舒郁胶囊对大鼠海马原代培养神经元Ca~(2+)-CaM通路的作用

目的观察海马神经元内Ca2+浓度和CaM蛋白水平表达变化,探索复方舒郁胶囊治疗抑郁情绪的微观作用机制。方法采用孤养结合慢性温和刺激复制抑郁情绪大鼠模型,以调肝方药舒郁胶囊及其君药柴胡提取物进行干预,氟西汀作为阳性对照药,制备各组大鼠血清;运用原代无血清培养技术培养大鼠海马神经元,以制备的各组大鼠含药血清孵育神经元;采用荧光探针Fluo-3/AM染色和细胞免疫荧光技术,分别测定海马神经元胞内[Ca2+]i和CaM蛋白表达水平。结果与正常组相比,模型血清组细胞内[Ca2+]i和CaM蛋白表达水平明显降低(P<0.05);各含药血清组神经元胞内[Ca2+]i和CaM蛋白水平较模型血清组明显升高(P<0.05),与正常血清组差异无显著性。结论舒郁胶囊调控海马神经元胞内Ca2+-CaM体系的变化可能是其治疗抑郁情绪的作用机制之一。

Effect of Shuyu capsule on the Ca2+/CaM signal pathway in primarily cultured rat hippocampal neurons

Aim To investigate calcium concentration and the expression levels of calmodulin in hippocampal neurons,thus providing clues for exploring microscopic mechanisms of Shuyu capsule treating depression emotion.Methods Depression emotion model rats were prepared with the method of combining separate feeding with chronic mild stimulus.Shuyu capsule and Bupleurum root,the monarch drug were used for drug intervention,fluoxetine as a positive control.Serum of rats in each group was collected and then added into the medium of vitro serum-free cultured hippocampal primary neurons.Then calcium concentration was measured by fluorescent Fluo-3/AM staining and the expression levels of calmodulin(CaM) were evaluated by immunofluorescence.Results Compared with neurons treated with normal rat serum,calcium concentration and the expression levels of CaM in neurons treated with model rat serum increased significantly(P<0.05).After incubated by those medicated serum,the calcium concentration and the CaM expression levels of neurons were higher than those in model rat serum treatment group(P<0.05),and showed no difference from normal serum treated cells.Conclusion Shuyu capsule regulates the activity of Ca2+-CaM system in hippocampal neurons,which may be one of the mechanisms of treating depression emotion.