腸管上皮細胞における透過性制御機序の解明と臨床への応用

肠上皮细胞通透性控制机制的阐明及临床应用

• 批准号: 19K18354
• 负责人: 尾迫 貴章
• 金额: $ 2.75万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2019
• 资助国家: 日本
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-19K18354/
• 关键词: 虚血再灌流傷害; 細胞接着結合; 細胞密着結合; 腸管透過性; 透過性; タイトジャンクション; 水素ガス; 腸管上皮細胞; 虚血再還流傷害; 医療ガス; 酸化ストレス

過大侵襲下の腸管虚血再灌流傷害は、多臓器不全にまで進展する致死率の高い急性病態である。腸管虚血再灌流傷害では、活性酸素種(ROS)が 腸管上皮透過性亢進作用を主とした腸管虚血再灌流傷害・腸管バリア機能破綻に直接的な役割を演じるとされる。我々は、水素ガスがROSの制 御に重要な役割を果たすことを発信してきた。本研究の目的は、侵襲度を確実に調整出来る腸管虚血再灌流傷害モデルで、腸管上皮細胞の活性 酸素種発現と細胞透過性亢進の関連を検討することで酸化ストレスシグナルの役割を解明し、さらに腸管虚血再灌流傷害を水素ガスにより抑制できるかどうか検討することである。Caco-2細胞をtranswellのapical底に播種 しchamber内 に100μLの培養液を充填し培養。同時にbasal chamber内に600μLの培養液を充填。播種細胞数は1×10*4/cm2~5× 10*5/cm2間で5段階に設定しモデル作成を検討した。虚血再灌流モデルとしての虚血状態はチャンバー内低酸素(1%)と無グルコース培地置換により、再灌流は通常酸素とグルコース含有培地に戻すことで再還流状態とし、虚血再灌流モデルとした。虚血時間は6時間に設定し、介入群は7ppmに調整した飽和水素培地をapical chamber内に虚血開始時点で充填した。腸管透過性は、TEER測定を用いた。低酸素環境暴露終了後から2時間毎に、TEERを測定。暴露開始直前のTEER値との比で検討した。結果、1×10*4/cm2の播種濃度において、介入群はコントロール群に比し低酸素環境暴露終了時点、2時間後、4時間後、6時間後時点のTEER値が有意差を以て高値であった。つまり、虚血開始時点であっても水素ガス投与により腸管虚血再灌流傷害は軽減されており、水素ガス投与による何らかのシグナルによる作用が関与していると推測された。

Under the condition of excessive invasion, the injury caused by blood deficiency and reperfusion, and the progression of multiple organ insufficiency resulted in a high mortality rate and a high incidence of acute disease. Tube deficiency blood reperfusion injury, active acid species (ROS) tube epithelial permeability hyperactivity of the main cause of deficiency blood reperfusion injury tube injury mechanism can break the direct use of labor to perform surgery. We should pay more attention to the ROS system and make sure that the service is very important. In this study, the aim of this study was to make sure that the whole blood was perfused, and that the vascular epithelial cells showed hyperpermeability induced by hyperactivity of active acids. in this study, the aim of this study was to make sure that the whole blood was perfused, and the active acids in the epithelial cells of the tubes showed hyperosmosis. Caco-2 cell transwell apical bottom sow chamber with 100 μ L liquid to fill the culture. At the same time, the basal chamber was filled with 600 μ L incubator. The number of planting cells is equal to 1 × 10*4/cm2~5 × 10*5/cm2. In the fifth section, the temperature is set to make it into a real plant. The deficiency blood was reperfused with low acid content (1%), and the reperfused blood was usually treated with low acid concentration (1%). The reperfused blood usually contains the blood supply, the blood flow state and the blood flow state. The virtual blood time 6 time setting, the intervention group 7ppm training session, and the hydrin culture apical chamber internal deficiency blood start time filling time. The penetration test and TEER test were used to determine the performance. After exposure to the low-acid environment, the temperature and TEER were measured at 2 hours after exposure. Prior to the start of exposure, the TEER exposure was significantly higher than that of the exposure. Results: the results showed that 1 × 10*4/cm2 was more serious than the low acid environment at the time point, 2: 00, 4: 00 and 6: 00 after the exposure of the low-acid environment. The TEER concentration was significantly higher than that of the low-acid environment. At the beginning of hypotension and hypotension, the infusion of water and blood from the tube does not cause damage to the blood of the patients.