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22대 대한민국 국회 국회정보길라잡이 국회의원검색
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List of Abbreviations 8

ABSTRACT 9

I. 서론 11

II. 재료 및 방법 16

II-1. 재료 및 시약 16

II-2. 세포배양, 시약 처리 및 UVA 조사 16

II-3. 세포독성 측정 17

II-4. 활성산소종 (ROS) 생성능 측정 17

II-5. MMP-1 및 -13 효소 활성 측정 18

II-6. MMP-1 및 -13 유전자 발현 측정 18

II-7. 형질전환 19

① 형질전환을 위한 플라스미드의 제조 19

② AP-1 Luciferase활성도 측정 19

II-8. c-Jun 및 c-Fos 유전자 발현 측정 19

II-9. c-Jun 및 c-Fos 단백질 발현 측정 20

II-10. MAP kinase 활성 측정 21

II-11. 통계학적 분석 22

III. 결과 25

III-1. Apigenin과 luteolin의 세포독성 25

III-2. 활성산소종 (ROS) 형성에 대한 영향 27

III-3. MMP-1 및 -13 효소 활성 저해 29

III-4. MMP-1 및 -13 유전자 발현 억제 32

III-5. AP-1 전사인자 활성도 34

III-6. c-Jun 및 c-Fos 유전자 발현 36

III-7. c-Jun 및 c-Fos 단백질 발현 38

III-8. MAP kinases 활성 40

IV. 고찰 43

V. 참고문헌 46

List of Tables

Table 1. PCR primer sequence 23

List of Figures

Fig. 1. Model depicting solar UV irradiation damage to... 14

Fig. 2. Structure of apigenin and luteolin 15

Fig. 3. Construction of the reporter gene plasmids. Three copies... 24

Fig. 4. Effects of apigenin and luteolin on cells viability. 26

Fig. 5. Effects of apigenin and luteolin on UVA-induced cellular... 28

Fig. 6. Effect of apigenin and luteolin on the type I... 30

Fig. 7. Effect of apigenin and luteolin on the type III... 31

Fig. 8. Effects of apigenin and luteolin on UVA-induced... 33

Fig. 9. Effects of apigenin and luteolin on UVA-induced... 35

Fig. 10. Effects of apigenin and luteolin on UVA-induced... 37

Fig. 11. Effects of apigenin and luteolin on UVA-induced... 39

Fig. 12. Effects of apigenin on UVA-induced MAP kinase activity. 41

Fig. 13. Effects of luteolin on UVA-induced MAP kinase activity. 42

초록보기

Exposure of ultraviolet (UV) light for long term and repeated on the skin often induces chronic skin diseases such as skin cancer as well as photoaging, and the mechanisms of these skin damages are closely associated with up-regulation of collagenases activities. Moreover, UV-induced collagenases cause connective tissue damage and the skin to become wrinkled and aged. This study investigated the effect of apigenin and luteolin on the ultraviolet A (UVA)-induced matrix metalloproteinases (MMP-1 (collagenase-1) and MMP-13 (collagenase-3)) expression in HaCaT human keratinocytes. Treatment with apigenin and luteolin decreased the UVA-induced generation of reactive oxygen species (ROS). In addition, apigenin and luteolin significantly suppressed the UVA-induced expression of MMP-1 and -13 mRNA. Moreover, transient transfection assay using activator protein-1 (AP-1)-linked luciferase reporter gene assay revealed that apigenin and luteolin reduced UVA-induced AP-1 transcriptional activity. In this study investigated the effects of apigenin and luteolin on UVA-induced c-Jun and c-Fos expression in HaCaT cells. Apigenin and luteolin inhibited UVA-induced phosphorylation of c-Jun and expression of c-Fos. Treatment with apigenin and luteolin also inhibited ERK, JNK and p38 activation. These results suggested that apigenin and luteolin can inhibit UVA-induced MMP-1 and -13 expressions might be mediated through the AP-1 inactivation and inhibition of ROS formation in HaCaT cells. In conclusion, the results presented that apigenin and luteolin may be useful in the prevention of UVA-induced photoaging.

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