白香丹胶囊对PMDD肝气逆证模型大鼠海马NMDAR1亚基蛋白分布及表达的影响

目的：以PMDD肝气逆证模型大鼠为载体，研究PMDD肝气逆证的病机以及白香丹胶囊对本病的干预机制。方法：改进情志刺激为主多因素分段刺激造模法，复制PMDD肝气逆证大鼠模型；用白香丹胶囊和氟西汀分散片予以干预，最后进行旷场实验评价模型；免疫荧光技术和蛋白印迹技术检测大鼠海马CA1、CA3区NMDAR1受体亚基分布情况及表达水平。结果：模型组大鼠与正常组相比较，其旷场实验的运动总距离明显增加（P<0.001）、中央区域的进入次数及停留在中央区域的时间显著减少（P<0.01），海马CA1、CA3区细胞呈现稀疏杂乱的分布状态，且同氟西汀组一样，其NMDAR1蛋白表达量明显降低；白香丹组大鼠与模型组相比较，其旷场实验的运动总距离明显降低（P<0.05）、中央区域的进入次数和停留在中央区域的时间明显增多（P<0.05），而氟西汀组与之相比，除了运动总距离明显降低（P<0.05），其它指标不存在显著性差异，同氟西汀组一样，白香丹组大鼠海马CA1、CA3区细胞分布、排列无显著异常，但其NMDAR1蛋白表达量显著升高（P<0.0001）；氟西汀组大鼠与白香丹组相比较，其NMDAR1蛋白表达量明显降低（P<0.05）。结论：情志刺激为主多因素分段刺激造模法对大鼠学习记忆能力的损伤机制可能与大鼠海马CA1、CA3区神经细胞的减少和NMDAR1亚基的表达量降低有关,白香丹胶囊通过调整NMDAR1蛋白表达量纠正上述异常改变，从而改善大鼠的学习记忆能力。

Effects of Bai-Xiang-Dan Capsule on Distribution and Expression of NMDAR1 Subunit in Hippocampus of PMDD Rat Model with Liver-qi Invasion

The PMDD model rats with liver-qi invasion were used as carriers to explore the pathogenesis and the mechanism of intervention which Bai-Xiang-Dan (BXD) capsule. PMDD rat model with liver-qi invasion was established with stimulation of emotional distress together with multivariate segmentation method. The BXD capsule and Fluoxetine Hydrochloride Dispersible Tablets were used as interventions. The open-field test was used in the evaluation of animal model. The distribution and expression levels of NMDAR1 subunit in the CA1 and CA3 region of hippocampus were detected with fluorescence microscopy and western blot. The result showed that compared with the normal group, the total distance of the model group in the open-field test was significantly increased (P < 0.001); the times of entering the central zone and the staying time in the central zone were significantly reduced (P<0.01); cells in the CA1 and CA3 region of hippocampus showed sparse arrangement and messy state. The protein expressions of NMDAR1 subunit in the model group and the fluoxetine group were significantly decreased. Compared with the model group, the total distance of BXD capsule group in the open-field test was significantly reduced (P<0.05); the times of entering the central zone and the staying time in the central zone was significantly increased (P<0.05). In the fluoxetine group, there were no obvious differences on parameters except the significantly reduced total distance (P<0.05). The arrangement and distribution of cells in the CA1 and CA3 region of hippocampus in the BXD group and fluoxetine group were with no obvious abnormal. However, the protein level of NMDAR1 subunit in BXD group was significantly increased (P<0.0001). Compared with the BXD group, the protein level of NMDAR1 subunit in the fluoxetine group was significantly decreased (P<0.05). It was concluded that the mechanism of injury on rats both in the learning and memory ability which was stimulated by emotional distress together with multivariate segmentation method may be associated with the reducing of cells in the CA1 and CA3 region of hippocampus and the decreasing of expression activity of NMDAR1 subunit, BXD capsule can effectively correct these abnormal changes back to normal for the improvement of learning and memory abilities in rats.