創傷治癒における各種増殖因子が及ぼす細胞内シグナル伝達解析と臨床応用に関する研究

多种生长因子在伤口愈合中的细胞内信号转导分析及临床应用研究

• 批准号: 16790622
• 负责人: 安部 正敏
• 金额: $ 1.28万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16790622/
• 关键词: コラーゲンゲル; 創傷治癒; b-FGF; G蛋白; Rhoファミリー

前年度、生体内において創傷治癒に深く関与すると考えられる塩基性線維芽細胞増殖因子(b-FGF)ついてのヒト線維芽細胞含有コラーゲンゲルを用いた検討結果より、b-FGFは創傷治癒初期段階モデルであるFloating matrices contraction(FMC)のみを惹起し、晩期段階のモデルであるStressed matrices contraction(SMC)は引き起こさないことを明らかにした。また、b-FGFによるFMCにはRac→Rho→Rhoキナーゼ(ROCK)のシグナル伝達系が関与する可能性が示唆された。そこで、今年度はb-FGFが惹起するFMCの更なるシグナル伝達系を明らかにする目的でPI3Kの阻害剤であるLY294002を加えて検討を行なった。その結果、b-FGFにより惹起されるFMCはLY294002添加によりその効果が阻害された。また、2次元での細胞形態に与えるb-FGFの影響をファロイディン染色により検討したところ、b-FGF添加によりヒト線維芽細胞はラメロポディアの構造を呈しその先端部には接着斑の構成蛋白の一つであるvinculinの発現がみられた。さらに同様の検討において、LY294002およびRhoキナーゼ阻害剤であるY27632の影響を調べたところ、いずれの阻害剤も線維芽細胞の形態変化をもたらし、接着斑の形成も阻害した。b-FGF添加によりヒト線維芽細胞にラメロポディアがみられたことは、Rac活性化により細胞遊走が引き起こされる可能性を示唆する。そこで、Racの活性化を検討するためpull down法を用いてGTP結合型(活性型)Racについて検討したところ、ヒト線維芽細胞はb-FGF刺激後5分でRac活性化が起こり、その活性化はその後30分続いた。更にwound healing assay法にて線維芽細胞遊走能を検討したところ、b-FGFは100ng/mlをピークに線維芽細胞遊走を亢進させた。以上および前年度の研究結果を考え合わせると、b-FGFはPI3K→Rac→Rho→Rhoキナーゼ(ROCK)のシグナル伝達系を介してコラーゲンゲル収縮および細胞遊走を惹起し、皮膚真皮創傷治癒を促すことが明らかとなった。

In the previous year, in vivo and in vivo, there was a study on the basic maintenance of germ cell proliferation factor (b-FGF) in vivo and in vivo. The germ cell contains the results of the treatment of cancer cells, the results of the results of b-FGF treatment, the detection of Floating matrices contraction (FMC) in the early stage of treatment, and the risk of infection. The Stressed matrices contraction (SMC) will be introduced in the following stages. B-FGF, b-FGF, FMC, Rho, Rho, ROCK, ROCK, possibility, possibility, This year's "b-FGF" and "FMC" have caused a lot of trouble. This year, there is a clear understanding that the purpose is to prevent PI3K from harming people, LY294002, and to increase the number of people who do so. The result shows that b-FGF has caused a lot of trouble to FMC the LY294002 to prevent the damage. The cell morphology, the second b-FGF and the second cell morphology were detected by staining, the b-FGF was added to the cell line, the cell morphology was analyzed, and the protein was detected at the end of the cell, and then the protein was detected by vinculin. In the same way, you can use the same information, LY294002, Rho, block, stop, stop, go, go. B-FGF adds a number of positive cell lines, such as the number of cells, and the number of cells activated by Rac. The activation of Rac and the activation of Rac were detected in the pull down method. The activation of Rac was induced at 5 minutes after stimulation with the combination of active Rac and b-FGF, and 30 minutes after the activation. Using wound healing assay method to maintain germ cell line, b-FGF cell line to maintain germ cell line and b-FGF cell to increase the activity of germ cell. In the previous year, the results of the previous year's study showed that the results of the study were combined with the results of the previous year's study, and the results of the previous year's study showed that the results of the previous year's study showed that the results of the previous year's study combined with the results of the previous year's study, the results of the previous year's study showed that the results of the previous year's study showed that the results of the previous year's study combined with the results of the previous year's study, the results of the previous year's study showed that the results of the previous year's study were combined with the results of the previous year's study, and the results of the previous year's study showed that the results of the previous year's study were combined with the results of the previous year's study, and the results of the previous year's study.