朱砂安神丸对条件性恐惧模型大鼠单胺类神经递质表达及其神经元c‐Fos蛋白表达的影响

Effect of ZhushaAnshen pill on expression of monoamine neurotransmitters and c⁃Fos protein in neurons of rats with conditioned fear

目的基于BLA内单胺类神经递质和神经元c‐Fos蛋白表达的变化,探讨朱砂安神丸拮抗大鼠条件性恐惧的作用机制。方法SD大鼠探针植入手术后恢复3 d。随机分为6组:恐惧习得空白组、恐惧习得模型组、恐惧习得给药组(0.81 g/kg)、恐惧消退空白组、恐惧消退模型组、恐惧消退给药组(0.81 g/kg)。于早7点给药组给予朱砂安神丸混悬液(0.09 g/mL,9 mL/kg),各空白组和模型组给予相同体积的双蒸水,每日1次,连续7 d。末次给药后开始进行条件性恐惧动物模型的复制及消退实验,并单笼饲养。消退实验持续7 d,同时给药。通过在体微透析技术和电化学检测器(HPLC‐ECD)联合使用,检测大鼠基底外侧杏仁核(BLA)中单胺类神经递质多巴胺(DA)、去甲肾上腺素(NE)、5‐羟色胺(5‐HT)的含量;以c‐Fos蛋白免疫组化测定大鼠BLA神经元c‐Fos蛋白的表达水平。结果与恐惧消退空白组相比,恐惧消退模型组BLA内的DA、NE含量较高(P<0.01),5‐HT含量显著较低(P<0.01)。与恐惧消退模型组相比,恐惧消退给药组BLA内的DA、NE的含量较低(P<0.05,P<0.01),5‐HT含量较高(P<0.01)。与恐惧消退空白组相比,恐惧消退模型组的BLA内c‐Fos蛋白表达数量较高(P<0.01)。与模型组相比,恐惧消退给药组的BLA内c‐Fos蛋白表达数量较低(P<0.01)。结论朱砂安神丸可以拮抗条件性恐惧,其拮抗作用主要体现在恐惧记忆消退阶段,并可能参与调节BLA区域单胺类神经递质含量以及该区域中c‐Fos蛋白表达水平。

Objective To explore the effect of ZhushaAnshen pill in antagonizing conditioned fear in rats based on the changes of monoamine neurotransmitter and c‐Fos protein expression in basolateral amygdala(BLA).Methods SD rats recovered 3 days after probe implantation.They were randomly divided into 6 groups:fear acquisition blank group,fear acquisition model group,fear acquisition administration group(0.81 g/kg),fear extinction blank group,fear extinction model group and fear regression administration group(0.81 g/kg).The administration group was given ZhushaAnshen pill suspension(0.09 g/mL,9 mL/kg)at 7 am,and the blank group and model group were given the same volume of double distilled water,once a day for 7 consecutive days.After the last administration,the animal models of conditioned fear were reproduced and regressed.The regression test lasted for 7 days,and the drug was given at the same time.The contents of monoamine neurotransmitters dopamine(DA),norepinephrine(NE)and 5‐hydroxytryptamine(5‐HT)in BLA of rats were detected by in vivo microdialysis combined with electrochemical detector(HPLC‐ECD),and the expression level of c‐Fos protein in BLA neurons was determined by immunohistochemistry.Results Compared with the fear extinction model group,the contents of DA and NE in BLA were lower in the fear extinction administration group(P<0.05,P<0.01),but the contents of 5‐HT were higher(P<0.01).Compared with the fear extinction blank group,the expression of c‐Fos protein in BLA was higher in the fear extinction model group(P<0.01).Compared with the fear extinction model group,the expression of c‐Fos protein in BLA was lower in the fear extinction administration group(P<0.01).Conclusion ZhushaAnshen pill can antagonize conditioned fear,and its antagonistic effect is mainly reflected in the fading stage of fear memory,and may be involved in regulating the content of monoamine neurotransmitters and the expression of c‐Fos protein in BLA.