多发性硬化动物模型神经功能评分的比较与改良

A comparison study of three neurological assessments in the animal model of multiple sclerosis

目的探讨三种神经功能评分方法在实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitis,EAE）模型中的应用情况,并提出最适评价标准。方法 32只雌性C57BL/6小鼠（18~20 g）随机分配为正常对照组（15只）、造模组（17只）。造模组用MOG_（33-35）多肽与弗式完全佐剂等体积混合成乳剂后多点皮下注射制备成EAE模型。遵循双人双盲原则分别使用Kono’s 5分法、Weaver’s 15分法及改良15分法进行神经功能评分,并结合小鼠脊髓H＆E染色结果,探究三种评分方法的评价准确性、症状敏感性、病情相关性。结果据神经功能评分趋势图,改良15分法后期症状波动更明显。对于非典型性发病EAE,改良15分法和15分法的评分更准确。改良15分法和15分法的发病时间显著早于5分法（P〈0.001）,且可反映早期病理改变。改良15分法与高峰期、缓解期病理改变显著相关（高峰P=0.001,缓解P=0.024）,15分法与缓解期病理改变相关（P〈0.05）。结论在实验性自身免疫性脑脊髓炎动物模型中,改良15分法能有效进行细化评分,更准确地评估各时期神经功能改变。

Objective To evaluate the effectiveness of three neurological assessments in experimental autoim- mune encephalomyelitis. Methods Thirty-two female C57BL/6 mice （18-20g） were randomly divided into normal group （n=15） and immune group （n=17）. Immune group were induced subcutaneously in the flank by emulsion consisted of MOG33-35 and complete Freund＇ s adjuvant. Mice were assessed by two investigators for 35 days after immunization in a blinded manner using Kono＇ s 5-point criterion, Weaver＇s 15-point criterion and improved 15-point criterion. Based on H＆E staining, we analyzed the accuracy, sensibility and dependency of three assessments. Results 35-day clini- cal-score plots revealed that the fluctuation of symptoms was more obvious in improved 15-point criterion compared with the other two criteria. 15-point criterion and improved 15-point criterion are more accurate than 5-point criterion in atypical-onset EAE. 15-point criterion and improved 15-point criterion could detect the neurological deficits much earli- er than 5-point criterion （P〈0.001）. Neurological scores assessed by improved 15-point were well correlated with the pathological findings during the peak and remission stage. Conclusion Improved 15-point criterion is better neurological score system than Kono＇ s 5-point criterion and Weaver＇s 15-point criterion because of its accurate assessment of the neurological deficits in experimental autoimmune encephalomyelitis.