Takemoto O, Yoshimine T, Hayakawa T, Fujita T, Nakajima S, Taneda M, Mogami H
Department of Neurosurgery, Hanwa Memorial Hospital, Osaka, Japan.
No To Shinkei. 1989 Apr;41(4):361-5.
Although cerebral infarction is a destructive process of nerve cells and brain tissue, the nature is not exclusively disintegrating but also includes active cellular reaction which may modify the progression of tissue damage. Most prominent cellular reaction in the area surrounding infarction can be recognized as a trophic or proliferative change of glial cells. In the present study we produced a focal cerebral ischemia in Mongolian gerbils and investigated the dynamic change of astrocytes in the brain adjacent to thalamic infarction. Using immunohistochemical methods, astrocytes were identified with the antibody to astroprotein (GFAP) and the DNA synthesizing (S phase) cells were detected with the antibody to bromodeoxyuridine (BrdU). The posterior communicating artery of a gerbil was occluded by coagulation through the trans-tympanic bulla approach under general anesthesia with ketamine hydrochloride (80 mg/kg, i.p.). Thirty min after intravenous administration of BrdU (200 mg/kg), animals were sacrificed by transcardiac perfusion with 75% ethanol on days 1, 2, 3, 5 and 7 post-infarction. Ethanol-fixed, paraffin-embedded blocks were cut coronally into 6 microns-thick sections at the level of dorsal hippocampus. Double-labeled immunohistochemical technique (avidin biotin peroxidase-complex method) was carried out with each antibody using 3,3'-diaminobenzidine tetrahydrochloride and 4-chloro-1-naphthol as chromogens. The population of GFAP-positive cells and their S-phase fraction (the number of BrdU-positive nuclei divided by the number of GFAP-positive cells expressed in per cent, %) were examined. The data demonstrated that the regional GFAP-positive cells increased continuously between days 1 to 5 (105.9 to 528.8 cells/mm2) postinfarction (44.6 cells/mm2 in normal brain).(ABSTRACT TRUNCATED AT 250 WORDS)
虽然脑梗死是神经细胞和脑组织的一种破坏性过程,但其本质并非仅仅是崩解,还包括可能改变组织损伤进程的活跃细胞反应。梗死灶周围区域最显著的细胞反应可被视为神经胶质细胞的营养性或增殖性变化。在本研究中,我们在蒙古沙鼠中制造了局灶性脑缺血,并研究了丘脑梗死灶附近脑内星形胶质细胞的动态变化。采用免疫组织化学方法,用抗星形胶质蛋白(GFAP)抗体鉴定星形胶质细胞,并用抗溴脱氧尿苷(BrdU)抗体检测DNA合成(S期)细胞。在盐酸氯胺酮(80mg/kg,腹腔注射)全身麻醉下,通过经鼓泡途径凝固沙鼠的后交通动脉。静脉注射BrdU(200mg/kg)30分钟后,在梗死后天1、2、3、5和7通过经心腔灌注75%乙醇处死动物。将乙醇固定、石蜡包埋的组织块在背侧海马水平冠状切成6微米厚的切片。使用3,3'-二氨基联苯胺四盐酸盐和4-氯-1-萘酚作为显色剂,对每种抗体进行双重标记免疫组织化学技术(抗生物素蛋白-生物素过氧化物酶复合物法)。检查GFAP阳性细胞的数量及其S期比例(BrdU阳性细胞核的数量除以GFAP阳性细胞的数量,以百分比表示)。数据表明,梗死后天1至5(105.9至528.8个细胞/mm2)区域GFAP阳性细胞持续增加(正常脑为44.6个细胞/mm2)。(摘要截断于250字)