Expression and function of tight junction proteins in retinal microvascular endothelial cells in the rat model of diabetic retinopathy

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K09415
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56060:Ophthalmology-related
• Research Institution: Sapporo Medical University
• Principal Investigator: Inatomi Shuichiro 札幌医科大学, 医学部, 講師 (10437999)
• Co-Investigator(Kenkyū-buntansha): 大黒 浩 札幌医科大学, 医学部, 教授 (30203748) ; 日景 史人 札幌医科大学, 医学部, 講師 (30837547)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: Diabetic retinopathy / tight junction

Outline of Final Research Achievements

We completed the isolation and culture of cerebral microvascular endothelial cells from the brain of a diabetic model rat, and confirmed that the intercellular adhesion molecule, Occludin, and the tight junction-related molecule, Claudin-5 (Cld-5), are co-localized. It was also confirmed that when cAMP is allowed to act on cerebrovascular endothelial cells, transepithelial electrical resistance, which is an index of tight junction barrier function, increases 3 to 4 times. We also found that glial cell line-derived neurotrophic factor (GDNF) enhances the tight junction barrier function of cerebrovascular endothelial cells for nerve cells secreted from glial cells. We also confirmed that GDNF has the effect of enhancing the barrier function of retinal blood vessels.

Free Research Field

Diabetic retinopathy

Academic Significance and Societal Importance of the Research Achievements

本邦における中途失明原因疾患の第二位である糖尿病性網膜症に関して、本研究では糖尿病モデルラットから網膜血管内皮細胞と同様に強固なタイト結合を有する初代微小脳血管内皮細胞を分離・培養し、その内皮細胞バリア機能を制御するタイト結合関連分子の発現と局在等の形態とバリア機能（透過性または抵抗）を胞間接着分子であるOccludinとタイト結合関連分子であるClaudin-5(Cld-5)を中心に検討した。本手技を用いることにより今後も各種モデル動物からの初代微小脳血管内皮細胞を利用して、さまざまな疾患における血管透過性及び抵抗のメカニズムを明らかにすることが可能と考えられた。