急性一氧化碳中毒后大鼠胼胝体损伤的实验研究

Experimental research of injury in the corpus callosm of rats after acute carbon monoxide poisoning

目的探讨急性一氧化碳（CO）中毒后迟发性脑病（DE）大鼠胼胝体损伤情况。方法健康雄性Wistar大鼠，分次腹腔注射CO制备迟发性脑病模型。于中毒后第1、2、3、5、7、14、20天和对照组各时间段取胼胝体，常规制备石蜡病理切片，行HE染色观察细胞形态，变色酸染色观察髓鞘变化。于中毒后第7天和对照组相应时间段每组各8只大鼠取胼胝体，透视电镜观察髓鞘变化。于中毒后第7天和对照组相应时间段每组各8只大鼠取胼胝体，蛋白印迹试验检测髓鞘碱性蛋白（MBP）的相对含量。八臂迷宫实验训练并检测大鼠认知功能。结果光镜下观察见HE染色中毒组出现白质疏松、泡沫细胞形成，胶质细胞肿胀、反应性增生等变化，以中毒后第7天为著。变色酸染色可见中毒组因纤维脱髓鞘而出现淡染区、泡沫细胞形成，以中毒后第7天为著。透视电镜下见胼胝体髓鞘板层结构紊乱，出现松解、断裂。蛋白印迹试验显示，中毒后胼胝体MBP下降。绪论CO中毒后随着时间推移，大鼠胼胝体出现不同程度的神经脱髓鞘改变。

Objective To investigate the injury of the corpus callosum of delayed encephalopathy after acute carbon monoxide （CO） poisoning in the rats. Methods Healthy male Wistar rats were given intraperitoneal injections of CO to make the models of delayed encephalopathy. Corpus callosum was taken from the poisoned rats after 1,2, 3, 5, 7, 14, 20 d separately. To prepare paraffin wax pathological sections, and to perform hematoxylin - eosin staining to observe the changes of cells morphology, and chromotropic acid staining to observe myelin changes. The details of myelin changes were observed under the electron microscope. The relative contents of myelin basic protein （MBP） were detected in the 7 - day group and control group by Western - blot. Eight - arm maze training and testing was used to evaluate the rat＇s function of cognitive and memory. Data was analyzed with SPSS20.0. Results We found that the rats in CO poisoning group appeared white matter loose, empty bubble form, oligodendrocytes, swelling, reactive hyperplasia in the HE dyeing, especially in 7 - day group. In chromotropic acid staining, weakly dyed areas and empty bubbles were seen in poisoning groups for fiber demyelination, especially in 7 - day group. Myelin shelf structure disorder, loose solution and fracture were observed in the corpus callosum under the electron microscope. Western - blot results showed that the relative content of MBP in the poisoning group declined compared with control group. Conclusion As time goes on after CO poisoning, the corpus callosum of rats appeared various degrees of neural demyelination