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目的建立实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠模型,观察其组织病理特征。并进一步研究中药有效成分淫羊藿黄酮(epimedium flavone,EF)对该模型病理特征的干预作用。方法采用豚鼠全脊髓匀浆制备抗原免疫Lewis大鼠建立EAE动物模型,每天监测动物体重和行为学变化;应用HE染色观察脑和脊髓炎性浸润、血管袖套形成等变化;Luxol Fast Blue(LFB)染色观察脊髓脱髓鞘变化;电子显微镜观察脊髓髓鞘超微结构;并考察EF对上述指标的干预作用。结果①随着病程进展,EAE模型组大鼠体重明显下降;从第8天起出现行为学异常,神经功能损伤评分逐渐增高,于第12天达峰值。病理形态学检查显示,EAE模型组大鼠脑组织血管周围和脑实质有大量炎性细胞浸润;脊髓腰膨大脊膜出现炎性水肿,并有炎症细胞浸润;脊髓血管周围有大量炎性细胞密集环绕,呈“袖套样”改变;脊髓髓鞘大面积脱失,且髓鞘板层松散。②EF灌胃给药能显著增加EAE模型大鼠体重,延迟发病进程,缓解发病症状。③EF能减轻EAE模型大鼠脊髓腰膨大和脑实质炎性浸润和髓鞘脱失的病理变化。结论 EF能显著降低EAE大鼠行为学评分,改善EAE大鼠大脑和脊髓的炎性脱髓鞘变化,减轻神经细胞结构损伤,从而改善其临床症状。
Objective To establish a rat model of experimental autoimmune encephalomyelitis (EAE) and observe its histopathological features. The effect of epimedium flavone (EF) on the pathological features of this model was further studied. Methods Lewis rats were immunized with guinea pig spinal cord homogenate to establish animal model of EAE. The body weight and behavioral changes were monitored every day. HE staining was used to observe inflammatory infiltration of brain and spinal cord and vascular cuff formation. Luxol Fast Blue (LFB ) Staining was used to observe the changes of demyelination in spinal cord. The ultrastructure of spinal cord myelin was observed by electron microscope. The effect of EF on the above indexes was also observed. Results ①With the progression of disease, the body weight of rats in EAE model group decreased significantly. From the 8th day, the behavioral abnormalities appeared. The score of neurological injury gradually increased and reached its peak on the 12th day. Pathological examination showed that there were a large number of inflammatory cells infiltrating the perivascular and cerebral parenchyma in the brain tissue of rats in EAE model group; inflammatory edema and infiltration of inflammatory cells occurred in the spinal lumbar dural mesencephalon; a large number of inflammatory cells were dense around the spinal cord Surrounding, was “sleeve-like ” change; spinal cord myelin large area loss, and myelin lamellar loose. ②Ef oral administration can significantly increase the body weight of EAE model rats, delay the onset of disease, relieve the symptoms. ③EF can reduce pathological changes of spinal cord lumbar enlargement and parenchymal inflammatory infiltration and demyelination in EAE model rats. Conclusion EF can significantly reduce the behavioral score of EAE rats, improve the inflammatory demyelination of the brain and spinal cord of EAE rats, and reduce the structural damage of nerve cells, thereby improving their clinical symptoms.