Title Page
Contents
국문요약 9
Abstact 11
I. Introduction 13
II. Materials and Methods 15
1. Materials 15
2. Cell culture 15
3. Cell viability assay 16
4. RNA extraction and real time RT-PCR 17
5. Western blot analysis 19
6. TEER (Transepithelial Electrical Resistance) assay 22
7. Statistical processing 23
III. Results and Discussion 23
8. Brazilin is structure and features 23
9. Viability of TNF-α and brazilin in HaCaT keratinocyte cells 25
10. Inhibitory effects of brazilin on the mRNA expression levels pro-inflammatory cytokines in TNF-α induced HaCaT cells 27
11. Brazilin suppressed CCL20 mRNA expression of chemokine on TNF-α induced HaCaT cells 30
12. Brazilin down-regulated phosphorylation of I-κB in TNF-a stimulated HaCaT cells 32
13. Brazilin down-regulated phosphorylation of Akt in TNF-a stimulated HaCaT cells 34
14. Decreased protein level expression of phosphorylation of ERK by brazilin 36
15. Decreased protein level expression of phosphorylation of JNK by brazilin 38
16. Decreased protein level expression of phosphorylation of p38 by brazilin 40
17. Brazilin suppressed phosphorylation of STAT3 in TNF-a stimulated HaCaT cells 42
18. Enhancement effects of brazilin on the skin barrier in TNF-a stimulated HaCaT cells 44
IV. Conclusion 47
V. References 51
Appendix 57
Appendix 1. Appl Biol Chem 62, 46 (2019) doi:10.1186/s13765-019-0455-z 57
Appendix 2. 2019년 5월 17일 제 44차 대한화장품학회 춘계학술대회 58
List of Tables
Table 1. Gene name and assay ID number in real-time PCR analysis 18
Table 2. Antibody list used in Western blot analysiss 21
Fig. 1. Structure of Brazilin and form of Heat wood (蘇木). brazilin is... 24
Fig. 2. Viability of HaCaT cells after treatment with TNF-α and... 26
Fig. 3. Analysis of gene expression of the proinflammatory cytokines... 29
Fig. 4. Analysis of gene expression of chemokine (C-C motif) ligand... 31
Fig. 5. Brazilin suppressed phosphorylation of IκB and Akt, not NF-κ... 33
Fig. 6. Brazilin suppressed phosphorylation of Akt in TNF-a... 35
Fig. 7. Brazilin down-regulated phosphorylation of ERK protein related... 37
Fig. 8. Brazilin down-regulated phosphorylation of JNK protein related... 39
Fig. 9. Changes in protain expression levels of p38 analyzed by... 41
Fig. 10. Brazilin suppressed phosphorylation of STAT3 in TNF-a... 43
Fig. 11. The TEER value was measured to verify the effect of... 46