逍遥散对肝郁证失眠大鼠行为学的影响及作用机制研究

Study on effect of Xiaoyao powder on behavior of insomnia rats with liver depression syndrome and its mechanism

目的研究逍遥散对肝郁证失眠大鼠的行为学及PI3K/Akt-mTOR信号通路的影响。方法从42只健康的SD大鼠中随机选择6只作为空白组,剩余大鼠采用慢性夹尾刺激和腹腔注射氯苯丙氨酸复合因子造模法进行肝郁证失眠大鼠模型制备。将存活的模型大鼠随机分为模型组、逍遥散高剂量组、逍遥散中剂量组、逍遥散低剂量组、天王补心丹组、地西泮组(西药组),从造模第7天开始,连续灌胃治疗。之后对大鼠宏观体征进行观察,并运用旷场实验对大鼠进行行为学测试;运用Western blot、Real-Time PCR法对大鼠海马中的PI3K、Akt和mTOR蛋白表达进行测定。结果旷场实验结果显示:模型组及天王补心丹组与其余各组相比,大鼠移动距离、跨越的方格数以及活动次数均明显下降(P<0.05);逍遥散高剂量组、西药组与模型组及天王补心丹组相比,移动距离、跨越的方格数以及活动次数增多(P<0.05),PI3K、Akt、mTOR蛋白表达显著上升(P<0.05)。结论逍遥散可以有效提高肝郁证失眠模型大鼠的活跃程度,并可能是通过调控PI3K/Akt-mTOR信号通路达到治疗目的。而中药复方的剂量变化以及药物是否对证会直接影响到治疗效果。

Objective To study the effect of Xiaoyao powder on the behavior and PI3K/AKT-mTOR signal pathway of insomnia rats with liver depression syndrome.Methods Six of 42 healthy SD rats were randomly selected as the blank group,and the rest of the rats were used to established the insomnia model with liver depression syndrome by chronic tail clamping stimulation and intraperitoneal injection of chlorophenylalanine compound factor.The surviving model rats were randomly divided into model group,Xiaoyao powder high,medium and lose dose group,Tianwang Buxin mini-pills group and diazepam group.From the seventh day after the model was established,the rats were treated by continuous gavage.After that,the macroscopic signs of the rats were observed,and the behavioral test and western blot,real-time fluorescence quantitative-PCR were used to detect the PI3K,AKT and mTOR protein expression in the hippocampus of the rats.Results The results of open field experiment showed that:compared with the other groups,the moving distance,the number of squares crossed and the number of activities in the model group and Tianwang Buxin mini-pills group decreased significantly(P<0.05);compared with the model group and Tianwang Buxin mini-pills group,the moving distance,the number of squares crossed and the number of activities in the high dose Xiaoyao powder group and western medicine group increased significantly(P<0.05),and the PI3K/Akt/mTOR protein increased significantly(P<0.05).Conclusion Xiaoyao powder can effectively improve the activity of insomnia model rats with liver depression syndrome,and it may achieve the treatment purpose through regulating PI3K/Akt-mTOR signal pathway.However,the dosage change of traditional Chinese medicine compound prescription and whether the medicine has effect on syndrome will directly affect the therapeutic effect.