網膜神経細胞の虚血・再潅流傷害時に観察される細胞周期の異常回転と細胞死との関係

视网膜神经元缺血/再灌注损伤过程中细胞周期轮转异常与细胞死亡的关系

• 批准号: 17790174
• 负责人: 坂本 謙司
• 金额: $ 1.66万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17790174/
• 关键词: 薬理学; 脳神経疾患; シグナル伝達

7〜8週齢のSD系雄性ラットをケタミン(90mg/kg)+キシラジン(10mg/kg)で麻酔し,電気毛布で体温を37℃に維持した.片眼の前眼房に,ラットの眼の位置から約175cmの高さの位置に置いた生理食塩水入りのタンクと接続した注射針を刺入し,眼内に130mmHgの圧力を負荷し,網膜虚血を誘発した.網膜虚血を60分間負荷した後,前眼房から注射針を抜くことにより再灌流した.反対側眼は無虚血対照とした.無虚血対照および再灌流24時間後の網膜組織を採取し,ウエスタンブロットによってサイクリンD1,cyclin-dependent kinase(CDK)2,4およびp16やp27などのCDKインヒビタータンパク質をはじめとする細胞周期調節因子の発現量の変化を検討したところ,再灌流24時間後の網膜組織では,無虚血対照と比較し,サイクリンD1およびCDK4の発現が増加しており,また,p16およびp27の発現が減少していることが明らかとなった.この結果から網膜虚血・再灌流傷害にCDKの活性化による細胞周期の異常進行が関与している可能性が考えられたので,次に,CDK阻害薬が網膜虚血・再灌流傷害に与える影響を検討した.同様の方法でラットに網膜虚血・再灌流を負荷し,再灌流7日後に眼球を摘出した.固定後,パラフィン包埋し,薄切組織切片を作成する.再灌流7日後に摘出した眼球の切片はH-E染色を行い,神経細胞の脱落を評価した.虚血15分前にCDK阻害薬であるロスコビチン(3mg/kg)やCDK4 inhibitor(1mg/kg)を投与した群と溶媒を投与した群とを比較したところ,これらのCDK阻害薬は網膜虚血・再灌流により惹起される視神経細胞死を抑制することが明らかとなった.

7-8-year-old SD male anaesthesia (90mg/kg) + maleic acid (10mg/kg) hemp, electric woolen cloth temperature 37 ℃ to maintain temperature. In one eye, the anterior chamber of the eye, the position of the eyes, the position of the eyes, the position of the eyes. After 60 minutes of network deficiency blood loss, the anterior chamber was injected with reperfusions in the anterior eye chamber. In turn, the eyes are full of deficiency and blood is shining on the eyes. After 24 hours of reperfusion, the network tissue was harvested. After 24 hours of reperfusion, the network tissue was harvested. After 24 hours of reperfusion, the network tissue was collected. After 24 hours of reperfusion, the network tissue was collected after 24 hours of reperfusion. After 24 hours of reperfusion, cyclin dependent kinase (CDK) 2, p16, p27, p27, Please tell me that you are not aware of the increase in the number of people in CDK4. Results the results showed that the possibility of activation of CDK cell cycle was evaluated by the possibility that CDK blocked the injury of network virtual blood by reperfusion. the results showed that the damage caused by network virtual blood reperfusion caused by network virtual blood reperfusion. the possibility of cell cycle analysis was evaluated. The same method was used to investigate the reperfussion of epineurial deficiency blood, and the eyeball was removed after 7 days of reperfusion. After fixation, the tissue sections were embedded and thinly cut into thin sections. After 7 days of reperfusion, the eyeball was removed and the cells were exfoliated and stained with Hmure E. Before 15 minutes of blood deficiency, CDK blocked cell death, and so on. 15 minutes before the deficiency blood, the CDK blocked the blood deficiency of the network membrane and reperfused the blood. 15 minutes before the deficiency blood, the CDK blocked the blood deficiency of the network membrane and reperfused it.

项目成果

Inducible nitric oxide synthase inhibitors abolished histological protection by late ischemic preconditioning in rat retina.

• DOI: 10.1016/j.exer.2005.08.011
• 发表时间: 2006-03
• 期刊: Experimental eye research
• 影响因子: 3.4
• 作者: K. Sakamoto;Yuzuru Yonoki;Y. Kubota;M. Kuwagata;M. Saito;T. Nakahara;K. Ishii

Involvement of the betagamma subunits of G proteins in the cAMP response induced by stimulation of the histamine H1 receptor.

G 蛋白的 betaamma 亚基参与刺激组胺 H1 受体诱导的 cAMP 反应。

• 发表时间: 2005
• 期刊: Naunyn-Schmiedeberg's Archives of Pharmacology 372(2)
• 作者: Maruko T;Nakahara T;Sakamoto K;Saito M;Sugimoto N;Takuwa Y;Ishii K.
• 通讯作者: Ishii K.