辛弗林对CUMS大鼠抑郁样行为和血清代谢组学的影响

目的 通过观察分析大鼠慢性应激引起的行为学和血清代谢组变化以及辛弗林抗抑郁的有效治疗,发现潜在的抗抑郁新药先导化合物和生物标志物。方法 将24只大鼠分为正常组、模型组和辛弗林组。模型组为6周慢性温和不可预知应激(CUMS)模型大鼠,用体质量变化、糖水偏好、开场实验作为抑郁评估指标,后2周辛弗林(Syn 20 mg/kg)干预治疗后采集大鼠血清进行气相色谱-质谱联用(GC-MS/MS)测定,研究CUMS大鼠血清内源性代谢物变化,以及辛弗林的干预作用,寻找其抗抑郁潜在的先导化合物和生物标志物,并找到相关靶向代谢通路。结果 辛弗林干预后,在增加CUMS大鼠体质量、提高糖水偏好率、增加开场运动总距离和理毛次数的同时,还能逆转CUMS大鼠血清中磷酸乙醇胺、苯甲酸、胆固醇、谷氨酸等14种代谢物的变化使其趋于正常组,并发现谷氨酰胺和谷氨酸的代谢、缬氨酸和异亮氨酸的生物合成等9条主要牵涉辛弗林的靶向代谢通路。结论 辛弗林可能通过调节谷氨酰胺和谷氨酸的代谢、缬氨酸和异亮氨酸的生物合成等生物学途径发挥对CUMS大鼠抑郁样行为的调正作用。      

慢性应激模型大鼠脑内5-HT1A受体表达分析

目的 探讨慢性不可预见性轻度应激(CUMS)模型大鼠各脑区(海马、中缝核、前额叶及纹状体)5-HT1A受体表达及与旷场行为变化的关系;西酞普兰对5-HT1A受体表达的影响.方法 将24只雄性SD大鼠随机分为3组(n=8):A组为对照组;B组为CUMS应激组;C组为CUMS应激+西酞普兰用药组(每天腹腔注射西酞普兰水溶液2 ml,10mg/kg).实验为期6周,通过旷场试验评价大鼠行为,6周后处死大鼠获取脑组织,采用Real-Time PCR检测各脑区5-HT1A mRNA表达水平,并分析其与矿场行为的相关性.结果 B组海马5-HT1A mRNA表达较A组有增高趋势(P=0.05),其余各脑区5-HT1A mRNA表达均差异无统计学意义(P＞0.05).海马5-HT1AmRNA表达大鼠移动次数、转身次数呈正相关,与总不动时间呈负相关;中缝核、纹状体5-HT1A mR-NA表达与中心移动距离呈正相关,纹状体5-HT1A mRNA表达与中心停留时间呈正相关(P均＜0.05).结论 慢性应激可能会引起海马5-HT1A受体表达增高;抗抑郁剂西酞普兰对5-HT1A受体表达无影响.5-HT1A mRNA表达与旷场行为变化有相关性.     

丙戊酸钠抗抑郁作用涉及改善海马NET和5-HTT表达

采用孤养结合慢性不可预见性温和刺激(CUMS)方式建立抑郁模型,探讨丙戊酸钠抗抑郁作用与改善海马去甲肾上腺素转运体(norepinephrine transporter,NET)、5-羟色胺转运体(serotonin transporter,5-HTT)表达之间的关系。60只雄性SD大鼠随机分为对照组(NG)、模型组(MG)、丙戊酸钠对照组(VNG)及丙戊酸钠模型组(VMG)。通过开场实验、强迫游泳试验评价大鼠抑郁行为。通过生物化学法测定血清丙二醛(MDA)含量及超氧化物酶(SOD)、过氧化氢酶(CAT)活性;以实时定量RT-PCR法及Western blot分析丙戊酸钠对大鼠海马NET、5-HTT mRNA及蛋白表达的影响。实验结果表明:CUMS致模型组大鼠活动减少,不动时间明显增加,MDA含量升高,SOD、CAT活性下降,海马NET mRNA及蛋白表达显著升高,5-HTT mRNA及蛋白表达明显降低,而丙戊酸钠干预可明显阻遏以上变化,但对对照组大鼠无显著影响。以上结果提示,丙戊酸钠抗抑郁作用可能涉及改善CUMS大鼠海马NET及5-HTT mRNA及蛋白的表达。     

