Title Page
Contents
ABSTRACT 9
Ⅰ. Introduction 10
Ⅱ. Material and Methods 12
2.1. C. verum extract preparation 12
2.2. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) 12
2.3. Cell culture 13
2.4. Cell viability assay 13
2.5. RNA isolation and RT-qPCR 13
2.6. Intracellular ROS determination 14
2.7. Immunoblot analyses 14
2.8. Immunofluorescence staining 15
2.9. Statistical analysis 16
Ⅲ. Results 17
3.1. Effects on the cell viability and morphology of THP-1 cells 17
3.2. Effects on PMA-stimulated PKC activation in THP-1 cells 19
3.3. Effects on ROS levels in PMA-stimulated THP-1 cells 22
3.4. Effects of PKCδ inhibition on PMA-stimulated ROS levels and NOX2 gene expression in PMA stimulated THP-1 cells 24
3.5. Effects on mRNA levels of inflammatory mediators in PMA-stimulated THP-1 cells 25
3.6. Effects on the MAPK pathway and transcription factors in PMA-stimulated THP-1 cells 27
3.7. Constituents of methanol extract of C. verum and their activities 29
Ⅳ. Discussion 34
References 38
Abstract (in Korean) 41
Table 1. Primer sequences. 14
Table 2. Non-targeted analysis of Cinnamomum verum extract using UPLC-QTOF/MS. 31
Figure 1. Cytotoxic effects of C. verum on THP-1 cells. (A) THP-1 cells were treated with C. verum (20 and 40 µg/mL) for 24 h. (B) THP-1 cells were co-treated... 18
Figure 2. Effects of C. verum on PKC activity in PMA-treated THP-1 cells. (A) PKC substrate phosphorylation was evaluated using immunoblotting. THP-1 cells were... 20
Figure 3. Effects of C. verum on the ROS levels in PMA-treated THP-1 cells. (A) ROS levels were evaluated using DCF-DA staining. THP-1 cells were stained by... 23
Figure 4. Effects of PKCδ inhibitor on PMA-stimulated ROS levels and NOX2 gene expression in THP-1 cells. (A) THP-1 cells were stained by DCF-DA and pre-treated... 24
Figure 5. Effects on IL-1β, IL-8, CCL5, and COX-2 expression in PMA-treated THP-1 cells. (A) THP-1 cells were pre-treated with C. verum (20 and 40 µg/mL) for 1 h,... 26
Figure 6. Effects of C. verum on the MAPK phosphorylation and nuclear translocation of transcription factors. (A) Phosphorylation levels of ERK, JNK, and... 28
Figure 7. The UPLC-QTOF/MS chromatograms of the MeOH extract of C. verum. (-) ESI base peak chromatograms (A) and (+) ESI base peak chromatograms (B) of C.... 30
Figure 8. Effects of active compound of C. verum on PKCδ/NOX2/ROS and pro-inflammatory genes in PMA stimulated THP-1 cells. THP-1 cells were pre-treated... 33
Figure 9. Schematic diagram of the effect of C. verum on the PMA-treated THP-1 cells. Schematic diagram of the effect of C. verum on the PMA-treated THP-1 cells. C. verum... 37