目的:探讨柴胡疏肝散(CSS)对雌性小鼠的快速抗抑郁潜力及其作用机制。
方法:液相色谱质谱(LC-MS)/MS用于测定CSS中主要成分的含量,以确定其稳定性。雌性C57BL/6J小鼠随机分为4组,包括对照(盐水),车辆(盐水),CSS(4μg/kg)和氯胺酮(30mg/kg)组。对小鼠进行4周的不规则应激刺激,建立慢性轻度应激(CMS)模型,然后接受了一次药物治疗。两小时后,进行了行为测试,包括露天测试,尾部悬挂试验(TST),强迫游泳试验(FST)新奇抑制喂养试验(NSF),和蔗糖偏好试验(SPT)。Westernblot分析用于检测N-甲基-D-天冬氨酸受体(NMDA)亚型[N-甲基-D-天冬氨酸受体1(NR1)的表达水平,NR2A,NR2B],突触蛋白[突触素1和突触后密度蛋白95(PSD95)],脑源性神经营养因子(BDNF)。此外,CSS的快速抗抑郁作用是通过激活谷氨酸受体的药理学技术和光遗传学干预来测试的,NMDA.
结果:与载体组相比,一次使用CSS(4g/kg)可以逆转TST中的所有行为缺陷,FST,由CMS引起的SPT和NSF(P<0.05或P<0.01)。CSS也显著降低了NMDA亚型的表达(NR1,NR2A,NR2B)在小鼠海马2h(均P<0.01)。此外,类似于氯胺酮,CSS增加了突触蛋白和BDNF的水平(P<0.05或P<0.01)。此外,CSS的快速抗抑郁作用被海马NMDA受体的瞬时激活所阻断(均P<0.01)。
结论:CSS通过改善雌性CMS小鼠行为缺陷的快速抗抑郁作用取决于NMDA受体的快速抑制和突触蛋白的激活。
OBJECTIVE: To explore the rapid antidepressant potential and the underlying mechanism of Chaihu Shugan San (CSS) in female mice.
METHODS: Liquid chromatography mass spectrometry (LC-MS)/MS was used to determine the content of main components in CSS to determine its stability. Female C57BL/6J mice were randomly divided into 4 groups, including control (saline), vehicle (saline), CSS (4 g/kg) and ketamine (30 mg/kg) groups. Mice were subjected to irregular stress stimulation for 4 weeks to establish the chronic mild stress (CMS) model, then received a single administration of drugs. Two hours later, the behavioral tests were performed, including open field test, tail suspension test (TST), forced swimming test (FST), novelty suppression feeding test (NSF), and sucrose preference test (SPT). Western blot analysis was used to detect the expression levels of N-methyl-D-aspartate receptor (NMDA) subtypes [N-methyl-D-aspartate receptor 1 (NR1), NR2A, NR2B], synaptic proteins [synapsin1 and post synaptic density protein 95 (PSD95)], and brain-derived neurotrophic factor (BDNF). Moreover, the rapid antidepressant effect of CSS was tested by pharmacological technologies and optogenetic interventions that activated glutamate receptors, NMDA.
RESULTS: Compared with the vehicle group, a single administration of CSS (4 g/kg) reversed all behavioral defects in TST, FST, SPT and NSF caused by CMS (P<0.05 or P<0.01). CSS also significantly decreased the expressions of NMDA subtypes (NR1, NR2A, NR2B) at 2 h in hippocampus of mice (all P<0.01). In addition, similar to ketamine, CSS increased levels of synaptic proteins and BDNF (P<0.05 or P<0.01). Furthermore, the rapid antidepressant effects of CSS were blocked by transient activation of NMDA receptors in the hippocampus (all P<0.01).
CONCLUSIONS: Rapid antidepressant effects of CSS by improving behavioral deficits in female CMS mice depended on rapid suppression of NMDA receptors and activation of synaptic proteins.