目的:观察补肾益髓(BSYS)方及其拆方补肾(BS)和化痰活血(HTHX)方对实验性自身免疫性脑脊髓炎(EAE)小鼠的神经保护作用及脑和脊髓神经丝蛋白(NF)200和硫酸软骨素蛋白多糖(CSPGs)表达的影响。方法:雌性C57BL/6小鼠随机分为正常对照组、模型组、醋酸泼尼松组、梓醇组、BSYS组、BS组和HTHX组。各造模小鼠以髓鞘少突胶质细胞糖蛋白(MOG)35-55诱导EAE模型。各治疗组小鼠每日予相应药物灌胃,共40d。醋酸泼尼松和梓醇为阳性对照药物。每日观察小鼠神经功能评分。取小鼠脑和脊髓,HE染色观察其病理变化,透射电镜观察轴突和髓鞘变化,免疫组化法测定NF200和CSPGs蛋白表达。结果:发病第25-40天,BSYS方和BS方明显降低小鼠神经功能评分(P〈0.05,P〈0.01);第37-40天,BSYS方优于BS方(P〈0.05)。BSYS方及BS方可明显减轻小鼠脑和脊髓内炎细胞浸润评分(P〈0.05,P〈0.01);降低髓鞘/轴突面积比值(P〈0.05,P〈0.01),明显减轻髓鞘及轴突损伤;升高NF200蛋白表达(P〈0.05,P〈0.01);下调CSPGs表达(P〈0.05,P〈0.01)。HTHX方只升高NF200蛋白表达(P〈0.05)。结论:补肾益髓方及其拆方补肾方具有明显的神经保护作用。并通过上调NF200及下调CSPGs促进轴突修复及再生,化痰活血方的作用不明显。
Objective: To observe effects of Bushen Yisui Formula(BSYS) and modified Bushen Yisui Formula including Bushen Formula(BS) and Huatan Huoxue Formula(HTHX) on neuroprotection and expression of neurofilament protein 200(NF200) and chondroitin sulfate proteoglycans(CSPGs) in the brain and spinal cord of mice with experimental autoimmune encephalomyelitis(EAE). Methods: Female C57BL/6 mice were randomly divided into normal control group, model group, prednisone acetate(PA) group, catalpol(CA) group, BSYS group, BS group and HTHX group. The mice model with EAE was established by injecting with myelin oligodendrocyte glycoprotein(MOG) 35-55. Mice in treatment groups were received daily gavage administration with the corresponding medicines for 40 days. PA and CA were taken as positive control medicines. The neurological function scores of mice were recorded once a day. The pathological changes and the injuries of axon and myelin in the brains and spinal cords of mice were observed by hematoxylin eosin(HE) staining and transmission electron microscopy(TEM) respectively, while the protein expressions of NF200 and CSPGs were measured by immunohistochemistry(IHC). Results: The neurological function scores of mice were lower in BSYS and BS groups than in model group from days 25 to 40(P〈0.05, P〈0.01). From days 37 to 40, BSYS group was better than BS group(P〈0.05). BSYS and BS significantly reduced the scores of inflammatory cell infiltration in brain and spinal cord of mice(P〈0.05, P〈0.01) and decreased the ratio of myelin/axon area(P〈0.05, P〈0.01), relieved myelin and axonal injury, increased the expression of NF200 obviously(P〈0.05, P〈0.01), meanwhile, significantly reduced CSPGs(P〈0.05, P〈0.01). But HTHX only increased NF200(P〈0.05). Conclusion: BSYS and BS have good effects on neuroprotection in the mice with EAE, and could also promote axonal repair and regeneration by increasing NF200 and reducing CSPGs, and th