Neuroprotective Effects of Haein-tang(Hairen-tang) on Decrease of Short-term Memory and Apoptosis in Dentate Gyrus of the Gerbils with Transient Global Ischemia

해인탕이 뇌허혈 유발 모래쥐의 단기기억력 감퇴와 치상회 세포사멸에 미치는 효과

  • Park, Jung-Chul (Dept. of Oriental Rehabilitation Medicine, College of Oriental Medicine, Kyung-Won University) ;
  • Song, Yun-Kyung (Dept. of Oriental Rehabilitation Medicine, College of Oriental Medicine, Kyung-Won University) ;
  • Lim, Hyung-Ho (Dept. of Oriental Rehabilitation Medicine, College of Oriental Medicine, Kyung-Won University)
  • 박정철 (경원대학교 한의과대학 한방재활의학과교실) ;
  • 송윤경 (경원대학교 한의과대학 한방재활의학과교실) ;
  • 임형호 (경원대학교 한의과대학 한방재활의학과교실)
  • Received : 2011.03.15
  • Accepted : 2011.04.11
  • Published : 2011.04.30

Abstract

Objectives : We investigated the effect of Haein-tang(Hairen-tang) on short-term memory and apoptosis in dentate gyrus of the gerbils with transient global ischemia. Methods : For the induction of cerebral ischemia model in mice, common carotid arteries of gerbils were occluded with aneurysm clips for 5 min. One day after operation, Haein-tang(Hairen-tang) was administrated orally injected once a day for 15 consecutive days. Gerbils were randomly divided into four group(n=10 in each group): sham-operation group, ischemia-induction group, ischemia-induction and 50 mg/kg Haein-tang(Hairen-tang)-treated group, ischemia-induction and 100 mg/kg Haein-tang(Hairen-tang)-treated group, and ischemia-induction and 200 mg/kg Haein-tang(Hairen-tang)-treated group. The effect of Haein-tang(Hairen-tang) on memory function was investigated by using step-down avoidance task. Apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) staining and immunohistochemistry for caspase-3. Western blot analysis for the expressions of Bax and Bcl-2 protein was also conducted. Results : 1. Haein-tang extract significantly enhanced short-term memory in step-down avoidance task and 100 mg/kg, 200 mg/kg Haein-tang-treated group. 2. Haein-tang extract significantly suppressed TUNEL-positive cells after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg Haein-tang-treated group. 3. Haein-tang extract significantly increased caspase-3 positive cells in the hippocampal dentate gyrus after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg Haein-tang-treated group. 4. Haein-tang extract significantly decreased Bax protein expressions in the hippocampus after transient global ischemia and 100 mg/kg, 200 mg/kg, Haein-tang-treated group. Haein-tang extract significantly increased Bcl-2 protein expressions in the hippocampal dentate gyrus after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg, Haein-tang-treated group. Haein-tang extract significantly decreased Ratio of Bax protein to Bcl-2 protein in the hippocampus after transient global ischemia and 100 mg/kg, 200 mg/kg Haein-tang-treated group. Conclusions : While Haein-tang(Hairen-tang) treatment improved short-term memory by suppressing on ischemia-induction apoptosis. In the present study, Haein-tang(Hairen-tang) shows protective effect on transient global ischemia.

