Molecular biological study on ischemic cerebral injury.

缺血性脑损伤的分子生物学研究。

• 批准号: 10470287
• 负责人: KOBAYASHI Hidenori
• 金额: $ 5.25万
• 依托单位: OITA MEDICAL UNIVERSITY (2000)福井医科大学 (1998-1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470287/
• 关键词: Cerebral ichemia; Hippocampus; Monglian gerbil; mRNA; cDNA expression array; cDNA Expression Array; 虚血耐性; differential display; protein kinase II; 14-3-3protein; neuronal pentraxin

The mechanisms of ischemic cell damage are still a matter of debate. Using Atlas cDNA expression arrays, we examined changes of mRNA expression profile in the hippocampus following 5-minute-forebrain ischemia in Mongolian gerbils. Under pentobarbital anesthesia, gerbils were sacrificed by decapitation at 6 hours (n=5) and 2 days (n=5) after ischemia, and sham-operated gerbils (n=5) were sacrificed at 6 hours after surgery. Poly A^+ RNA was isolated from the hippocampal samples in each group. The first step was to reverse transcribe 1 μg of each RNA population using the reagents and [α-^<32>P]dATP.The radioactively labeled, complex cDNA probes were separately hybridized overnight to the Atlas arrays. The Atlas arrays were exposed to the imaging plate of the FUJI bioimaging analyzer. Autoradiography in the control group showed 72 hybridization signals out of 588 cDNA dots. The changes in mRNA expression were classified into 3 patterns ; decrease or disappearance (29 mRNAs), decrease and recovery (11 mRNAs), and increase (32 mRNAs) or new appearance (38 mRNAs). New appearance mRNAs were related to cell-cell communication, protein turn over, stress response protein, and growth factors. It is important to investigate altered mRNA expression profiles following transient forebrain ischemia.

缺血性细胞损伤的机制仍然是一个有争议的问题。利用Atlas cDNA表达阵列技术，研究了蒙古沙土鼠前脑缺血5分钟后海马mRNA表达谱的变化。在戊巴比妥钠麻醉下，沙土鼠在缺血后6 h（n=5）和2d（n=5）断头处死，假手术沙土鼠在手术后6 h处死。从每组的海马样本中分离出Poly A^+ RNA。第一步是使用试剂和[α-^ P]dATP逆转录1 μg每种RNA群体<32>。将Atlas阵列暴露于FUJI生物成像分析仪的成像板。对照组的放射自显影显示588个cDNA点中有72个杂交信号。mRNA表达的变化分为3种模式：减少或消失（29个mRNA），减少和恢复（11个mRNA），增加（32个mRNA）或新出现（38个mRNA）。新出现的mRNA与细胞间通讯、蛋白质周转、应激反应蛋白和生长因子有关。研究短暂性前脑缺血后mRNA表达谱的改变是重要的。

项目成果

Kobayashi H,Matsukawa S,Kobayashi S,Kubota T: "Changes in expression of mRNAs in the gerbil hippocampus following transient forebrain ischemia."Neurol Res. 22・8. 825-831 (2000)

小林 H、松川 S、小林 S、久保田 T：“短暂前脑缺血后沙鼠海马中 mRNA 的表达变化”Neurol Res 825-831。

yashi H, Kitai R, Ido K, Kabuto M, Handa Y, Kubota T, Yonekura Y: "Hemodynamic and metabolic changes following cerebral revascularization in patients with cerebral occlusive diseases."Neurol Res. 21(2). 153-160 (1999)

yashi H、Kitai R、Ido K、Kabuto M、Handa Y、Kubota T、Yonekura Y：“脑闭塞性疾病患者脑血运重建后的血流动力学和代谢变化。”Neurol Res。

Kobayashi H, Matsukawa S, Kodera T, Kabuto M, Handa Y, Kubota T: "Induction of a novel ischemic response gene product in gerbil brain."J Cereb Blood Flow Metab. (Suppl.1.). S303 (1999)

Kobayashi H、Matsukawa S、Kodera T、Kabuto M、Handa Y、Kubota T：“在沙鼠脑中诱导新型缺血反应基因产物。”J Cereb Blood Flow Metab。

Uno H, Kobayashi H, Handa Y, Kabuto M, Kubota T: "Alterations of calcium/calmodulin dependent protein kinase II activity in ischaemia-induced neuronal death and neuronal protection against ischaemia in the gerbil hippocampus."Acta Neurochir (Wien). 141(3)

Uno H、Kobayashi H、Handa Y、Kabuto M、Kubota T：“钙/钙调蛋白依赖性蛋白激酶 II 活性在沙鼠海马缺血诱导的神经元死亡和针对缺血的神经元保护中的变化。” Acta Neurochir (Wien)。

Kobayashi H, Matsukawa S, et al.: "Induction of a novel ischemic response gene product in gerbil brain"J Cereb Blood Flow Metab. Suppl. 1. S303 (1999)

小林 H、松川 S 等人：“在沙鼠脑中诱导新型缺血反应基因产物”J Cereb Blood Flow Metab。