肉蔻五味丸抗抑郁作用及机制研究

Antidepressant effect and mechanism of Roukou wuwei pills

目的:探讨肉蔻五味丸的抗抑郁作用及其作用机制,为临床用药提供依据。方法:雄性C57BL/6小鼠随机分为空白对照组(control)、模型组(model)、盐酸氟西汀(fluoxetine 10 mg/kg)阳性药组、肉蔻五味丸低剂量(rk 0.2 g/kg)组、肉蔻五味丸高剂量(rk0.5 g/kg)组,给予慢性轻度不可预见性加孤养刺激建立抑郁模型,建模同时灌胃给予小鼠药物治疗。给药五周后,进行行为学检测(强迫游泳实验、悬尾实验和高架十字迷宫实验);LC-MS/MS法检测海马内5-羟色胺(5-HT)等神经递质含量;q PCR法检测海马内5-羟色胺1A受体(5-HT1AR)、5-羟色胺7受体(5-HT7R)、GABAA型受体(GABAAR)、GABAB型受体(GABABR)、神经生长因子(NGF)和胶质细胞源神经营养因子(GDNF)基因表达;ELISA法检测小鼠血清内促肾上腺皮质激素(ACTH)和皮质酮(COR)的含量。结果:低剂量肉蔻五味丸有效减少慢性应激导致的小鼠游泳和悬尾不动时间的增加(P<0.05),降低模型组小鼠进入高架中央区活动时间(P<0.05),逆转小鼠海马5-HT(P<0.001)、5-HT/5-HIAA(P<0.001)的降低以及GABABR(P<0.01)、NGF(P<0.01)、GDNF(P<0.001)基因表达的降低(P<0.05),并且降低血清中ACTH的水平(P<0.05)。结论:肉蔻五味丸低剂量具有抗抑郁作用,其机制可能与提高抑郁小鼠海马5-HT含量、GABABR、NGF和GDNF基因表达以及降低血清中ACTH的水平有关。

AIM：To explore the antidepressant effect of Roukouwuweiwan （rk） and its action mechanism,providing the theoretical basis for clinical medication.METHODS：Male C57BL/6 mice were randomly divided into control group,model group,fluoxetine group （10 mg/kg）,rk low-dose group （rk 0.2 g/kg）,rk high-dose group （rk 0.5 g/kg）.Each group of mice was single caged and given unpredicted chronic mild stress （UCMS） except the control group.Drugs were administered intragastrically from the first day of stimulation and lasted for five weeks.Behavioral changes of mice were evaluated by forced swimming test （FST）,tail suspension test （TST） and elevated plus maze test （EPMT）.Hippocampal neurotransmitters were examined using LC-MS/MS method.mRNA expressions of depression related genes such as 5-HT1 AR,5-HT7R,GABAAR,GABABR,NGF,and GDNF in hippocampus were detected by qPCR method.ACRH and COR level in mouse serum were detected by ELISA method.RESULTS：The immobility time in FST and TST （P 〈 0.05） and time in middle field of EPMT （P 〈 0.05） was reduced in rk （0.2 g/kg） treated UCMS mice.The content of 5-HT （P 〈0.001） as well as the ratio of 5-HT to 5-HIAA （P 〈 0.001） in hippocampus of UCMS mice were increased.Also,mRNA expressions of hippocampal GABABR （P 〈 0.01）,NGF （P 〈 0.01） and GDNF （P 〈 0.001） were increased with rk （0.2 g/kg） while that of ACTH （P〈0.05） in serum was reduced.CONCLUSION：Low-dose rk showed better antidepressant effect,which might be mediated by elevation of hippocampal 5-HT content,up-regulation of hippocampal GABABR,NGF and GDNF mRNA expression,and down-regulation of ACTH in serum.