Neuropharmacological effect of novel 5-HT3 receptor antagonist,N-n-propyl-3-ethoxyquinoxaline-2-carboxamide (6n) on chronic unpredictable mild stress-induced molecular and cellular response: Behavioural and biochemical evidences

BackgroundChronic unpredictable stressors can produce a situation similar to human depression and such animal models can be used for the preclinical evaluation of antidepressants. The 5-HT₃ receptor antagonists modulate serotonergic pathways and show antidepressant-like effect in various animal models of depression.MethodsIn this study, a novel and potential 5-HT₃ receptor antagonist N-n-propyl-3-ethoxyquinoxaline-2-carboxamide (6n) with good Log P (2.52) value and pA₂ (7.6) values, synthesized in our laboratory was explore to study the effects on CUMS-induced behavioural and biochemical alterations in mice. Mice were subjected to different stress paradigms daily for a period of 28 days to induce depressive-like behaviour.ResultsCUMS caused depression-like behaviour in mice, as indicated by the significant decrease in sucrose consumption and increase in immobility time in the forced swim test (FST) while there was no significant effect on spontaneous locomotor activity (SLA) observed. In addition it was found that lipid peroxide and nitrite levels were significantly increased, whereas glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) levels were decreased in brain tissue of CUMS-treated mice. Compound 6n (1 and 2 mg/kg, po, 21 days) and fluoxetine treatment (20 mg/kg, po, 21 days) significantly altered the CUMS-induced behavioural (increased immobility period, reduced sucrose preference) and biochemical (increased lipid peroxidation, increased brain nitrite; decreased GSH, SOD and CAT levels) parameters while there was no significant effect of observed on SLA.ConclusionCompound 6n produced antidepressant-like effects in behavioural despair paradigm in chronically stressed mice by restoring antioxidant enzyme activity up to significant level.     

蜘蛛香环烯醚萜部位干预CUMS致抑郁小鼠脑组织的1H-NMR代谢组学分析

目的:利用代谢组学技术分析慢性温和不可预知应激(CUMS)抑郁模型小鼠脑组织样本,寻找与抑郁相关的差异代谢物,探讨蜘蛛香环烯醚萜部位(IEFV)可能的抗抑郁作用机制。方法:42只昆明小鼠随机分为6组,包括正常组,模型组,氟西汀组(2.5 mg·kg-1),IEFV低、中、高剂量组(给药剂量分别为5.73,11.47,22.94 mg·kg-1)。采用CUMS对小鼠造模,以IEFV及阳性药(氟西汀)为干预药物,用行为指标和生化指标进行药效学评价。采用核磁共振氢谱(1H-NMR)代谢组学技术分析IEFV对CUMS抑郁模型小鼠脑组织中内源性物质的影响,结合多变量统计分析方法来确认差异代谢物,并对差异代谢物参与的代谢通路进行富集。结果:造模后,小鼠的不动时间大幅度提高、蔗糖偏好率明显下降,兴奋性神经递质5-羟色胺和去甲肾上腺素显著下降,表明造模成功。给予IEFV和氟西汀后,小鼠不动时间、蔗糖偏好率和兴奋性神经递质向正常水平回调,提示给药后抑郁状态得到了一定程度的缓解。脑组织中内源性代谢物主成分分析(PCA)结果显示,模型组可与正常组明显分开,同时IEFV低、中、高剂量组和氟西汀组均显示有偏离模型组向正常组靠拢的趋势,与行为学结果一致。模型组与正常组进行正交偏最小二乘法-判别分析(OPLS-DA)得到了16个变化显著的差异代谢物,包括12个水溶性差异代谢物和4个脂溶性差异代谢物。通过MetPA数据库分析得到7条潜在靶标代谢通路,包括三羧酸循环(TCA),牛磺酸和亚牛磺酸的代谢,丙氨酸、天冬氨酸和谷氨酸代谢等。IEFV高剂量组可显著回调11种差异代谢物。结论:IEFV可能主要通过影响能量代谢,氨基酸代谢和神经递质水平发挥抗抑郁作用,可为IEFV抗抑郁作用机制的深入研究提供参考依据。     