Acknowledgement

Supported by : 경원대학교

References

  1. Biebl M, Cooper CM, Winkler J, Kuhn HG. Analysis of neurogenesis and programmed cell death reveals a self-renewing capacity in the adult rat brain. Neurogenesis Letters. 2000;291(1):17-20.
  2. Eriksson PS, Perfilieva E, Bjork-Eriksson T, Alborn AM, Nordborg C, Peterson DA, Gage FH. Neurogenesis in the adult human hippocampus. Nature Medicine. 1998:4(11):1313-7. https://doi.org/10.1038/3305
  3. Sim YJ, Kim H, Kim JY, Yoon SJ, Kim SS, Chang HK, Lee TH, Lee HH, Shin Me, Shin MS, Kim CJ. Long-term treadmill exercise overcomes ischemia-induced apoptotic neuronal cell death in gerbils. Physiol Behav. 2005;84:733-8. https://doi.org/10.1016/j.physbeh.2005.02.019
  4. Ko IG, Shin MS, Kim BK, Kim SE, Sung YH, Kim TS, Shin Mc, Cho HJ, Kim SC, Kim SH, Kim KH, Shin DH, Kim CJ. Tadalafil improves short-term memory by suppressing ischemia-induced apoptosis of hippocampal neuronal cells in gerbils. Pharmacol Biochem Behav. 2009;91:629-35. https://doi.org/10.1016/j.pbb.2008.10.009
  5. Dirnagl U, Iadecola C, Moskowitz MA. Pathpbiology of ischemicc stroke: an integrated view. Trends in Neurosciences. 1999;22(9):391-7. https://doi.org/10.1016/S0166-2236(99)01401-0
  6. Leker RR, Shohami E. Cerebral ischemia and trauma-different etiologies yet similar mechanisms: neuroprotective opportunities. Brain Research. 2002;39(1):55-73. https://doi.org/10.1016/S0165-0173(02)00157-1
  7. Love S, Barber R, Wilcock GK. Neuronal death in brain infarcts in man. Neuropathology and Applied Neurobiology. 2000;26(1):55-66. https://doi.org/10.1046/j.1365-2990.2000.00218.x
  8. 한태륜, 박문식. 재활의학. 서울:군자출판사. 2008:123-7.
  9. Lee MH, Kim H, Kim SS, Lee TH, Lim BV, Chang HK, Jang MH, Shin Mc, Shin MS, Kim CJ. Treadmill exercies suppresses ischemia-induced increment in apoptosis and cell proliferation in hippocampal dentate gyrus of gerbils. Life Sci. 2003;73:2455-65. https://doi.org/10.1016/S0024-3205(03)00655-6
  10. 우주영, 김종우, 황의완, 김현택, 박순권. 調胃升淸湯이 흰쥐의 방사형 미로 학습과 기억에 미치는 영향. 동의신경정신과학회지. 1997;8(1):12-8.
  11. 조윤숙, 황의완, 김현택, 박순권. 荊防地黃湯이 Alzheimer's disease 모델백서의 학습과 기억에 미치는 영향. 동의신경정신과학회지. 1998;9(1):35-44.
  12. 조윤숙, 황의완, 김현택, 박순권. 荊防地黃湯이 Alzheimer's disease 모델백서의 학습과 기억에 미치는 영향. 동의신경정신과학회지. 1998;9(1):35-44.
  13. 정성욱, 장규태, 김장현. 일시적 국소 뇌허혈 휜쥐모델에서 寶丹祛風至의 뇌손상 및 인지기능 보호효과. 대한한의학회지. 2005;26(2):52-62.
  14. 이준환, 송미연, 이종수, 김성수, 신현대, 정석희. Mongolian gerbil의 뇌허혈에 대한 현삼의 신경보호효과. 한방재활의학과학회지. 2008;18(4):1-11.
  15. 김기현, 민상연, 김장현. 육미지황탕이 국소뇌허혈유발 기억장애 모델 흰쥐에 미치는 영향. 대한한의학회지. 2009;30(2):1-16.
  16. Jang MH, Shin MC, Lee TH, Lim BV, Shin MS, Min BI, Kim H, Cho S, Kim EH, Kim CJ. Acupuncture suppresses ischemia-induced increase in c-Fos expression and apoptosis in the hippocampal CA1 region in gerbils. Neurosci Lett. 2003;347:5-8. https://doi.org/10.1016/S0304-3940(03)00512-3
  17. Deng HZ, Sun SY, Li SY. An experimental observation on the analgesic and antipyretic effevts and toxicity of Acanthopanax giraldii Harms. Zhongguo Zhong Yao Za Zhi. 1994;19(1):38-40, 63.
  18. 김상태, 한용남, 손연경, 장형석, 김수장, 신준식. 구척으로부터 신경재생 효능 성분 분리. 약학회지. 2002;46:398-404.
  19. 박원상, 김은석, 신정원, 구척이 흰쥐의 척수압박에 의한 신경세포 손산에 미치는 영향. 한방재활의학과학회지. 2010;20(2):1-15.
  20. 윤덕영, 박성하, 이종수. 杜沖이 근육위축 흰쥐의 후지 근섬유형 및 MyoD 발현에 미치는 영향. 한방재활의학과학회지. 2008;18(1):47-63.
  21. 한방재활의학과학회. 한방재활의학. 서울:군자출판사. 2006:133-87.
  22. Lee YC, Kwon TH, Ok IS, Sen CW, Kim YJ, Roh SS, Seo YB. The experimental studies on the immunomodulational effects of Lonicerae Caulis et Folium. Kor Journal herbology. 2005;20:141-9.
