| 研究実績の概要 |
First, we evaluated the H2S level in AD patients by measuring the H2S concentration in the serum of healthy (control) and AD patients using ELISA. Serum was collected from 11 AD patients (8 males, 3 females, mean ± SEM age, 38.64 ± 3.22), and 11 controls (5 males, 6 females, mean ± SEM age, 36.55 ± 1.65). We found that AD patients (mean ± SEM, SCORAD 48.5 ± 5.96, VAS 62.75 ± 7.83) had significantly higher concentration of serum H2S compared to control. H2S is now known to be synthesized endogenously in mammals. The production of H2S has been attributed to three enzymes: cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfur-transferase (MPST). The regulation of H2S production occurs at the level of expression and activity of its enzymes. However, the expressions of these enzymes in the skin or keratinocytes (KCs) are unknown so far, therefore we examined their expression in human normal skin and AD lesion. By immunofluorescence modality, we found that the CBS, CSE, and MPST were expressed in human KCs, and all of them were increased in the lesional skin of AD patients compared with that in controls. For all human data, AD was diagnosed clinically based on Hanifin and Rajka criteria.
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| 今後の研究の推進方策 |
We would like to investigate further the effect of gasotransmitter donors in keratinocytes and dorsal root ganglia (DRG) by usin g in vitro experiments. Mouse or human keratinocytes cell line and DRG primary culture will be used to evaluate the production o f gasotransmitter in cultured keratinocytes or DRG, with or without HDM stimulation. Next, with the addition of gasotransmitter donor and HDM, we will evaluate the production of pro inflammatory cytokines or itch-related cytokines , and semaphorin 3A, and NGF, either at mRNA level by real time PCR or protein level by ELISA. Furthermore, it will important also to elucidate the signa ling pathway being involved in these phenomena, e.g. MAP kinase, NF-kB, etc by western blotting analysis
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