松郁安神方对失眠大鼠下丘脑生物钟基因表达及学习记忆能力的影响

The Effect of Songyu Anshen Prescription on the Expression of Biological Clock Genes and Learning and Memory Ability in the Hypothalamus of Insomniac Rats

摘要: 目的：观察松郁安神方对失眠大鼠下丘脑生物钟基因表达及学习记忆能力的影响。方法：将40只雄性SD大鼠，采用腹腔注射对氯苯丙氨酸(PCPA)建立失眠大鼠模型，造模成功后随机分为模型组、松郁安神方(SYF)组、Luzindole (Luz)组、SYF + Luz组以及阿戈美拉汀组，另设不造模的对照组，每组8只。灌胃1次/d，连续7d。大鼠空间学习记忆能力采用Morris水迷宫实验测试。下丘脑组织CLOCK和Per1蛋白表达水平采用Western Blotting实验检测。结果：同模型组比较，SYF组和SYF + Luz组大鼠的体质量显著增加(P < 0.01)，SYF组大鼠的逃避潜伏期时间显著缩短(P < 0.01)，跨越平台次数显著增加(P < 0.01)，SYF能够显著上调失眠大鼠下丘脑CLOCK蛋白表达(P < 0.01)，下调Per1蛋白的表达(P < 0.01)。同SYF组相比较，Luz组和SYF + Luz组大鼠的逃避潜伏期时间均显著延长(P < 0.01)，跨越平台次数显著减少(P < 0.01)，Luz组和SYF + Luz组大鼠下丘脑CLOCK蛋白均被显著下调(P < 0.01)，Per1蛋白均被显著上调(P < 0.01)。结论：松郁安神方能够改善失眠大鼠的学习和记忆能力，调节失眠大鼠下丘脑CLOCK和Per1蛋白表达变化，恢复失眠大鼠的正常睡眠节律，该作用可以被褪黑素受体拮抗剂阻断，说明其机制可能与调控下丘脑褪黑素受体通路相关。

Abstract: Objective: To observe the effects of Songyu Anshen Prescription on the expression of biological clock genes and learning and memory abilities in the hypothalamus of rats with insomnia. Method: Forty male SD rats were used to establish the insomnia rat model by intraperitoneal injection of p-chlorophenylalanine (PCPA). After successful modeling, they were randomly divided into the model group, the Songyu Anshen Prescription (SYF) group, the Luzindole (Luz) group, the SYF + Luz group and the agomelatine group. Another control group without modeling was set up, with 8 rats in each group. They were gavaged once a day for 7 consecutive days. The spatial learning and memory ability of rats was tested by the Morris water maze experiment. The expression levels of CLOCK and Per1 proteins in hypothalamic tissues were detected by Western Blotting experiments. Result: Compared with the model group, the body weight of rats in the SYF group and the SYF + Luz group increased significantly (P < 0.01), the escape latency time of rats in the SYF group was significantly shortened (P < 0.01), and the number of platform crossings increased significantly (P < 0.01). SYF could significantly up-regulate the expression of CLOCK protein in the hypothalamus of insomniac rats (P < 0.01) and down-regulate the expression of Per1 protein (P < 0.01). Compared with the SYF group, the escape latency time of rats in both the Luz group and the SYF + Luz group was significantly prolonged (P < 0.01), the number of platform crossings was significantly reduced (P < 0.01), and the hypothalamic CLOCK protein of rats in both the Luz group and the SYF + Luz group was significantly down-regulated (P < 0.01). All Per1 proteins were significantly up-regulated (P < 0.01). Conclusion: Songyu Anshen Formula can improve the learning and memory abilities of insomniated rats, regulate the expression changes of CLOCK and Per1 proteins in the hypothalamus of insomniated rats, and restore the normal sleep rhythm of insomniated rats. This effect can be blocked by melatonin receptor antagonists, indicating that its mechanism may be related to the regulation of the hypothalamic melatonin receptor pathway.