摘要
目的 通过嗜银三步染色、甲苯胺蓝染色和 HE染色观察局灶脑缺血 /再灌注早期的神经元缺血改变 ,找出缺血早期可逆与不可逆损伤范围的界限。方法 Wistar大鼠 42只经线栓法制作局灶脑缺血 /再灌注模型。动物分为空白对照组 ,假手术组 ,缺血 2小时不灌注组 (O2 R0 ) ,缺血 2小时再灌注 0 .5小时组 (O2 R0 .5 )、再灌注 2小时 (O2 R2 )、再灌注 4小时组 (O2 R4)和再灌注 2 4小时组 (O2 R2 4) 7组 ,每组各 6只动物。动物经特殊组织处理后进行 HE、TB染色和嗜银三步染色 ,用图像分析仪测量黑色神经元所在范围 (缺血中心区 )。结果 本实验动物模型的成功率为 90 %。嗜银三步染色后正常神经元呈黄色或棕黄色 ,与棕黄色的背景难以区分 ;缺血损伤神经元呈黑色 ,黑色神经元的胞体呈塌陷外观 ,神经元的突起呈螺丝样并有不同程度的崩解 ;黑色神经元所在区域的神经纤维网呈灰色或灰黑色 ,与邻近区域的棕黄色背景有明显的差别 ;皮层黑色神经元在各层分布不同 ,并有柱型集中特征 ;缺血 2小时不灌注组动物用嗜银三步染色就能显示黑色神经元。结论 嗜银三步染色能选择性显示局灶脑缺血的早期缺血神经元 ,黑色神经元所在范围代表了早期的缺血中心区 ,黑色神经元可能是不可逆损伤神经元。黑色神经元的辨认可用于区分早?
Objectives To observe the early neuron ischemic damage in focal cerebral ischemia/reperfusion with histostaining methods of argyrophil Ⅲ (AGⅢ),Toludine blue(TB),and H&E,and to make out the‘separating line'between the areas of reversible and irreversible early ischemic damage. Methods Forty two male Wistar rats were randomly divided into the following groups:pseudo surgical,blank control,O2R0(occluded for 2 hours and reperfused for 0 hour),O2R0 5,O2R2,O2R4,O2R24.There were 6 rats in each group.Rats in experimental groups were suffered focal cerebral ischemia/reperfusion through a nylon suture method .After a special processor for tissue manage,the brain were coronal sectioned and stained with H&E,TB,and AGⅢ.The area where dark neurons dwell in (ischemic core) were calculated with image analysis system. Results The success rate of ischemic model for this experiment is 90%.After being stained with argyrophil Ⅲ method,normal neurons appear yellow or pale brown,which is hardly distinguished from the pale brown background.The ischemic neuron stained black,and has collapsed body and “corkscrew like” axon or dentries,which were broken to some extent.The neuropil in the dark neurons dwelled area appears gray or pale black,which is apparently different from the pale brown neighborhood area.The distribution of dark neurons in cortex varies according to different layers,and has a character of columnar form.The dark neurons present as early as 2 hours ischemia without reperfusion with AGⅢ method. Conclusions AGⅢ stain could selectively display early ischemic neurons,the area dwelled by dark neurons represent early ischemic core.Dark neuron is possibly the irreversibly damaged neuron.Identification of dark neurons could be helpful in the discrimination between early ischemic center and penumbra.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2000年第1期25-29,共5页
Acta Academiae Medicinae Sinicae
基金
国家"九五"科技攻关项目! ( 96-90 6-0 2 -2 1)
关键词
脑缺血
黑色神经元
半暗带
再灌注损伤
focal cerebral ischemia/reperfusion
dark neuron
penumbra
argyrophil Ⅲ method
