Functional Recovery and Cognitive Behavior Enhancing Effects of Chong-Myung-Tang in the Ischemic Stroke Rat and its Neuroprotective Properties : 총명탕이 허혈성 뇌졸중 유발 흰쥐의 기능적 회복 및 인지 행동 촉진에 미치는 영향과 신경보호 특성에 관한 연구

다국어 초록 (Multilingual Abstract)

PURPOSE: Chong-Myung-Tang (CMT) consisted of Polygala tenuifolia Willdenow, Acorus Gramineus Solander and Poria cocos Wolf is a famous polyherbal formula of Korean Medicine used for the treatment of learning and memory improvement. The object of this study was to investigate the effects of CMT on sensorimotor function, cognitive behavior, and to observe neuroprotective effects in ischemic stroke rat.
METHODS: Three different dosages of CMT aqueous extracts, 500, 250 and 125 / were orally administered, dissolved in distilled water, once a day for 28 continuous days from 24h after end of the permanent occlusion of the middle cerebral artery (pMCAO). The sensorimotor function and cognitive behavior recovery effects of CMT in the ischemic stroke stroke induced pMCAO were observed by evaluating sensorimotor function and cognitive behavior tests. Furthermore, in order to observe neuroprotective and antioxidant effects of CMT in cerebral cortex after pMCAO, histopathological changes, the cerebral cortex malondialdehyde (MDA) contents for lipid peroxidation, glutathione (GSH) contents, superoxide dismutase (SOD), catalase (CAT) activities, cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP) as markers of programmed cell death (PCD) in cerebral ischemia, nitrotyrosine as marker of inducible nitric oxide synthase (iNOS) related oxidative stresses and 4- hydroxynonenal (4-HNE) as marker of lipid peroxidation were investigated. The results were compared with those of donepezil 10 / as reference drug.
RESULTS: Ischemic damages induced pMCAO around peri-infarct/defect cerebral cortex, disorders of sensorimotor function and cognitive behavior, destroys of antioxidant defense systems, and execution of PCD were markedly and dose-dependently inhibited by treatment of CMT 500, 250 and 125 / , respectively.
CONCLUSION: The results suggest that CMT could accelerate the recovery of sensorimotor function and enhance cognitive behavior in the ischemic stroke rat. Moreover, ischemic brain damages could be reduced by the antioxidant and neuroprotective effects of CMT.

국문 초록 (Abstract)

실험목적: 총명탕은 원지, 석창포 및 복신의 3종의 약재로 구성되어 있는 처방으로 건망 및 치매 등의 병증에 활용되고 있는 처방이다. 본 연구는 총명탕이 영구적인 중대뇌동맥폐쇄 (permanent occlusion of the middle cerebral artery, pMCAO)로 유발된 허혈성 뇌졸중 흰쥐의 기능적 회복과 인지 행동 촉진에 미치는 영향을 알아보았다.
실험방법: 멸균증류수에 용해시킨 총명탕 열수 추출물을 500, 250 및 100 ㎎/㎏의 용량으로 각각 pMCAO 유발 24시간 후부터 매일 1회씩 28일간 경구투여하고, 감각운동 기능 및 인지 행동의 변화를 관찰하였다. 그리고 infarct/defect 주변부위의 대뇌 위축 및 변성 신경세포의 수의 변화를 조직학적으로 관찰하였고, infarct/defect 주변부 대뇌 피질의 항산화 방어 시스템 (malondialdehyde (MDA) 및 glutathione (GSH) 함량, superoxide dismutase (SOD) 및 catalase (CAT) 활성의 변화)을 관찰하였다. 또한 세포예정사 (programmed cell death, PCD) 지표인 cleaved caspase-3 및 cleaved poly (ADP-ribose) polymerase (PARP), 지질과산화 지표인 nitrotyrosine, inducible nitric oxide synthase (iNOS) 관련 oxidative stress 지표인 4-hydroxynonenal (4-HNE)의 면역반응 신경세포의 수적 변화를 면역조직화학적 방법으로 각각 관찰하였다.
결과: pMCAO에 의한 감각운동 기능 및 인지 행동 장애, infarct/defect 주변부위 대뇌 피질 신경원의 변성, 항산화 방어시스템의 손상 및 PCD가 총명탕 500, 250 및 125 ㎎/㎏ 투여에 의해 투여 용량 의존적으로 유의성 있게 현저히 억제되었다.
결론: 이상의 결과에서, 총명탕은 허혈성 뇌손상에 따른 감각운동 기능 및 인지 행동 장애에 매우 유효한 효과를 나타내고, 항산화 방어 시스템의 활성 및 신경세포 보호 효과를 가지고 있는 것으로 관찰된다.

목차 (Table of Contents)

• I. INTRODUCTION 1
• II. MATERIALSANDMETHODS 4
• 1. Animals and husbandry 4
• 2. Preparations and administration of test materials 7
• 3. Permanent occlusion of the middle cerebral artery (pMCAO) 10
• 4. Body weight measurements 10
• 5. Assessment of the sensorimotor function 11
• 6. Assessment of the cognitive behavior 13
• 7. Analysis of the infarct/defect volumes 13
• 8. Histopathological procedure 14
• 9. Assessment of the antioxidant defense systems 15
• 10. Immunohistochemistry 16
• 11. Statistical analysis 19
• III. RESULTS 20
• 1. Changes on the body weight 20
• 2. Effects on the sensorimotor function 23
• 1) Forelimb placing scores 23
• 2) Hindlimb placing scores 25
• 3) Elevated body swing test (EBST) 27
• 3. Effects on the cognitive behavior 30
• 4. Changes on the infarct/defect volumes 33
• 5. Effects on the peri-infarct/defect cerebral cortex 37
• 1) Changes on the atrophic % of ipsilateral cerebral cortex 37
• 2) Changes on the percent cell numbers of degenerative cerebral 37
• 6. Effects on the peri-infarct/defect cerebral antioxidant defense 41
• 1) Changes on the peri-infarct/defect cerebral MDA levels 41
• 2) Changes on the peri-infarct/defect cerebral GSH contents 41
• 3) Changes on the peri-infarct/defect cerebral CAT activities 41
• 4) Changes on the peri-infarct/defect cerebral SOD activities 42
• 7. Effects on the cerebral cortex cleaved caspase-3 and cleaved 44
• 1) Changes on the cleaved caspase-3-immunolabeled cerebral 44
• 2) Changes on the cleaved PARP-immunolabeled cerebral neurons 44
• 8. Effects on the cerebral cortex nitrotyrosine and 4-HNE- 48
• 1) Changes on the nitrotyrosine-immunolabeled cerebral neurons 48
• 2) Changes on the 4-HNE-positive cerebral neurons 48
• IV. DISCUSSION 52
• V. CONCLUSION 60
• REFERENCES 61
• (초록) 70
• (Abstract) 71