  23. Lapchak PA The phenylpropanoid micronutrient chlorogenic acid improves chinical rating scores in rabbits following multiple infarct ischemic strokes: synergism with tissue plasminogen activator. Exp Neurol. 2007;205:407-13. https://doi.org/10.1016/j.expneurol.2007.02.017
  24. 김종철, 이은용, 김호현, 오가피 약침이 알코올 중독 휜쥐의 치상회에서 신경세포생성 및 NOS 발현에 미치는 영향. 대한경락경혈학회지. 2003;20(2):67-76.
  25. 김상태, 김정도, 김영균. 구척(狗脊) 메탄올추출액이 신경세포의 재생 및 회복효과에 미치는 영향. 생약학회지. 2004;35(2):105-9.
  26. 김광진, 김창주, 김형진, 박사훈. 인체생리학. 서울:정문각. 1998:425-6.
  27. 전국한의과대학 심계내과학 교실. 심계내과학. 서울:군자출판사. 2006:375-8.
  28. Cramer SC, Chopp M. Recovery recapitulates ontogeny. Trends Neurosci. 2000;23:265-71. https://doi.org/10.1016/S0166-2236(00)01562-9
  29. Sims NR, Anderson MF. Mitochondrial contributions to tissue damage in stroke. Neurochem Int. 2002;40:511-26. https://doi.org/10.1016/S0197-0186(01)00122-X
  30. Hengartner MO. The biochemistry of apoptosis. Nature. 2002;407:770-7.
  31. Squire LR, Zola SM. Ischemic brain damage and memory impairment: a commentary. Hippocampus. 1996;6:546-52. https://doi.org/10.1002/(SICI)1098-1063(1996)6:5<546::AID-HIPO7>3.0.CO;2-G
  32. 한의과대학 본초학 편찬위원회. 본초학. 서울:영림사. 2004:165-6, 267-8, 312-3, 328-9, 469-70, 605-7, 611-2.
  33. 김의종, 김연섭. 양격산화탕(凉膈散火湯)이 당뇨흰쥐의 전뇌허혈에 미치는 영향. 동의생리병리학회지. 2008;22(2):321-7.
  34. 이환성, 박성준, 정광식, 손영주, 정혁상, 박동일, 손낙원. 양격산화탕(凉膈散火湯)이뇌해마조직배양의 허혈손상에 따른 신경세포손상에 미치는 영향. 대한한방내과학회지. 2008;29(1):231-42.
  35. 윤영내, 최찬헌, 백진웅, 김형우, 윤내한, 김경윤, 남기원, 김계엽, 정현우. 오갈피나무 뿌리 50% 에탄올 추출물이 항허혈에 미치는 실험적 효과. 동의생리병리학회지. 2007;21(4):891-7.
  36. 박세홍, 이광로, 배선준, 정상수, 강세영, 이상관, 이성근, 윤지원, 성강경. 大造丸이 大腦神經細胞의 虛血性 損傷에 미치는 영향. 동의생리병리학회지. 2003;17(6):150-8.
  37. 김봉상, 이선우, 문병순. 허혈 상태의 PC12 세포에 대한 대보원전(大補元煎)의 신경보호효과. 동의생리병리학회지. 2007;21(1):117-25.
  38. 조은희, 권정남, 김영균. 혈부축어탕이 흰쥐의 국소대뇌허혈에 미치는 신경보호 효과. 대한한의학회지. 2006;27(2):70-85.
  39. Niwa M, Hara A, Iwai T, Wang S, Hotta K, Mori H, Uematsu T. Caspase activation as an apoptotic evidence in the gerbil hippocampal CA1 pyramidal cells following transient forebrain ischemia. Neurosci Lett. 2001;300:103-6. https://doi.org/10.1016/S0304-3940(01)01559-2
  40. Kim YM, Park KN, Choi SP, Hong TY, Kim SK. The effects of mild hypothermia on the expression of the apoptosis-related proteins following transient global ishcemia in gerbil hippocampus. Korean J Crit Care Med. 2007;22:30-41.
  41. Lee JH, Kwon O, Kwon JY. The effects of desflurane on delayed neuronal injury after transient forebrain ischemia in the rat. Korean J Anesthesiol. 2009;57:195-202. https://doi.org/10.4097/kjae.2009.57.2.195
  42. He H, Lam M, McCormick TS, Distelhorst avo Maintenance of calcium homeostasis in the endoplasmic reticulum by Bcl-2. J Cell Bio. 1997;138:1219-22. https://doi.org/10.1083/jcb.138.6.1219
  43. Min HK, Kang HC, Lee HS, Kim SY, Sohn Y, Jung HS, Sohn NW, Kim YB. Neuroprotecticve Effect of Sophora Subprostrata on transiet global ischemia in gerbil. Kor J Herbology. 2008;23:1-9.
  44. Hockenbery D, Nunez G, Milliman C, MLiliJmn C, Schreiber RD, Korsrreyer SJ. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990;348:334-6. https://doi.org/10.1038/348334a0
  45. Martinou JC, Dubois-Dauphin M, Staple JK, Rodriguez I, Frankowski H, Missotten M, Albertini P, Talabot D, Catsicas S, Pietra C, Huerte J. Overexpression of Bcl-2 in transgenic mice protects neurons from naturally occurring cell death and experimental ischemia. Neuron. 1994;13:1017-30. https://doi.org/10.1016/0896-6273(94)90266-6