Polyunsaturated fatty acid supplement alleviates depression-incident cognitive dysfunction by protecting the cerebrovascular and glymphatic systems

IntroductionDepression, the most prevalent mood disorder, has high comorbidity with cerebrovascular disease and cognitive decline. However, there is little understanding of the cellular mechanisms involved in depression and its comorbid cerebrovascular damage and cognition impairment. Here, we tested the prediction that the chronic unpredictable mild stress (CUMS) mouse model would manifest in disturbed glymphatic function and that dietary supplementation with polyunsaturated fatty acids (PUFA) could ameliorate these deficits while alleviating the depression-associated cognitive decline.MethodsTo test the treatment effects of PUFA or Es on behaviours, we applied the tail suspension, open field, and sucrose preference tests to assess depressive symptoms, and applied the Morris water maze test to assess cognition in groups of control, chronic unpredictable mild stress (CUMS), PUFA, and escitalopram (Es) treatment. We measured the extracellular concentrations of dopamine (DA), 5-hydroxytryptamine (5-HT) and noradrenaline (NA) in microdialysates from prefrontal cortex (PFC) by liquid chromatography mass spectrometry. Glia cells and inflammatory factors were analysed with fluorescent immunochemistry and western blot, respectively. We tested brain vasomotor function with two-photon and laser speckle imaging in vivo, and measured glymphatic system function by two-photon imaging in vivo and fluorescence tracer imaging ex vivo, using awake and anesthetized mice. Besides, we monitored cortical spreading depression by laser speckle imaging system. AQP4 depolarization is analysed by fluorescent immunochemistry and western blot.ResultsWe confirmed that CUMS elicited depression-like and amnestic symptoms, accompanied by decreased monoamines neurotransmitter concentration in PFC and upregulated neuroinflammation markers. Moreover, CUMS mice showed reduced arterial pulsation and compliance in brain, and exhibited depolarized expression of AQP4, thus indicating glymphatic dysfunction both in awake and anesthetized states. PUFA supplementation rescued depression-like behaviours of CUMS mice, reduced neuroinflammation and cerebrovascular dysfunction, ultimately improved cognitive performance, all of which accompanied by restoring glymphatic system function. In contrast, Es treatment alleviated only the depression-like behavioural symptoms, while showing no effects on glymphatic function and depression-incident cognitive deficits.ConclusionsThe CUMS depression model entails suppression of the glymphatic system. PUFA supplementation rescued most behavioural signs of depression and the associated cognitive dysfunction by restoring the underlying glymphatic system disruption and protecting cerebral vascular function.     

