Imataka K, Handa H, Ishikawa M, Hirai O, Kim S H, Yoshida S, Kinuta Y, Kobayashi S
No To Shinkei. 1987 May;39(5):447-53.
The changes of energy metabolism on vasogenic edema have been largely examined using biochemical quantitative assay. However, the relationship between the sequential changes and blood-brain barrier (BBB) breakdown is not well understood. In the present study, the sequential changes of energy metabolism and potassium in relation to BBB breakdown following the cold-induced brain edema were investigated histochemically. Adult male Wistar rats, weighing 200-250g, were anesthesized with pentobarbital and a burr hole was made in the left parietal region. For evaluating the breakdown of BBB, 2.5% Evans blue (EB) was injected 30 min. before injury, except in the 5 min. model in which it was injected at the time of cold injury. An iron-bar precooled in liquid N2 was placed over the surface for 30 seconds and they were frozen in situ in liquid N2 at 5 min., 2 hrs., 6 hrs., 12 hrs., and 24 hrs., after producing the lesion. The frozen brain was sectioned using a precooled saw in the coronal plane. The brain section was placed in liquid N2 bath and illuminated with 366 nm light (UV) from a 200 watt mercury lamp and Corning filter 5840. NADH fluorescence was recorded photographically through Corning filter 3387 and 5562. Regional ATP and potassium content were investigated histochemically in thin sections with luciferine-luciferase method and Macallum's technique, respectively. At 5 min. after cold injury, leakage of EB was limited within the lesion. Potassium and ATP were decreased in the lesion. NADH fluorescence was increased slightly in the cortex around the lesion.(ABSTRACT TRUNCATED AT 250 WORDS)
利用生化定量分析方法，对血管源性水肿时能量代谢的变化进行了大量研究。然而，能量代谢的连续变化与血脑屏障（BBB）破坏之间的关系尚未完全明确。在本研究中，采用组织化学方法研究了冷诱导脑水肿后能量代谢和钾的连续变化与血脑屏障破坏的关系。选用体重200 - 250g的成年雄性Wistar大鼠，用戊巴比妥麻醉，在左顶叶区域钻一个小孔。为评估血脑屏障的破坏情况，除5分钟模型在冷损伤时注射外，在损伤前30分钟注射2.5%伊文思蓝（EB）。将预冷至液氮中的铁棒置于脑表面30秒，在损伤后5分钟、2小时、6小时、12小时和24小时，将大鼠在液氮中进行原位冷冻。用预冷的锯将冷冻的脑在冠状平面切片。将脑切片置于液氮浴中，用200瓦汞灯发出的366nm光（紫外线）和康宁滤光片5840照射。通过康宁滤光片3387和5562，以摄影方式记录NADH荧光。分别采用荧光素 - 荧光素酶法和麦卡勒姆技术，对薄切片进行组织化学研究，测定局部ATP和钾含量。冷损伤后5分钟，EB渗漏局限于损伤部位。损伤部位钾和ATP含量降低。损伤部位周围皮质的NADH荧光略有增加。（摘要截选至250字）
