Development of a new photo-aging model system: A quick evaluation method for photo-damage utilizing elastin-promoter/green fluorescence protein (GFP) reporter gene transfected dermal fibroblasts

开发新的光老化模型系统：利用弹性蛋白启动子/绿色荧光蛋白（GFP）报告基因转染的真皮成纤维细胞的光损伤快速评估方法

• 批准号: 11557062
• 负责人: KON Atsushi
• 金额: $ 3.2万
• 依托单位: Hirosaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557062/
• 关键词: photo-aging; elastin promoter; solr elaslosis; ultraviolet; fibroblast; sunscreen

For the purpose of prevention of photo-aging such as deep wrinkle formation, a great number of sunscreens against broad-band spectrum of solar ultraviolet (UV) has been produced. However, in vivo evaluation of sunscreens for anti-photoaging requires long time. In the present experiments, a new evaluation system have been developed. Cultured mouse fibroblasts (3T3) were transfected with human-elastin promoter linked to green fluorescence protein (GFP) reporter gene. After exposure to UVB, green fluorescence intensity of GFP was observed. In addition, after injection of the cells into the mouse skin, fluorescence intensity was quantified using spectrofluorometer with fiber optics (the SPEX FluoroMax-3 of Jobin Yvon Inc.). The cultured cells expressed green fluorescence in UVB-dose dependent manner. The fluorescence of injected cells was measurable from skin surface. These results indicate that quantitative measurement of fluorescence intensity of injected cells to the mice skin might be useful for early evaluation of solar elastosis formation and protective effect of sunscreens.

为了防止光老化（例如深层皱纹形成），已经生产了大量针对宽带太阳紫外线（UV）光谱的防晒霜。然而，防晒霜抗光老化的体内评价需要很长时间。在目前的实验中，开发了一种新的评估系统。培养的小鼠成纤维细胞 (3T3) 用与绿色荧光蛋白 (GFP) 报告基因连接的人弹性蛋白启动子转染。暴露于UVB后，观察到GFP的绿色荧光强度。另外，将细胞注射到小鼠皮肤后，使用具有光纤的分光荧光计(Jobin Yvon Inc.的SPEX FluoroMax-3)对荧光强度进行定量。培养的细胞以 UVB 剂量依赖性方式表达绿色荧光。注射细胞的荧光可从皮肤表面测量。这些结果表明，定量测量注射到小鼠皮肤的细胞的荧光强度可能有助于早期评估日光弹性组织形成和防晒霜的保护效果。

项目成果

Kon A, Vindevoghel L, Hashimoto I, Uitto J, Mauviel A: "TGF-β/Smad and TNF-a /NF-KB upregulate type VII collagen gene expression additively at the transcriptional level"Molecular Medicine: Novel Findings of Gene Diagnosis, Regulation of Gene Expression, a

Kon A、Vindevoghel L、Hashimoto I、Uitto J、Mauviel A：“TGF-β/Smad 和 TNF-a/NF-KB 在转录水平上调 VII 型胶原蛋白基因表达”《分子医学：基因诊断的新发现》，基因表达的调控

Hashimoto I, Kon A, Tamai K, Uitto J: "Diagnostic dilemma of "sporadic" cases of dystrophic epidermolysis bullosa : a new dominant or mitis recessive mutation?"Exp Deimatol. 8. 140-142 (1999)

Hashimoto I、Kon A、Tamai K、Uitto J：“营养不良性大疱性表皮松解症“散发”病例的诊断困境：一种新的显性突变或 mitis 隐性突变？”Exp Deimatol。

Tamai K, Murai T, Mayama M, Kon A, Nomura K, Sawamura D, Hanada K, Hashimoto I, Shimizu H, Masunaga T, Nishikawa T, Mitsuhashi Y, Ishida-Yamamoto A, Ikeda S, Ogawa H, McGrath JA, Pulkkinen L, Uino J: "Recurrent COL7AI mutations in Japanese patients with d

玉井 K、村井 T、真山 M、Kon A、野村 K、泽村 D、花田 K、桥本 I、清水 H、增永 T、西川 T、三桥 Y、石田山本 A、池田 S、小川 H、麦格拉斯 JA、

Sawamura D, Ina S, Itai K, Meng X, Kon A, Tamai K, Hanada K, Hashimoto I: "In vivo gene introduction into keratinocytes using jet injection"Gene Ther. 6. 1785-1787 (1999)

Sawamura D、Ina S、Itai K、Meng X、Kon A、Tamai K、Hanada K、Hashimoto I：“使用喷射注射将基因导入角质形成细胞体内”Gene Ther。

Sawamura D, Xianmin M, Ina S, Kon A, Tamai K, Ohe Y, Hashimoto I: "Expression vector with DNA of bovine papilloma virus 1 for keratinocyte gene therapy"J Dermatol Sci. 23. 111-116 (2000)

Sawamura D、Xianmin M、Ina S、Kon A、Tamai K、Ohe Y、Hashimoto I：“用于角质形成细胞基因治疗的牛乳头瘤病毒 1 型 DNA 表达载体”J Dermatol Sci。