醒脑解郁方对卒中后抑郁模型大鼠行为学影响及作用机制

Effect of Xingnao Jieyu recipe on behavior of rats with post-stroke depression and its mechanism

目的探讨醒脑解郁方对卒中后抑郁(PSD)模型大鼠行为学的影响及机制。方法将72只健康成年雄性SD大鼠随机分为6组:空白组(Con组)、卒中组(MCAO组)、卒中后抑郁组(PSD组)、醒脑解郁低剂量组(XNJY-L)、醒脑解郁高剂量组(XNJY-H)、氟西汀组(Flu组)。除Con组外,其余组均采用MCAO法制备卒中大鼠模型;造模后,用Longa 5分法对大鼠神经功能进行评定,选取评分1~3分的大鼠进行后续实验,除Con组和MCAO组外,其余组造模成功大鼠均使用慢性不可预知温和刺激的方法,持续21 d制备抑郁模型。在卒中模型制备成功后的第2天,各给药组开始接受相应药物的干预,持续21 d。观察大鼠体质量、行为学状态;采集大鼠海马标本,HE染色法观察大鼠海马组织病理形态变化;ELISA法检测血清IL-6、IL-10;Western blotting法检测大鼠海马Keap1、核因子E2相关因子2(Nrf2)、血红素氧化酶(HO-1)蛋白表达。结果与Con组比较,PSD组体质量、强迫游泳测试时间、糖水消耗量、水平及垂直运动得分低于Con组(P均<0.05);PSD组海马组织神经元数目减少、稀疏,细胞间空隙增宽,胞体回缩、溶解,胞质胞核甚至空洞、消失;PSD组海马Keap1蛋白表达上调,海马Nrf2、HO-1蛋白表达下调(P均<0.05);血清IL-6、IL-10水平下降(P均<0.05);与PSD组相比,XNJY-L组、XNJY-H组及Flu组体质量、强迫游泳时间、糖水消耗量、水平及垂直运动得分均高(P均<0.05);海马组织神经细胞结构大部分保持完好,受损程度较轻,细胞排列紧密,尼氏小体数目丰富,胞核、胞质等结构存在完好;海马Nrf2、HO-1蛋白表达上调(P均<0.05);血清IL-6、IL-10水平升高(P均<0.05)。结论醒脑解郁方能改善PSD大鼠的抑郁样行为学症状,发挥抗抑郁作用,其机制可能与抑制海马神经组织氧化应激、增强Nfr2/HO-1信号通路活性、增强抗氧化应激能力有关。

Objective To investigate the effect of Xingnao Jieyu recipe(XNJY)on behavior of rats with post-stroke depression(PSD)and the mechanism of action.Methods Seventy-two male SD rats were randomly divided into six groups:control(Con),stroke(MCAO),PSD,low-dose XNJY(XNJY-L),high-dose XNJY(XNJY-H),and fluoxetine(Flu)groups.We established the stroke models using the MCAO method in all groups except the Con group.Neurological function was evaluated using the Longa 5-point scale,and we included only rats scoring 1–3 in subsequent experiments.We created PSD models in all groups except Con and MCAO by subjecting the rats to 21 days of chronic unpredictable mild stress(CUMS).Drug interventions began on the second day after stroke modeling and continued for 21 days.We assessed body weight,behavior,and hippocampal pathology through HE staining.Serum IL-6 and IL-10 levels were measured using ELISA,and Western blotting was employed to detect the expression levels of hippocampal Keap1,nuclear factor E2-associated factor 2(Nrf2),and heme oxygenase(HO-1)protein.Results Compared with the Con group,the PSD group had lower body weight,shorter forced swimming time,lower sucrose consumption,and lower scores of horizontal and vertical movements(all P<0.05);in the PSD group,the number of neurons in the hippocampal tissues decreased and became sparse,the intercellular space widened,the cell bodies shrank and dissolved,and the cytoplasm and cell nuclei even became hollow and disappeared;Keap1 protein expression was up-regulated,while Nrf2 and HO-1 levels decreased,and se rum IL-6 and IL-10 levels decreased(all P<0.05).Compared with the PSD group,the XNJY-L,XNJY-H,and Flu groups had higher body weight,longer forced swimming time,higher sucrose consumption,and higher scores of horizontal and vertical movements(all P<0.05).The structure of nerve cells in the hippocampal tissues remained mostly intact with a relatively minor degree of damage;the cells were closely arranged,and there was an abundance of Nissl bodies;the structures such as cell nuclei and cytoplasm remained intact;the protein expression levels of Nrf2 and HO-1 in the hippocampus were up-regulated and the levels of IL-6 and IL-10 in the serum increased(all P<0.05).Conclusions Xingnao Jieyu recipe improves depressive-like behavioral symptoms in PSD rats,exerting an anti-depressant effect to achieve the treatment goal for PSD.Its mechanism may be related to the inhibition of oxidative stress in hippocampal neural tissues,activation of the Nrf2/HO-1 signaling pathway,and enhancement of antioxidant capacity.