Study of Regulatory Mechanism of Various Growth Factors in Human Epidermal Cells and Fibroblasts

人表皮细胞和成纤维细胞中多种生长因子的调控机制研究

• 批准号: 63480245
• 负责人: YOSHIKAWA Kunihiko
• 金额: $ 4.1万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480245/
• 关键词: Epidermal keratinocyte; Growth and Differentiation; TGF-beta; TGF-alpha; IL-6; Psoriasis vulgaris; PD-ECGF; Involucrin; cーmyc; antiーsense cーmyc; PDーECGF; ケラチノサイト; TGFーβ; TGFーα; EGF; ILー6; オ-トクリン; 乾癬; 表皮細胞; 無血清培養; プラスミノーゲンアクチベーター; プラスミ

(1) Effects of TGF-beta on growth and differentiation of cultured human keratinocytes : TGF-beta inhibited human keratinocyte growth over 0.25 ng/ml and arrested in G_1/G_0 phase of cell cycle. This growth arrest was reversible unless incubation duration is too long.(2) Roles of TGF-alpha in normal and psoriatic epidermis : The localization of TGF-alpha in normal and psoriatic epidermis was examined immunocytochemically. TGF-alpha was observed mainly in the upper layer of normal epidermis. In contrast, TGF-alpha was present intensely through the whole epidermis except basal layer in psoriatic epidermis. This result indicates the increase of TGF-alpha in psoriatic epidermis compared to normal epidermis. To further confirm the increase of TGF-alpha, the amount of TGF-alpha in extracts of normal and psoriatic epidermis was measured by ELISA. The amount TGF-atpha in the extracts of normal epidermis was 1.45 <plus-minus> 1.06 ng/g of wet tissue, while the amount in psoriatic involved epidermis was 6.71 <plus-minus> 0.75ng/g of wet tis(3) Effects of IL-6 on growth and differentiation of cultured human keratinocytes : IL-6 stimulated human keratinocyte growth. Immuno-cytochemical study showed that I1-6 and IL-6 receptor were present in basal layer of epidermis. This suggests that IL-6 possibly stimulates keratinocyte growth in autocrine manner. IL-6 has no clear effect on differentiation of human keratinocytes.(4) Effects of platelet-derived endothelial cell growth factor (PD-ECGF) on growth and differentiation of cultured human keratinocytes : PD-ECGF stimulated human keratinocyte growth. However, PD-ECGF has no clear effect on differentiation of human keratinocytes.(5) A new method for quantifying keratinocyte differentiation using immuno-fluorescent staining of involucrin and cyto-fluorography was established.

(1)TGF-β对体外培养的人角质形成细胞生长和分化的影响：TGF-β在0.25ng/ml以上抑制人角质形成细胞的生长，并使细胞周期阻滞于G_1/G_0期。这种生长停滞是可逆的，除非孵育时间过长。(2)TGF-α在正常和银屑病表皮中的作用：免疫细胞化学检测TGF-α在正常和银屑病表皮中的定位。TGF-α主要在正常表皮的上层观察到。相比之下，在银屑病表皮中，TGF-α强烈地存在于除基底层之外的整个表皮中。该结果表明与正常表皮相比，银屑病表皮中TGF-α的增加。为了进一步证实TGF-α的增加，通过ELISA测量正常和银屑病表皮提取物中TGF-α的量。正常人表皮提取物中TGF-α的含量为1.45 <plus-minus>± 0.6ng/g湿组织，银屑病患者表皮提取物中TGF-α的含量为6.71 ± <plus-minus>0.75ng/g湿组织。（3）IL-6对培养的人角质形成细胞生长和分化的影响：IL-6促进人角质形成细胞的生长。免疫细胞化学研究表明，表皮基底层有I_（1 -6）和IL-6受体。这表明IL-6可能以自分泌方式刺激角质形成细胞生长。IL-6对人角质形成细胞的分化无明显影响。(4)血小板衍生内皮细胞生长因子（PD-ECGF）对培养的人角质形成细胞生长和分化的影响：PD-ECGF刺激人角质形成细胞生长。然而，PD-ECGF对人角质形成细胞的分化没有明显的影响。(5)建立了一种新的表皮免疫荧光染色和细胞荧光照相定量角质形成细胞分化的方法。

项目成果

橋本 公二,吉川 邦彦他(分担): "“皮膚と細胞成長因子" 「現代皮膚科学大系年刊版」" 中山 書店, 14 (1990)

Koji Hashimoto、Kunihiko Yoshikawa 等（撰稿人）：“‘皮肤和细胞生长因子’‘现代皮肤病学年度版’”Nakayama Shoten，14 (1990)

橋本公二: "Immune mechanism in cutaneous disease"In vitro studies of acantholysis of keratinocytes induced by pemphigus antibody"" MARCEL DEKKER,Inc., 277-290 (1989)

桥本幸二：“皮肤疾病的免疫机制”天疱疮抗体诱导的角质细胞棘层松解的体外研究”MARCEL DEKKER,Inc., 277-290 (1989)

K.Hashimoto: "Characterization of Keratinocyte plasminogen activator inhibitors and demonstration of the prevention of pemphigus IgG-induced acantholysis by purified plasminogen activator inhibitor." Dermatology Digest. 9. 14-15 (1989)

K.Hashimoto：“角质形成细胞纤溶酶原激活剂抑制剂的表征以及纯化纤溶酶原激活剂抑制剂预防天疱疮 IgG 诱导的棘层松解症的演示。”

Matsumoto,K.: Exp.Cell Res.

Matsumoto,K.：实验细胞研究。

H. Okumura, K. Matsumoto, K. Hashimoto and Y. Yoshikawa: "A new method for quantifying keratinocyte Differentiation immuno-fluorescent staining of involucrin and cytography." Exp. Cell Res.192. 647-650 (1991)

H. Okumura、K. Matsumoto、K. Hashimoto 和 Y. Yoshikawa：“一种定量角化细胞分化免疫荧光染色外皮蛋白和细胞学的新方法。”