下丘脑NK2受体在应激介导抑郁症发生中的作用

目的 阐明神经激肽A的受体NK2在慢性不可预见性温和刺激(CUMS)所致的抑郁样行为发生、发展中的可能作用和机制.方法 SD大鼠随机分为对照组、氟西汀组和抑郁模型组3组,行为学检测后,氟西汀组和抑郁模型组均给予孤养+CUMS造成大鼠抑郁症模型,再次行为学检测后,氟西汀组大鼠给予氟西汀腹腔注射21d,抑郁模型组给予同体积生理盐水腹腔注射21d,再次进行行为学检测,麻醉后取大鼠下丘脑,提取组织mRNA和蛋白后,采用荧光定量PCR技术和Western Blot技术检测NK2的表达.结果 应激前3组大鼠的糖水偏好率和强迫游泳的不动时间均无统计学意义.应激后,氟西汀组和抑郁模型组大鼠的糖水摄入明显减少、强迫游泳不动时间明显增长,差异均有统计学意义(P＜0.05).而给予氟西汀后,氟西汀组大鼠的糖水偏好率有所增加,强迫游泳的不动时间有所减少,与对照组比较差异无统计学意义(P＞0.05),但与抑郁组大鼠相比,差异均有统计学意义(P＜0.05).与对照组和氟西汀组比较,抑郁组下丘脑NK2受体的mRNA和蛋白的表达量明显升高,差异有统计学意义(P＜0.05),而氟西汀组与对照组相比差异无统计学意义(P＞0.05).结论 CUMS可引起大鼠行为学改变,造成大鼠抑郁模型;NK2受体的mRNA和蛋白的表达量在应激后大鼠的下丘脑中明显升高,经氟西汀干预后可恢复到正常水平,说明NK2受体与抑郁症的发生、发展过程具有相关性,在抑郁症的发病机制中可能发挥重要作用.     

Notch1 signaling,hippocampal neurogenesis and behavioral responses to chronic unpredicted mild stress in adult ischemic rats

Accumulating evidence indicates that the Notch signaling pathway fulfills important roles in ischemia-stimulated neurogenesis, which may be regarded as an etiological factor in post-stroke depression. Here we explored Notch₁ signaling, hippocampal neurogenesis and behavioral responses to chronic unpredicted mild stress (CUMS) in adult ischemic rats. Animals were treated with permanent middle cerebral artery occlusion followed by an 18 day CUMS procedure. Proliferating cells in the hippocampus and their cell fate were investigated on days 19 and 28 after ischemic surgery. Additionally, expression of the Notch₁ intracellular domain (NICD) and its downstream targets Hes1 and Hes5 was examined. A sucrose preference test and forced swim test were used to assess behavioral responses. CUMS produced depressive-like behaviors and decreased the number of proliferating cells on day 19 (both p < 0.001), accompanied by a decreased expression of both Hes1 and Hes5 in the hippocampus of ischemic animals (p < 0.001). On day 28, CUMS resulted in a decreased number of neurogenically-differentiating cells in the subgranular zone (p < 0.001) while permitting differentiation into astrocytes in the hilus (p < 0.05). Hes1 and Hes5 protein expression levels were increased. The expression of the NICD was significantly decreased at both time-points. CUMS led to expression changes in the Notch₁ signaling cascade in ischemic rats, most of which concerned hippocampal neurogenesis. This suggests that variation in Notch₁ activity and subsequent expression of its downstream targets, including Hes1 and Hes5, may, at least in part, contribute to modulation of ischemia-related hippocampal neurogenesis by CUMS.     

支气管哮喘伴发抑郁大鼠模型的建立

目的 探讨建立支气管哮喘伴发抑郁动物模型的方法 .方法 选择Wistar大鼠,随机分为对照组与模型组,模型组采用卵蛋白(ovalbumin,OVA)激发法建立哮喘模型,在此基础上给予慢性轻度不可预见性应激(chronic unpredictable mild stress,CUMS)28 d,观察大鼠体质量、体征、肺组织结构、支气管肺泡灌洗液白细胞计数等变化,并用Open-field实验评价大鼠活动度和好奇心,通过糖水消耗实验评价大鼠快感缺乏与否.结果 与对照组相比,模型组大鼠体质量增长明显缓慢(P<0.05);肺组织呈哮喘样病理改变;支气管肺泡灌洗液白细胞总数、嗜酸粒细胞及淋巴细胞增多;Open-field实验,大鼠垂直活动得分、水平活动得分显著降低(P<0.05);糖水消耗量明显减少(P<0.05).结论 OVA激发复合CUMS可成功制备支气管哮喘伴发抑郁大鼠模型.