梓醇配伍大黄素对实验性自身性免疫性脑脊髓炎小鼠大脑内神经丝蛋白200和髓鞘碱性蛋白的影响

Effects of catalpol combining with emodin on neurofilament protein 200 and myelin basic protein expression with experimental autoimmune encephalomyelitis mice

目的:观察梓醇配伍大黄素对实验性自身性免疫性脑脊髓炎(EAE)小鼠大脑内神经丝蛋白(NF)200和髓鞘碱性蛋白(MBP)表达的影响,讨论梓醇配伍大黄素减轻EAE小鼠大脑内轴突及髓鞘损伤的机制。方法:将80只6~8周龄C57BL/6j小鼠随机分为4组,正常组、模型组、激素组和配伍组各20只,除正常组外,其余3组小鼠背部脊柱旁皮下四点注射抗原髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)诱导产生EAE,观察各组小鼠神经功能缺损严重程度,于发病高峰期(第18天)及缓解期(第40天)取小鼠大脑组织。透射电子显微镜(TEM)观察小鼠大脑髓鞘、轴突微结构病理改变。免疫组化(IHC)技术检测小鼠大脑NF200和MBP表达水平。结果:与模型组比较,配伍组在第21天,27天至39天神经功能功能损伤评分降低(P<0.05)。TEM显示激素组与配伍组髓鞘环形层状结构松散变形程度均较模型组减轻;第18、40天,模型组NF200及MBP积分光密度值显著低于正常组(P<0.01),激素组及配伍组NF200及MBP积分光密度值显著高于模型组(P<0.01,P<0.05)。第40天,激素组MBP积分光密度值显著低于正常组(P<0.05)。结论:梓醇配伍大黄素能够减轻EAE小鼠神经功能缺损,增加潜伏期,减少轴突和髓鞘的损伤。

Objective:To observe the effects and explore the mechanism of catalpol combining with emodin on the expression of neurofilament protein(NF)200 and myelin basic protein(MBP)in the brain of experimental autoimmune encephalomyelitis(EAE)mice.Methods:Eighty C57BL/6j mice of 6 to 8 weeks old were randomly divided into 4 groups,normal group,model group,PA group and EA group.Except the normal group,the other 3 groups were subcutaneous injected with antigens MOG35-55 to induce EAE,and neurological scores were estimated daily.Pathological changes of myelin and axon were observed by transmission electron microscope(TEM).NF200 and MBP were detected by immunohistichemistry(IHC)in brain of mice.Results:Catalpol and emodin could reduce the rate of appearance and behavior change,reduce the mortality rate,prolong the latency of appearance and behavior change,and down regulate neurological function scores of EAE mice.At day 21,27 to 39,the neurological function score decreased compare with model group(P<0.05).TEM showed that the loose deformation of the myelinated annular lamellar structure in PA group and EA group were alleviated compared with the model group.On the 18th and 40th day,the integral optical density(IOD)of NF200 and MBP in the model group was significantly less than that in the normal group(P<0.01).NF200 and MBP in the PA group and EA group were significantly higher than those in the model group(P<0.01,P<0.05).MBP in the PA group was lower than that in the normal group on the day 40(P<0.05).Conclusion:Catalpol and emodin can reduce the nerve function defect in the meantime increase the incubation period,and reduce the damage of axon and myelin.