Expression of Antimicrobial Peptides: RESULTS

18 Jan
2011

Atopic dermatitis1) TEWL measurement between involved and uninvolved skin

For comparisons of TEWL levels between involved and uninvolved sites in AD, measurements were performed 5 times for each site, repeatedly. The mean TEWL level of healthy donor was 7 g/hm2. The mean TEWL levels increased in both un­involved and involved sites than that of healthy donors, especially higher increased in involved sites

2) Expression of LL-37 and hBD-2 mRNA in involved and uninvolved sites of AD using RT-PCR method

Compared to the expression of AMPS of healthy donor, the expressions of hBD-2 and LL-37 were decreased. LL-37 and hBD-2 mRNA expressions more decreased in involved sites than in uninvolved viagra 50 mg

fig1. tr-pcr

Fig. 1. RT-PCR for hBD-2, LL-37mRNA in specimens from uninvolved (U), involved(I) sites and normal control(C) with atopic dermatitis. The expression of hBD-2, LL-37 mRNA were down-regulated in both uninvolved and involved sites, more decreased in involved sites (*p<0.05)

fig2. expression

Fig. 2. Expression of hBD-2, LL-37 proteins in specimens from uninvolved (U), involved (I) sites and normal control (C) with atopic dermatitis by Western blot. The production of hBD-2 in involved skin was markedly decreased compared to that in uninvolved skin. The upper laned are actins.

3) Expression of LL-37 and hBD-2 protein in involved and uninvolved sites of AD using Western blotting

Compared to the expression of AMPS of healthy donor, the expression of hBD-2 and LL-37 were decreased. The production of hBD-2 and LL-37 in involved sites were markedly decreased compared to that in uninvolved sites(Fig. 2, £><0.05).
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Table 1 Comparison of mean TEWL levels between involved and uninvolved sites in atopic dermatitis, average levels of TEWL in age and anatomical site matched normal control was 7 g/hm2

No.

TEWL (g/hm2)

Involved

Uninvolved

>EWL

1

36

22

14

2

24

8

16

3

55

30

25

4

60

47

13

5

60

20

40

6

34

10

24

7

46

30

16

4) IHC for LL-37 and hBD-2 in involved and uninvolved sites of AD

The intensity of stainings of both AMPs in involved skin were significantly weaker than those in uninvolved skin (Fig. 3). The IHC revealed the presence of more abundant hBD-2 and LL-37 in the uninvolved sites than involved sites.

fig3. immunohistochemical

Fig. 3. Immunohistochemical staining For hBD-2 and LL-37 in frozen sections from patients with atopic dermatitis. The IHC confirmed the presence of more abundant hBD-2 and LL-37 in the uninvolved sites than involved sites.

5) Regression curves

For evaluation of the expression of AMPs in according to TEWL levels changes in AD, the subtraction values of AMPs expressions and TEWL levels, which were measured from involved and noninvolved sites of each patient, were calculated, respectively. When these results compared to each other using regression curve, subtraction values of AMPs expressions were more decreased according to increase of TEWL levels. It showed inverse pro­portional relationship (Fig. 4, 5).
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fig4. rna

Fig. 4. RNA expression amounts of hBD-2 (circle) LL-37 (square) and A EWL (triangle)

fig5. regression

Fig. 5. Regression curves A RNA expression amounts of hBD-2 (diamond, solid line), LL-37 (triangle, dotted line) and A EWL (A RNA : mRNA amount of uninvolved site mRNA amount of involved site).

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