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Ⅰ. Introduction 6
Ⅱ. Materials and methods 8
1. Materials 8
1) Reagents and apparatus 8
2) Experimental animals 9
3) Drugs 10
2. Procedure 12
1) Drug preparation 12
2) Drug administration into the NC/Nga atopy dermatitis mice 12
3) Clinical skin severity 13
4) Cytokine assay in the serum 13
5) Fluorescence Activated Cell Sorting analysis 13
6) Reverse Transcription Polymerase Chain Reaction 14
7) IL-6 and IL-13 quantitation in the skin tissues and cell surface molecule analysis by FACS 16
8) Histopathological examination 17
9) Lymphokine assay 17
10) Statistical analysis 17
Ⅲ. Results 19
1. Determination of AD severity in NC/Nga mice after drug administration 19
2. Effects of TG and ET on Serum IgE and IL-6 levels in AD mice 21
3. Effects of TG and ET on Serum IL-4 and IL-5 levels in NC/Nga mice 23
4. Effects of TG and ET on Serum IgE and IL-13 and IFN-γ levels in NC/Nga mice 24
5. Effects of TG and ET on spleen weight of NC/Nga mice 25
6. Effect of TG and ET on serum IgM in NC/Nga mice 26
7. Effects of TG and ET on serum IgG1 and IgG2a in NC/Nga mice 27
8. FACS analysis in the lymph node of NC/Nga mice 28
9. FACS analysis in the spleen 33
10. RT-PCR isolated from the facial skin tissues 39
1) Effects of TG and ET on IL-4 and IFN-γ mRNA expression 39
2) Effects of TG and ET on IL-5 and CCR3 mRNA expression 39
11. Effects of TG and ET on IL-6 and IL-13 mRNA expression 41
12. Determination of ratios of CD69+/CD3εand CD19+/CD44by FACS analysis 43
13. Histopathological analysis of lesioned ear skin tissues 44
14. Histopathological analysis of lesioned skin tissues 46
15. Lymphokine assay in the splenocytes 48
Ⅳ. Discussion 50
Ⅴ. Conclusion 59
References 61
Korean abstract 68
Table 1. Prescription of Tang-gwi-eum-za-gagambang (TG) 11
Table 2. Prescription of external spray treatment (ET) 11
Table 3. Cell Content in Lymph Node of NC/Nga Atopy
Dermatitis Model Mice 29
Table 4. Cell Content in Spleen of NC/Nga Atopy Dermatitis Model
Mice 34
Fig. 1. Clinical skin features and severity of dermatitis in NC/Nga mice 20
Fig. 2. Serum IgE and IL-6 elevation and development of
dermatitis in NC/Nga atopy dermatitis model mice 22
Fig. 3. Serum IL-4 and IL-5 elevation and development of
dermatitis in NC/Nga atopy dermatitis model mice 23
Fig. 4. Serum IL-13 and IFN-g elevation and development of
dermatitis in NC/Nga atopy dermatitis model mice 24
Fig. 5. Spleen weight in NC/Nga atopy dermatitis model mice 25
Fig. 6. IgM in NC/Nga atopy dermatitis model mice 26
Fig. 7. Immunoglobulin G1 and G2a in NC/Nga atopy dermatitis model
mice 27
Fig. 8. Effect of TG and ET cotreatment on CD3ε+ and CD19+ (%)
population in NC/Nga atopy dermatitis mouse lymph node
cells 30
Fig. 9. Effect of TG and ET cotreatment on CD4+ and CD8+ (%)
population in NC/Nga atopy dermatitis mice lymph node
cells 31
Fig. 10. Effect of TG and ET cotreatment on CD3ε+/ CD69+and
CD11a+/CD19+ (%) population in NC/ Nga atopy dermatitis
mice lymph node cells 32
Fig. 11. Effect of TG and ET cotreatment on CD3ε+ and CD19+ (%) population in NC/Nga atopy dermatitis mice spleen cells 36
Fig. 12. Effect of TG and ET cotreatment on CD4+ and CD8+ (%)
population in NC/Nga atopy dermatitis mice spleen cells 37
Fig. 13. Effect of TG and ET cotreatment on CD3ε+/CD69+ and
CD11a+/CD19+ (%) population in NC/Nga atopy dermatitis mice
spleen cells 38
Fig. 14. RT-PCR analysis for IL-4 and IFN-γ mRNA 40
Fig. 15. RT-PCR for IL-5 and CCR3 mRNA 40
Fig. 16. Culture supernatant IL-6 and IL-13 production in facial
skin biopsy of atopic dermatitis-like skin lesions in
NC/Nga mice 42
Fig. 17. FACS analysis of cultured facial & neck skin cells in
dermatitis-like skin lesions in NC/Nga mice 43
Fig. 18. Histologic features of ear lesion in NC/Nga mouse 45
Fig. 19. Histologic features of ear lesion in NC/Nga mouse 45
Fig. 20. Histologic features of lesioned skin tissue in NC/Nga mouse 47
Fig. 21. Histologic features of lesioned skin in NC/Nga mouse 47
Fig. 22. IL-4, IFN-γ synthesis in splenocytes from treated with TG
and ET in C57BL/6 mice 49
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