Ischemic stroke is caused by a blockage of the cerebral artery, which leads to a severe neurological disorder. Chlorogenic acid is a phenolic acid found mainly in plants such as coffee beans, eggplants, and carrots. It exerts a neuroprotective effect against cerebral ischemic damage. Bcl-2 family protein is a representative apoptosis regulatory protein. Bcl-2 and Bcl-xL act as apoptosis inhibitors, while Bax and Bad act as apoptosis inducers.The interaction of Bcl-2 family protein plays an important role in determining cell fate. The aim of this study was to investigate whether chlorogenic acid modulates the interaction of Bcl-2 family proteins during ischemic injury. Middle cerebral artery occlusion (MCAO) surgery was performed to induce cerebral ischemia. Chlorogenic acid (30 mg/kg) or phosphate buffered saline was intraperitoneally injected to adult male rats 2 h after MCAO surgery. Neurobehavioral tests were performed to confirm the neuroprotective effect of chlorogenic acid 24 h after MCAO injury, and immunoprecipitation analysis was performed to investigate the interaction of Bcl-2 family protein. MCAO damage showed signs of severe neurological disorders, while chlorogenic acid improved these disorders. Results of immunoprecipitation analysis were as follows. Interaction between Bax and Bcl-2 or Bcl-xL was decreased in MCAO injury, chlorogenic acid prevents these decreases. In contrast to Bax, Interaction between Bad and Bcl-2 or Bcl-xL was increased in MCAO injury, chlorogenic acid prevents these increases. Furthermore, chlorogenic acid attenuated MCAO-induced increase of capase-9. In conclusion, our findings demonstrate that chlorogenic acid exerts a neuroprotective effect against cerebral ischemic injury by modulating interaction of Bcl-2 family proteins.

서 론
재료 및 방법
    실험동물 준비
    중간대뇌동맥폐쇄술 (middle cerebral artery occlusionsurgery)
    신경학적 결손 점수 검사 (neurological deficit scoringtest)
    코너 검사 (corner test)
    면역침강법 (immunoprecipitation analysis)
    통계분석
결 과
    MCAO 손상시 클로로겐산의 신경보호 효과
    MCAO 손상시 클로로겐산 투여에 의한 Bcl-2와 Bax,Bcl-xL과 Bax의 상호작용의 조절
    MCAO 손상시 클로로겐산 투여에 의한 Bcl-2와 Bad,Bcl-xL과 Bad의 상호작용의 조절
    MCAO 손상시 클로로겐산 투여에 의한 caspase-9발현의 조절
고 찰
감사의 글
REFERENCES
Supplement 1

• Ju-Bin Kang(Department of Veterinary Medicine, College of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea)
• Phil-Ok Koh(Department of Veterinary Medicine, College of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea) Corresponding author