, Stamford JA. Rapid quantification of ischemic injury and cerebroprotection in brain slices using densitometric assessment of 2,3,5-triphenylterazolium chloride staining. J Neurosci Methods. 2000;201:43-51.
  35. Fujimura M, Morita FY, Murakami K, Kawase M, Chan PH. Cytosolic redistribution of cytochrome c after transient focal cerebral ischemia in rats. J Cerb Blood Flow Metab. 1998;18:1239-47.
  36. 한국유전학회. 유전 2. 서울: 월드사이언스. 1998:147-65.
  37. Kirono T, Tamura A, Sano K. Selective vulnerability of the hippocampus to ischemia reversible and irreversible types of ischemic cell damage. Progress in Brain Res. 1985;63:39-58. https://doi.org/10.1016/S0079-6123(08)61974-3
  38. Cho Y, Bannai S : Uptake of glutamate and cystein in C6 glioma cells and in cultured astrocytes. J Neurochem., 55: 2091-2097, 1990. https://doi.org/10.1111/j.1471-4159.1990.tb05800.x
  39. Dugan LL, Sensi SL, Canzoniero LMT, Choi DW. Mitochondria production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate. J Neurosci. 1995;15:6377-88.
  40. Buttke T, Sandstrom P. Oxidative stress as a mediator of apoptosis. Immunol Today. 1994;15:7-10. https://doi.org/10.1016/0167-5699(94)90018-3