Title Page
Abstract
Contents
Ⅰ. Introduction 11
Ⅱ. Materials and Methods 14
1. Chemicals and reagents 14
2. Preparation of ethanol extract 14
2-1. Extract by extraction method 15
2-2. Extract by composition of solvent 15
3. High performed liquid chromatography (HPLC) analysis 16
4. Method validation 18
5. ROS scavenging assay 19
6. Cell culture and treatment 20
7. Measurement of cell viability 21
8. Analysis of intracellular ROS levels 22
9. Gene expression analysis 23
10. Western blot 24
11. Statistical analysis 25
Ⅲ. Results and Discussion 26
1. Method validation 26
1-1. Specificity 26
1-2. Linearity 28
1-3. LOD and LOQ 30
1-4. Precision and accuracy 32
1-5. Quantification analysis 34
2. Effects of Schisandra chinensis on ROS scavenging activity 38
3. Cell viability effects of SCE and its active components on RAW 264.7 cell 40
4. Effects of Shisandra chinensis on intracellular ROS levels 42
5. Effects of SCE and its active components on gene expression 44
6. Effects of SCE and its active components on inflammasome 46
Ⅳ. Conclusions 48
References 51
개요 58
Table 1. The conditions of liquid chromatography. 17
Table 2. Calibration curves, LOQs and LODs for lignans 31
Table 3. Inter- and intra-day precision and accuracy of lignans 33
Table 4. Contents of lignans in Schisandra chinensis ethanol extracts by extraction method 35
Table 5. Contents of lignans in Schisandra chinensis aqueous ethanol extracts by composition of solvent 36
Table 6. Contents of lignans in Schisandra chinensis 60% aqueous ethanol under reflux method extracts 37
Figure 1. HPLC chromatograms of 5 kinds of lignans (A) and Schisandra chinensis extract (B). 27
Figure 2. Calibration curve of 5 kinds of lignan standards. 29
Figure 3. ROS scavenging activity of SCE (A), lignans from SCE (B), GN (C), Sch C (D). The reactive oxygen species (ROS) scavenging activity was... 39
Figure 4. Changes in the viability of RAW 264.7 cells after incubation of SCE, GN, and Sch C. Cell viability of macrophages after exposure to SCE (A), GN (B), and Sch C (C) for 24h (A). One-factor ANOVA was used to... 41
Figure 5. Intracellular ROS level inhibited by SCE (A), GN (B), and Sch C (C). Cell lysates were analyzed to determine whether SCE and major components inhibited LPS-induced ROS levels (n=8). One-factor ANOVA was used to... 43
Figure 6. The effect of SCE, GN, and Sch C on IL-1β, IL-6, and TNFα production in RAW 264.7 cells by concentration. Cell lysates were analyzed for pro-inflammatory cytokine levels by Q-PCR (n=6). SCE and GN reduced... 45
Figure 7. The effect of GN and Sch C on NLRP3 inflammasome inactivation in LPS-induced RAW 264.7 cells. Cell lysates were analyzed for levels of NLRP3... 47