脑还丹对快速老化小鼠SAMP8海马细胞内钙离子浓度及突触后致密区蛋白的影响

Effects of Naohuandan on the hippocampal cells [Ca^(2+)]i and postsynaptic density protein in senescence accelerated mice(SAMP8)

目的:观察复方脑还丹对快速老化小鼠SAMP8海马区[Ca2+]i及突触后致密区PSD-95和Shank-1蛋白的影响。方法:将6月龄SAMP8小鼠(雄性)36只,随机分为模型组、脑还丹组和银可络组,每组各12只,SAMR1小鼠12只为正常对照组。治疗组分别以脑还丹浓缩液及银可络灌胃;正常对照组和模型组以同体积双蒸水灌胃,6周后进行行为学测试,测定各种小鼠海马组织细胞[Ca2+]i,采用免疫组化法和计算机图像分析技术测定海马CA1区PSD-95、Shank-1阳性神经元数目及光密度值。结果:Morris水迷宫实验:与模型组相比,正常对照组、脑还丹组和银可络组逃避潜伏期较模型组明显缩短(P<0.05,P<0.01),药物治疗组在原平台象限停留时间明显增加(P<0.05,P<0.001),且脑还丹组小鼠的表现显著优于银可络组小鼠(P<0.05)。海马区[Ca2+]i:模型组较正常对照组海马细胞[Ca2+]i明显增高(P<0.001);脑还丹治疗组及银可络治疗组[Ca2+]i较模型组明显降低(P<0.001)。免疫组化实验:与模型组相比,正常组、脑还丹组及银可络组海马CA1区PSD-95、Shank-1阳性细胞数较之显著增多(P<0.05,P<0.01)。结论:脑还丹对快速老化小鼠SAMP8学习记忆功能具有改善作用,可能与稳定海马区细胞内[Ca2+]i,延缓细胞衰老,促进海马区PSD-95、Shank-1蛋白的表达,恢复细胞突触的结构和功能有关。

Objective： To observe the effects of Naohuandan on the hippocampal cells [Ca2＋]i and postsynaptic density PSD-95 and Shank-1 proteins in senescence accelerated mice（SAMP8）. Methods： 36 SAMP8 6M-mice（male） were randomly divided into model group, Naohuandan group and Yinkeluo group, with 12 mice in each group. 12 SAMP1 mice were in the normal control group. The treatment groups were respectively gavaged with Naohuandan concentrate and Yinkeluo, the model group and normal control group were respectively gavaged with the same volume double distilled water. The behavioral test was observed after 6 weeks, and determined the [Ca2＋]i in hippocampal tissues, and PSD-95, Shank-1 positive cells number and optical density in the hippocampal CA1 region were detected by using immunohistochemical method and computer image analysis technology. Results： Morris water maze test： The escape latency of model group was obviously longer than other three groups（P〈0.05, P〈0.01）, the staying time in the original platform quadrant was obviously increased in treatment groups（P〈0.05, P〈0.001）, and the mice in Naohuandan group were better than these in Yinkeluo group（P〈0.05）. The hippocampus [Ca2＋]i： Compared with the normal control group and treatment groups, the hippocampus [Ca2＋]i in model group was increased obviously（P〈0.001）, while the hippocampus [Ca2＋]i in the treatment groups were decreased obviously（P〈0.001）, there was no significant difference between Naohuandan group and Yinkeluo group. Immunohistochemistry test： Compared with the model group, thehippocampal CA1 region of PSD-95, Shank-1 positive cells number were significantly increased in the normal control group, Naohuandan group and Yinkeluo group（P〈0.05, P〈0.01）. But there was no significant difference between Naohuandan group and Yinkeluo group. Conclusion： Naohuandan can improve the learning and memory ability of SAMP8 mice. The possible mechanism may be related to making the stability of hippocampus [Ca2＋]i, decreasing cell senescence and promoting hippocampus PSD-95, Shank-1 protein expression, restoring cell synaptic structure and function.