NITRIC OXIDE IN THE SKIN AND UVB INFLUENCE

皮肤中的一氧化氮和 UVB 的影响

• 批准号: 07670944
• 负责人: OKAMOTO Hiroyuki
• 金额: $ 1.41万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670944/
• 关键词: NITORIC OXIDE; KERATINOCYTE; SKIN; UV; INFLAMMATORY DISEASE; INOS

Nitric oxide (NO) is an intercellular messenger molecule involved in a variety of biological functions and is generated in mammalian organs by NO synthase (NOS). Inducidle NOS (iNOS) is induced in various types of tissues and cells by endotoxins, interferon-gamma (IFN-gamma), interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha), alone or in combination. In this study, we examined whether iNOS is induced in the epidermis of some skin diseases and in cytokine-stimulated keratinocytes. Isolated rat epidermal cells produced nitrite in the supernatant fluid when stimulated with interleukin-1beta and lipopolysaccharide. Depletion of Ia-positive cells from the epidermal cells slightly eliminated the activity of nitrite production. Homogenates of the stimulated cells exhibited a main band at 150kDa by the Western blot method. The same protein bands were also visible in the human keratinocyte cell line, HSC-1, cultured with the same stimulants. When stained with iNOS-specific monocl … More onal antibodies by the avidin-biotin-peroxidase immuno-histochemical method, the epidermis of some skin diseases, including lichenoid tissue reactions, psoriasis, collagen diseases and atopic dermatitis, showed a positive reaction to the antibodies. Topical medication of steroids decreased the expression with improvement of the disease activity in psoriasis and atopic dermatitis. There was no diffrence in iNOS expression in the epidermis between allergic type and croton-induced irritant type contact dermatitis. Peripheral blood cells had an ablity to stimulate keratinocytes to produce NO in atopic dermatitis patients compared to normal volunteers. These findings suggest that iNOS probably induced in keratincytes by cytokines is relevant to the pathology of some inflammatory skin diseases.The nitrite production by UVB-irradiated kerationocytes and the effects of the NOS inhibitor on ear swelling and sunburn cell formation after UVB irradiation were examined. Nitrite was identified in the supernatant fluid of interleukin 1 (IL-1) -stimulated transformed keratinocyte cell lines. Five mJ/cm^2 UVB augumented the production of nitrite. Nitrite was not generated in the keratinocytes irradiated but not stimulated with IL-1. When BALB/c mice were irradiated with 200mJ/cm^2 UVB,ear swelling and sunburn cells were observed 24 hours after irradiation. When the mice were injected with a NOS inhibitor, N^G-nitro-L-arginine methyl ester (L-NAME), before irradiation, ear swelling was significantly suppressed.Furthermore, such injection also suppressed the formation of sunburn cells. These findings suggest that NO produced in the skin by UVB irradiation contributes to keratinocyte injury as well as acute dermal reactions such as UVB edema. Less

一氧化氮（NO）是一种参与多种生物功能的细胞间信使分子，在哺乳动物器官中由NO合成酶（NOS）产生。诱导型NOS （iNOS）是由内毒素、干扰素- γ (ifn - γ)、白细胞介素-1 （IL-1）和肿瘤坏死因子- α (tnf - α)单独或联合在各种类型的组织和细胞中诱导产生的。在这项研究中，我们检测了iNOS是否在一些皮肤病的表皮和细胞因子刺激的角质形成细胞中被诱导。分离的大鼠表皮细胞在受到白细胞介素-1 β和脂多糖刺激后，在上清液中产生亚硝酸盐。表皮细胞中ia阳性细胞的耗竭轻微地消除了亚硝酸盐产生的活性。Western blot结果显示，受刺激细胞匀浆在150kDa处出现一条主带。同样的蛋白带在用同样的刺激物培养的人角质形成细胞系HSC-1中也可见。用亲和素-生物素-过氧化物酶免疫组化方法对inos特异性单抗进行染色，发现类地衣组织反应、牛皮癣、胶原蛋白疾病和特应性皮炎等皮肤病的表皮对inos特异性单抗呈阳性反应。在银屑病和特应性皮炎患者中，局部类固醇药物治疗可降低表达，改善疾病活动性。过敏型和刺激性型接触性皮炎表皮iNOS的表达无差异。与正常志愿者相比，特应性皮炎患者外周血细胞有能力刺激角质形成细胞产生NO。这些发现提示细胞因子在角质形成细胞中诱导的iNOS可能与一些炎症性皮肤病的病理有关。研究了UVB照射后角质形成细胞产生亚硝酸盐的情况，以及NOS抑制剂对UVB照射后耳部肿胀和晒伤细胞形成的影响。在白细胞介素1 （IL-1）刺激的转化角质细胞细胞系的上清液中鉴定出亚硝酸盐。5 mJ/cm^2的UVB增强了亚硝酸盐的生成。照射后的角质形成细胞不产生亚硝酸盐，IL-1也不刺激。以200mJ/cm^2 UVB照射BALB/c小鼠，24小时后观察到耳肿胀和晒伤细胞。辐照前给小鼠注射NOS抑制剂N^ g -硝基- l -精氨酸甲酯（N^ g -硝基- L-NAME），可明显抑制小鼠耳肿胀。此外，这种注射也抑制了晒伤细胞的形成。这些发现表明，UVB照射在皮肤中产生的NO有助于角质细胞损伤以及急性皮肤反应，如UVB水肿。少

项目成果

K.Mizuno, H.Okamoto, K.Yoneda, K-I.Toda, R.Hattori, Y.Yui, S.Imamura: "Inducible nitric oxide synthase in epidermal keratinocytes." J.Invest.Dermatol.104. 653 (1995)

K.Mizuno、H.Okamoto、K.Yoneda、K-I.Toda、R.Hattori、Y.Yui、S.Imamura：“表皮角质形成细胞中的诱导型一氧化氮合酶。”

K.Mizuno et al: "Inducble nitvic oside Syuthase in epidermal leratinoytes" J. Invest. Derwatol. 104. 653 (1995)

K.Mizuno 等人：“表皮 leratinoytes 中诱导硝基糖苷 Syuthase”J. Invest。

G.Zaipei, K.Yoneda, H.Okamoto and S.Imamura: "Apoptosis with DNA fragmentation in Bowen's disease and sqiamous cell carcinoma." Acta.Histochem.Cytochem.29. 261-263 (1996)

G.Zaipei、K.Yoneda、H.Okamoto 和 S.Imamura：“Bowen 病和鳞状细胞癌中 DNA 碎片导致的细胞凋亡。”

G.Zaipel et al.: "Arghoris with DNA fragmentation in Boweuj diseare and sguawous cell carinowa" Acfa. Histochew. Cytochew.29. 261-263 (1996)

G.Zaipel 等人：“Boweuj diseare 和鳞状细胞 carinowa 中具有 DNA 片段化的 Arghoris”Acfa。

H.Okamoto et al: "Nitric oxide in ultraviolet B radiation-induced keratinocyte in jury in nice" J. Invest. Derwatol. 104. 682 (1995)

H.Okamoto 等人：“尼斯评审团中紫外线 B 辐射诱导的角质形成细胞中的一氧化氮”J. Invest。