표제지
목차
국문초록 9
I. 서론 11
II. 재료 및 방법 13
1. 실험동물 13
2. HMD 및 HMDP 제조방법 13
3. 실험방법 14
(1) High-Performance Liquid Chromatography(HPLC)을 이용한 HMD 구성성분 분석 14
(2) 세포배양 15
(3) 세포 생존율 측정 15
(4) 실험동물 군 분리 16
(5) 약물투여 17
(6) HMD 약침요법 및 혈위 18
(7) Morris water maze 19
(8) Y-maze 20
(9) 해마조직 제작 20
(10) H&E staining 21
(11) Cresyl violet staining 21
(12) Reverse Transcription Polymerase Chain Reaction (RT-PCR) 22
(13) Quantitative-PCR (qPCR) 24
4. 통계 처리 25
III. 실험결과 26
1. HMD 구성 성분에 대한 HPLC 분석 26
2. 신경세포 사멸 보호 효과 28
3. 동물 행동학적 변화 30
(1) Morris water maze로 측정한 acquisition training 30
(2) Morris water maze로 측정한 spatial memory 32
(3) Y-maze로 측정한 working memory 35
4. Hippocampus staining 변화 37
(1) 세포핵막 염색 37
(2) 신경세포 밀도 39
5. RT-PCR analysis 변화 41
6. qPCR analysis 변화 43
IV. 고찰 45
V. 결론 52
참고문헌 54
Abstract 64
Table 1. Composition of Hayeong-Mansu-Dan(HMD) 14
Table 2. Experiment animal group division 16
Table 3. Sequence of the primers used in RT-PCR analysis 23
Table 4. Sequence of the primers used in qPCR analysis 25
Table 5. Cell viability for hippocampus HT-22 cell lines after glutamate... 28
Table 6. Times to reach the platform during the acquisition training 30
Table 7. Time spent in each quadrant during the spatial memory test 33
Table 8. Spatial memory, number of time, distance, speed and during... 33
Table 9. Distance, number of entery, alternation during working... 35
Table 10. mRNA expression of BDNF, CREB, and mAChR using... 43
Fig. 1. Animal experimental design. 17
Fig. 2. BL15 and ST36 position. 18
Fig. 3. HPLC chromatogram for 5 standards. 27
Fig. 4. Effect of HT-22 cell lines death on glutamate induction by... 29
Fig. 5. The effect of HMD and HMDP on the time platform for 4 days... 31
Fig. 6. The effect of HMD and HMDP on the spatial memory after... 34
Fig. 7. The effect of HMD and HMDP on the working memory after... 36
Fig. 8. Comparison of cell nuclear membrane in hippocampus CA1, CA3, DG... 38
Fig. 9. The density of nerve cells in the region CA1, CA3 and DG of... 40
Fig. 10. Effect of HMD and HMDP on the expression of Bcl-2, Bax,... 42
Fig. 11. Effect of HMD and HMDP on the expression of BDNF,... 44
Fig. 12. Cognitive function improvement effect of HMD and HMDP. 51