奔豚汤对慢性束缚应激小鼠焦虑的影响

Anxiolytic Effect of Bentuntang in Mice with Chronic Restraint Stress

目的:实验观察奔豚汤对慢性束缚应激诱导小鼠焦虑模型的影响,从中枢神经递质角度阐述奔豚汤抗焦虑的可能机制。方法:72只小鼠随机分为6组,分别为正常组,模型组,奔豚汤低、中、高剂量组(3.51,10.53,31.59 g·kg^(-1))和阳性药地西泮组(4 mg·kg^(-1)),各组均采用灌胃给药30 d。通过慢性束缚应激建立小鼠焦虑模型;自主活动检测、旷场实验、明暗箱实验和高架十字迷宫实验从行为学分析奔豚汤对焦虑小鼠行为学的保护作用;高效液相结合荧光检测器检测海马谷氨酸(Glu)和γ-氨基丁酸(GABA)含量;蛋白免疫印迹法(Western blot)检测海马内γ-氨基丁酸受体(GABAa-Rα5),GABA转运体-1(GAT-1)和GAT-3蛋白表达。结果:与模型组比较,奔豚汤高剂量可以明显提高旷场中央区域的时间(P<0.05),增加在高架十字迷宫开臂停留时间和进入次数(P<0.05),提高海马内GABA水平(P<0.05),降低Glu水平(P<0.05),且能够明显降低GAT-1和GAT-3蛋白表达(P<0.05)。结论:奔豚汤能够改善慢性束缚应激诱导小鼠焦虑行为,可能是通过下调海马区GAT-1和GAT-3蛋白表达,进而调节中枢神经系统氨基酸类神经递质的含量。

Objective： To investigate the anxiolytic effect of Bentuntang （BTT） in mice with chronic restraint stress and explore its possible mechanism from the perspective of the central neurotransmitter. Method： The 72 mice were randomly divided into 6 groups： saline group, model group, BTT low dose （3.51 g·kg-1 ） , BTT middle dose （10.53 g·kg-l）, BTT high dose group （31.59 g·kg-1） and diazepam group （4 mg·kg-1）. All the drugs were administrated by intragastric administration for 30 days, Chronic restraint stress was used to induce mice anxiety models; open field test, locomotion activity test, elevated plus maze tests and light/dark box test were used to analyze the protective effect of BTT in stressed mice from the behavioral prospective; high performance liquid chromatography （HPLC） with fluorescence detection were used for detection of glutamate （Glu） and gamma aminobutyric acid （GABA） contents; the expression levels of GABAa-Rct5, GABA transporter （GAT-1） and GAT-3 in hippocampus were detected by Western blot. Result： The experimental results showed that as compared with the model group, high dose BTT could significantly improve the time in the central region in open field test （P 〈 0.05 ） , increase the entries and time in open arms in elevated plus maze tests （P 〈 0. 05） ; increase the level of GABA in the hippocampus （P 〈 0.05 ） , and decrease glutamate level （ P 〈 0.05 ）. In addition, the protein expression levels of GAT-1 and GAT-3 in hippocampus were decreased. Conclusion： BTT could improve the chronic restraint stress-induced anxiety in mice, possibly by regulating the contents of amino acid neurotransmitters via decreasing GAT-1 and GAT-3 protein expression levels in the central nervous system .