Title Page
Contents
ABBREVIATIONS 9
국문요약 11
I. INTRODUCTION 13
II. MATERIALS AND METHODS 19
1. Schisandra chinensis seed extracts 19
2. Cell culture and treatments 19
3. Cell viability test 19
4. Reverse transcriptase-polymerase chain reaction (RT-PCR) 20
5. Western blot analysis 22
6. Nitric oxidase (Griess Reagent assay) 24
7. Determination of AChE Activity 24
8. Statistical analysis 25
III. RESULTS 26
1. Cytotoxicity of SCSE on pheochromocytoma cell line PC12 cells 26
2. Effects of SCSE on LPS-induced cell death 28
3. Effects of SCSE on TLR4-related proteins in LPS-treated PC12 cells 30
4. Effect of SCSE on MAPK-related protein activation in LPS-treated PC12 cells 32
5. Effects of SCSE on pro-inflammatory cytokine in LPS-treated PC12 cells 34
6. Effects of SCSE on NO Production and iNOS/COX-2 Expression in LPS-Stimulated PC12 cells 37
7. Measurement of AchE activity 40
IV. DISCUSSION 42
V. REFERENCE 46
Abstract 53
Table 1. List of primer sequences and PCR conditions for RT-PCR 21
Table 2. Antibodies used in the present study 23
Figure 1. Schematic diagram showing the inflammatory signaling pathway of LPS-induced TLR4 16
Figure 2. Structure of the cholinergic nervous system. 18
Figure 3. Assessment of cytotoxicity of SCSE alone in PC12 cells 27
Figure 4. Effects of LPS-induced cell viability on SCSE in PC12 cells 29
Figure 5. Effects of SCSE on pro-inflammatory mediators in LPS-treated PC12 cells 31
Figure 6. Effects of SCSE on the phosphorylation of MAPK-related protein in LPS-induced PC12 cells 33
Figure 7. Effects of SCSE on the mRNA expression levels of pro-inflammatory cytokine 35
Figure 8. Effects of SCSE on the expression levels of pro-inflammatory cytokine proteins 36
Figure 9. Effects of LPS-induced nitric oxide level on SCSE in PC12 cells 38
Figure 10. Effects of SCSE on the expression levels of iNOS and COX-2 proteins 39
Figure 11. Inhibitory effect of SCSE on AchE activity 41