糖尿病網膜症と血液網膜関門のタイト結合

糖尿病视网膜病变和血视网膜屏障的紧密连接

• 批准号: 12877280
• 负责人: 澤田 典均
• 金额: $ 1.28万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Exploratory Research
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12877280/
• 关键词: タイト結合; occludin; 血管内皮細胞; バリア機能; アクチン; 血液網膜関門; 血液脳関門; GDNF; neurturin; 血液透過性; TER

血液脳関門(BBB)と内側血液網膜関門(BRB)は,血管透過性が極めて低く,非常に類似した構造を持ち,調節にも同様な分子機構が働いていると推測される。このような低い血管透過性は,細胞間接着装置のひとつタイト結合のバリア機能によって維持されている.血管内皮細胞のタイト結合は,膜蛋白としてoccludinとclaudin5が知られており,特にoccludinは,他部位の血管に比べBBBやBRBの血管内皮細胞に強く発現していることが知られている.そこで本研究では,occludinの発現が極めて低いラット肺血管内皮細胞(Rat Lung Endothelial cell ; RLE)に,C末端にFLAGを付加したrat occludinを導入し,occludinとタイト結合機能およびアクチン細胞骨格との関係を検討した.Occludin geneを導入し樹立したRLE細胞株では,明らかにoccludin蛋白量が増加し,免疫細胞化学的に細胞接着部位に局在していた.しかしタイト結合機能を示すtransendothelial electric resistance (TER)や,イヌリンやマンニトールの透過性に変化を認めなかった.アクチン細胞骨格を調べるため,EGFP標識wild type RLE (w-RLE)とOccludin導入RLE (o-RLE)をco-cultureし,アクチンの分布を観察した.その結果,w-RLEとw-RLEの接着部位にアクチンの集積を認めないが,o-RLEとo-RLE, o-RLEとw-RLEの接着部位にはアクチンの集積が認められた.逆に,アクチン細胞骨格を破壊すると,occludinは細胞接着部位から消失した.以上から,occludinは,タイト結合のバリア機能の調節より,アクチン細胞骨格とタイト結合を連結する機能を担っていると考えられた.

The blood cell (BBB) has a blood network gate (BRB), the vascular permeability is extremely low, and the type of blood vessel permeability is similar to that of the device. Low vascular permeability is detected, and then a device is installed between the cells to ensure the maintenance of vascular permeability in combination with the mechanism of vascular permeability. When the endothelial cells were combined, the membrane protein of occludin claudin5 was more sensitive than that of BBB BRB, especially occludin. In this study, occludin showed low levels of pulmonary vascular endothelial cells (Rat Lung Endothelial cell). RLE), the C-terminal FLAG plus rat occludin was added into the cell, and the combination of occludin was used to detect the cell cycle. Occludin gene was introduced into the cell line of RLE cell line, the occludin protein content was increased, and the cellular junction site of immunocytochemistry was involved in the cell cycle. The combination machine is capable of displaying transendothelial electric resistance (TER) messages, and the transmission mechanism is sensitive to the transmission of information. The EGFP tag wild type RLE (w-RLE) Occludin enters the RLE (o-RLE) co-culture, and the distribution distributes data. The results showed that the w-RLE w-RLE was followed by the collection of the w-RLE, the o-RLE of the o-RLE, the w-RLE of the cluster, and the location of the cluster. In the opposite direction, the cellular bone lattice is broken, and the occludin cells disappear. The combination of the above information, occludin, computer,

项目成果

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