目的:观察补肾益髓(Bu Shen Yi Sui,BSYS)方及其拆方补肾(Bu Shen,BS)和化痰活血(Hua Tan Huo Xue,HTHX)方对实验性自身免疫性脑脊髓炎(Experimental Autoimmune Encephalomyelitis,EAE)小鼠脑和脊髓中p-Tau和Beta-tubulin的作用。方法:C57BL/6小鼠雌性随机分为正常对照(NC)、模型(MO)、醋酸泼尼松(PA)、梓醇(CA)、BSYS、BS和HTHX组。造模当天与造模后第7天于小鼠背部两侧皮下注射抗原,即50μg髓鞘少突胶质细胞糖蛋白(MOG)35-55、完全弗氏佐剂(CFA)与结核分支杆菌(TB)混合,并在免疫当天和2 d后腹腔注射500 ng的百日咳毒素(PTX)。每天对小鼠进行灌胃,观察其体重及神经功能评分,并于造模第20 d和第40 d分别取脑和脊髓进行病理检测。采用免疫组化法检测p-Tau和beta-tubulin的表达。结果:与MO组比较,BSYS、BS和HTHX组体重与神经功能评分均出现明显上升,HE染色可观察到炎细胞显著减少,核固缩减轻,同时,BSYS、BS和HTHX方均可明显下调p-Tau表达(P<0.05,P<0.01),上调beta-tubulin蛋白表达(P<0.05,P<0.01)。结论:BSYS方及其拆方BS和HTHX均对EAE小鼠有神经保护作用,表现在减少体重丢失,降低神经功能评分,减轻炎炎性细胞浸润,减轻轴突损伤及促进其修复,尤以BSYS全方更为显著。
Objective： To study the effects of Bushen Yisui Capsule（补肾益髓胶囊, BSYSC） on the oligodendrocyte lineage genes（Olig） 1 and Olig2 in C57BL/6 mice with experimental autoimmune encephalomyelitis（EAE） in order to explore the remyelination effect of BSYSC. Methods： The mice were randomly divided into normal control（NC）, EAE model（EAE-M）, prednisone acetate（PA, 6 mg/kg）, BSYSC high-dose（3.02 g/kg） and BSYSC low-dose（1.51 g/kg） groups. The mice were induced by immunization with myelin oligodendrocyte glycoprotein（MOG） 35-55. The neurological function scores were assessed once daily. The pathological changes in mice brains were observed with hematoxylin-eosin（HE） staining and transmission electron microscope（TEM）. The protein expressions of myelin basic protein（MBP）, Olig1 and Olig2 in brains were measured by immunohistochemistry. The m RNA expressions of Olig1 and Olig 2 was also determined by quantitative real-time polymerase chain reaction. Results： Compared with the EAE-M mice,（1） the neurological function scores were significantly decreased in BSYSC-treated mice on days 22 to 40（P〈0.01）;（2） the inflammatory cells and demyelination in brains were reduced in BSYSC-treated EAE mice;（3） the protein expression of MBP was markedly increased in BSYSC-treated groups on day 18 and 40 respectively（P〈0.05 or P〈0.01）;（4） the protein expression of Olig1 was increased in BSYSC（3.02 g/kg）-treated EAE mice on day 40（P〈0.01）. Protein and m RNA expression of Olig2 was increased in BSYSC-treated EAE mice on day 18 and 40（P〈0.01）. Conclusion： The effects of BSYSC on reducing demyelination and promoting remyelination might be associated with the increase of Olig1 and Olig2.
