左归丸对围绝经期抑郁症模型小鼠行为学影响及神经保护机制研究

Research of Neuroprotective Mechanism and Effects of Behavior of Zuogui Wan on Perimenopause Depression Model in Mice

目的探讨左归丸对围绝经期抑郁症模型小鼠神经保护作用及机制。方法采用双侧卵巢切除(OVX)联合慢性不可预知性温和应激(CUMS)法制备小鼠围绝经期抑郁症模型,随机分为假手术组、围绝经期抑郁症模型组、氟西汀+雌激素组(3mg/kg+0.15mg/kg)、左归丸低、中、高剂量组(11.5mg/kg、23.0mg/kg、46.0mg/kg),每组10只。观察各组小鼠行为学变化,连续治疗21天后取材。酶联免疫吸附(ELISA)法检测血清雌二醇(E 2)及脑组织单胺类神经递质5-羟色胺(5-HT)、去甲肾上腺素(NE)及多巴胺(DA)含量,免疫印迹(Western blot)法观察海马组织环磷酸腺苷反应元件结合蛋白(CREB)/脑源性神经营养因子(BDNF)信号通路主要蛋白表达。结果与假手术组比较,围绝经期抑郁症模型组小鼠表现抑郁样行为,主要为自主活动及探索活动减少,行为绝望时间增加,血清E 2含量减少,脑组织单胺类神经递质5-HT、NE及DA含量减少,海马组织CREB-1、p-CREB-1、BDNF、酪氨酸蛋白激酶B(TrkB)蛋白表达减少;给予左归丸低、中、高剂量治疗后,可改善围绝经期抑郁症模型小鼠抑郁样行为,表现为提高自主活动及探索活动能力,减少行为绝望程度,增加血清E 2含量,增加脑组织单胺类神经递质5-HT、NE及DA含量,增加海马组织CREB-1、p-CREB-1、BDNF、TrkB蛋白表达;各剂量组组间比较,以中剂量较为明显(P<0.05)。结论左归丸可改善围绝经期抑郁症模型小鼠抑郁样行为,其神经保护机制与增加血清E 2含量、增加脑组织单胺类神经递质含量、上调CREB-BDNF信号通路主要蛋白CREB-1、p-CREB-1、BDNF、TrkB蛋白表达有关。

Objective To investigate the neuroprotective mechanism and the behavior effects of Zuogui Wan on perimenopause depression model in mice.Methods We used OVX+CUMS methods as the perimenopausal depression model in mice.The mice were randomly divided into the sham group,the perimenopausal depression model group,the fluoxetine+estrogen group(3mg/kg+0.15mg/kg),the Zuogui Wan low,middle and high dose groups(11.5mg/kg,23.0mg/kg,46.0mg/kg).The each group contained 10 mice.In this study,the mice were garaged with drug or physiological saline one hour before the daily stress for 21 days.The behavior change of mice were tested by the open filed test(OFT)and the tail suspension test(TST).All of the contents of the estradiol(E 2)in serum,the monoamine neurotransmitter serotonin(5-HT),the norepinephrine(NE)and the dopamine(DA)in brain were observed by ELISA method.The main protein expression of CREB/BDNFsignaling pathways in hippocampus tissue were observed by Western blot method.Results To compared with the sham group,the depression-like behavior of mice were appeared on the perimenopausal depression model group.The specific performance of autonomic activity and exploration activity were decreased in the OFT,the behavioral despair time were increased in the TST,all of the content of E 2 in serum,5-HT,NE and DA in the brain were decreased,the protein expressions of CREB-1,p-CREB-1,BDNF and TrkB in hippocampus tissue were decreased.Compared with other Zuigui Wan dose groups,the middle dose group was more obvious,and the difference was statistically significant(P<0.05).If we gave them FLU+E 2 and the different dose Zuogui Wan,we can improve them.These mice performance for increasing autonomous activities and exploratory behavior in the OFT,decreasing despair time in the TST,increasing serum E 2 levels and the contents of 5-HT,NE and DA in the brain tissue,increasing the protein expression of CREB-1,p-CRE-1,BDNF and TrkB in hippocampus tissue(P<0.05).Conclusion Zuogui Wan can improve depression-like behavior on perimenopausal depression model in mice.The neuron protection mechanisms were concerned about increasing the content of serum E 2,the levels of the monoamine neurotransmitter and the protein expressions of CREB-1,p-CRE-1,BDNF and TrkB.