天王补心丸对慢性不可预知应激抑郁模型小鼠行为学,HPA轴,海马GSK3β磷酸化及BDNF表达的影响

Effect of Tianwang Buxin Pills on Behavior, HPA Axis, BDNF Expression and Phosphorylation of GSK3β in Mice with Chronic Unpredictable Stress

目的:研究天王补心丸对慢性应激抑郁模型小鼠行为学、下丘脑-垂体-肾上腺轴(HPA轴),海马糖原合酶激酶3β(GSK3β)磷酸化及脑源性神经营养因子(BDNF)表达的影响,初步探讨其抗抑郁作用机制。方法:将60只雄性ICR小鼠按照糖水偏嗜度随机分为正常组、慢性应激模型组、盐酸氟西汀组(10 mg·kg-1)以及天王补心丸高、中、低剂量组(3.6,1.8,0.9 g·kg-1)。采用慢性不可预知应激实验(CUS)建立抑郁症小鼠模型,各给药组灌胃给药28 d后通过糖水偏好实验、开场行为实验、新奇抑制摄食实验检测其行为学变化。之后分别取小鼠血、肾上腺及海马组织,采用酶联免疫吸附测定(ELISA)法检测小鼠血清中促肾上腺皮质激素(ACTH)和皮质酮(CORT)含量,蛋白免疫印迹法(Western blot)检测海马组织中GSK3β磷酸化及BDNF表达的变化,并计算肾上腺指数。结果:与正常组比较,模型组小鼠的蔗糖水偏嗜度显著降低(P<0.01),开场活动次数明显减少(P<0.05),新奇抑制摄食潜伏期延长(P<0.01),血清ACTH,CORT含量民明显升高(P<0.05,P<0.01),海马组织中GSK3β磷酸化及BDNF表达显著降低(P<0.01),肾上腺指数显著升高(P<0.01);与模型组比较,天王补心丸可以逆转抑郁症模型小鼠行为学变化(P<0.05,P<0.01),显著降低抑郁症模型小鼠血浆ACTH和CORT的含量(P<0.01)和肾上腺指数(P<0.01),同时增加小鼠海马GSK3β磷酸化及BDNF表达水平(P<0.05,P<0.01),其作用效果与盐酸氟西汀相当。结论:天王补心丸对慢性不可预知应激抑郁模型小鼠具有显著的抗抑郁作用,其作用机制与抑制HPA轴活性、上调海马GSK3β磷酸化及BDNF的蛋白表达有关。

Objective: To investigate the effect of Tianwang Buxin pills on behavior,hypothalamus pituitary adrenal axis(HPA axis),hippocampal glycogen synthase kinase 3β(GSK3β)phosphorylation and brain-derived neurotrophic factor(BDNF)expression in mice with chronic unpredictable stress,and explore its mechanisms for antidepressant-like action. Method: Totally 60 male ICR mice were randomly divided into normal group,chronic stress model group,fluoxetine hydrochloride group(10 mg·kg-1)and Tianwang Buxin pills high, middle and low dose groups(3.6, 1.8, 0.9 g·kg-1). The mice were subjected to the chronic unpredictable stress(CUS)protocol for a period of 28 d to induce depressive-like behavior. Then,a sucrose preference test,open-field test and novelty-suppressed feeding test were performed to detect the behavior changes. The blood,adrenal gland and hippocampus of mice were collected. The contents of adrenocorticotropic hormone(ACTH)and corticosterone(CORT)in serum were detected by enzyme-linked immunosorbent assay(ELISA). The changes of GSK3β phosphorylation and BDNF expression in hippocampus were detected by Western blot,and the adrenal index was then calculated. Result: As compared with the normal group,the sucrose water preference was significantly decreased(P<0.01), the number of opening activities was significantly reduced(P<0.05),the feeding latency of novelty inhibition was prolonged(P<0.01),the serum ACTH and CORT contents were significantly increased(P<0.05,P<0.01),GSK3β phosphorylation and BDNF expression levels in hippocampus were significantly decreased(P<0.01),and adrenal index was significantly increased in model group(P<0.01). As compared with the model group,Tianwang Buxin pills treatment significantly reversed the CUS-induced behavioral abnormalities in depression model mice(P<0.05,P<0.01),significantly decreased the levels of plasma ACTH and CORT(P<0.01)and adrenal and adrenal gland index(P<0.01),while increased GSK3β phosphorylation and BDNF expression in hippocampus(P<0.05,P<0.01),with its effect similar to that of fluoxetine hydrochloride. Conclusion:Tianwang Buxin pills produced antidepressantlike effects in chronic unpredictable stress mice,and its mechanism may be associated with inhibiting HPA axis activity and up-regulating GSK3β phosphorylation and BDNF protein expression in hippocampus.