目的:观察不同浓度红景天苷(salidroside)对实验性自身免疫脑脊髓炎(experimental autoimmuneencephalomyelitis,EAE)小鼠模型的作用,并探讨初步的作用机制.方法:建立实验性自身免疫性脊髓炎小鼠模型,免疫当天起给予高中低剂量的红景天苷灌胃,1次/d,共给药35 d.给药完成后,进行神经功能评分,TUNEL染色检测脊髓髓鞘组织的细胞凋亡情况,ATP水平检测试剂盒检测脊髓组织ATP含量水平,免疫印迹法检测线粒体内细胞色素c氧化酶(mito cyto-c)、胞浆中细胞色素c氧化酶(cyto cyto-c)、活化的含半胱氨酸的天冬氨酸蛋白水解酶-3(cleaved cas-3)与活化的含半胱氨酸的天冬氨酸蛋白水解酶-9(cleaved cas-9)蛋白表达水平.结果:红景天苷降低EAE小鼠的神经功能评分,后延发病时间;随红景天苷浓度升高,脊髓髓鞘组织的细胞凋亡数逐渐降低,脊髓组织ATP水平显著升高;EAE小鼠脊髓组织细胞胞浆中的cyto-c蛋白表达显著升高,同时cleavedcas-3与cleaved cas-9蛋白表达显著升高,红景天苷干预后,胞浆中cyto-c蛋白表达显著下降,同时cleaved cas-3与cleaved cas-9蛋白表达下降.结论:红景天苷对缓解实验性自身免疫性脊髓炎症状具有良好的效果,可能通过抑制内源性线粒体通路而降低脊髓组织细胞的凋亡而发挥作用.%Objective:To investigate the effect of different concentrations of Salidroside on experimental autoimmune encephalomyelitis (EAE) mice and study the possible mechanism.Methods:EAE mice model was established and mice were administrated with different concentrations of Salidroside by gavage once a day for 35 days.After the experimental period,neurological function was scored,cell apoptosis in spinal cord myelin was detected by TUNEL staining and ATP level in spinal cord tissue was measured by ATP detection kit.Moreover,the protein expressions of mito cyto-c,cyto cyto-c,cleaved cas-3 and cleaved cas-9 were determined with Western blotting.Results:Salidroside decreased the scores of neurological function in EAE mice and delayed the onset of EAE.The cell apoptosis in spinal cord myelin of EAE mice were inhibited by Salidroside dose-dependently,whereas the ATP level was increased.Moreover,Salidroside administration significantly inhibited the release of cyto-cfrom mitochondria to cytoplasm and decreased the expression of cleaved cas-3 and cleaved cas-9.Conclusion:Salidroside can effectively alleviate the symptoms of EAE,which may be through the inhibition of intrinsic mitochondria-dependent cell apoptosis in spinal cord myelin.
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