| 研究実績の概要 |
To determine the role of NOX in atopic dermatitis (AD), I use house dust mite (HDM)-induced AD model in NC/Nga mouse, which have been reported as AD-mouse model previously. NC/Nga mouse were treated with or without HDM (Dermatophagoides farinae) 2 times per week for 3 weeks. At the end of the treatment, HDM-treated mouse showed AD-like skin lesion with erythema, excoriation and scaling, an increased of transepidermal water loss and scratching behavior compare to control mouse. In this condition, I found that NOX1 is expressed in mouse keratinocytes, but not NOX2. Moreover, the expression of NOX1 in keratinoctyes was increased in AD skin lesion (HDM group).
Further, I examined the expression of NOX1 in human skin of AD compare to normal condition using gene expression microarray analysis of publicly available datasets. I found an increased of NOX1 expression in AD compare to normal human skin, while NOX2 is similar between those conditions. These findings are confirmed in two different cohorts.
These data suggest that NOX1 is expressed in keratinocytes and its expression is increased in AD condition.
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| 今後の研究の推進方策 |
I would like to investigate further the signaling pathway involve in NOX-induced AD skin condition, especially NOX1. In order to reveal this, I will conduct in vitro experiments using human and mouse keratinocytes cell lines. Keratinocytes will be stimulated with HDM and I will measure the production of pro inflammatory cytokines, epidermal permeability barrier, and Nox expression. Using normal human keratinocytes (NHEK) and human epidermal keratinocytes (HaCaT cells), I will measure the expression of COX2 and PGE2 (pro inflammatory cytokines), filaggrin, loricrin, involucrin, and K10 (the major cornified cell envelope-associated proteins), and all Nox isoforms (Nox1, Nox2, Nox3, and Nox4) following HDM treatment.
Next I will examine the involvement of ROS and Nox in HDM-treated keratinocytes. ROS are significantly found to be increased in lesional and peri-lesional skin in AD and its production has been described by NOX in several different tissues and cells, such as gastrointestinal tract, kidney, heart, lung, and skin. Therefore I will investigate whether NOX, especially NOX1, may involve in ROS production by HDM stimulation in keratinocytes and affecting the AD condition.
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