Title Page

Contents

Abstract 9

Ⅰ. INTRODUCTION 11

1. Skin structure 11

A. Composition of the epidermis 11

B. Cornified envelope (CE) formation explanation 11

2. Aryl hydrocarbon receptor (AhR) 12

A. Signaling pathway of AhR 12

B. Barrier enhancement by AhR 13

C. Skin inflammation and AhR 15

D. Therapeutic potential of AhR ligands for inflammatory skin diseases 15

E. Antioxidant activity of AhR 16

3. Cannabidiol (CBD) 16

A. Biological activity of CBD 16

B. Receptors for CBD 17

C. Function CBD in the skin 18

4. Research purpose 19

Ⅱ. MATERIALS AND METHODS 20

1. Reagent and antibodies 20

2. Cell culture 20

3. AhR luciferase assay 21

4. MTT assay 22

5. Cell apoptosis assay 22

6. Reactive oxygen species (ROS) assay 23

7. Immunocytochemistry (ICC) 24

8. Western blot 24

9. Total RNA extraction and quantitative real-time PCR 25

10. Statistical analysis 26

Ⅲ. RESULTS 29

1. Cannabidiol (CBD) regulates the activation of arylhydrocarbon receptor (AhR) 29

2. CBD-induced AhR nuclear translocation and increased AhR target genes 33

3. Cannabinoid receptors did not mediate CBD-induced CYP1A1 expression 36

4. Knockdown of AhR inhibited CBD-induced CYP1A1 and AhRR expression 39

5. CBD upregulates skin barrier proteins via the AhR-OVOL1 axis 42

6. CBD scavenges reactive oxygen species by regulating antioxidant enzymes via the AhR pathway 45

7. Expected mechanism of action of CBD in keratinocytes. 48

Ⅳ. DISCUSSION 51

Ⅴ. CONCLUSION 53

REFERENCES 54

ABSTRACT IN KOREAN 62

Table 1. Information of primary antibodies 27

Table 2. Information of primers 28

Figure 1. Cannabidiol (CBD) regulates the activation of arylhydrocarbon receptor (AhR) 31

Figure 2. CBD-induced AhR nuclear translocation and increased AhR target genes 34

Figure 3. Cannabinoid receptors did not mediate CBD-induced CYP1A1 expression 37

Figure 4. Knockdown of AhR inhibited CBD-induced CYP1A1 and AhRR expression 40

Figure 5. CBD upregulates skin barrier proteins via the AhR-OVOL1 axis 43

Figure 6. CBD scavenges reactive oxygen species by regulating antioxidant enzymes via AhR pathway 46

Figure 7. Expected mechanism of action of CBD in keratinocytes. 49