中药复方开心散调控神经营养因子抗抑郁物质基础与作用机制研究

目的：观察开心散调控神经营养因子抗抑郁的物质基础与作用机制。方法：制备开心散大孔树脂乙醇洗脱部位,采用悬尾和强迫游泳评价抗抑郁效果。采用ELISA法测定海马中神经生长因子（NGF）与脑源性神经营养因子（BDNF）的含量。利用大鼠星型胶质瘤C6细胞系和大鼠肾上腺嗜铬瘤PC12细胞系,评价不同乙醇洗脱部位促进NGF与BDNF表达和促进PC12细胞分化的作用。结果：开心散各效用部位均能显著缩短小鼠悬尾与强迫游泳不动时间,其中70%乙醇洗脱部位抗抑郁效用最强,该部位还能通过cAMP信号通路上调C6细胞中NGF与BDNF的表达。10%乙醇洗脱部位促PC12细胞分化的能力最强。人参皂苷是促进神经营养因子表达的主要效用成分。结论：开心散调控神经营养因子是其抗抑郁效用的重要作用机制。     

中药复方开心散调控慢性压力应激小鼠海马炎性细胞因子水平抗抑郁作用机制研究＊

目的 探究中药复方开心散调控中枢神经炎症系统抗抑郁的作用机制。方法 制备开心散不同配伍比例提取物,采用小鼠慢性应激压力（CUMS）模型,通过糖水偏嗜实验、悬尾实验和强迫游泳实验评价开心散不同配伍比例的抗抑郁功效。采用ELISA法测定受试动物海马组织促炎物质如脂多糖（LPS）、IL-1β、IL-6、TNF-α的含量。采用免疫组织化学染色法观察受试动物海马区小胶质细胞形态。进而采用LPS促进小鼠小胶质永生化细胞（BV2）炎性因子表达细胞模型,采用ELISA法评价开心散抑制炎症因子表达的作用,采用Western blotting法考察NF-κB信号通路的调控情况。结果 开心散不同配伍比例能显著上调模型小鼠糖水偏嗜率,缩短小鼠悬尾和强迫游泳不动时间,以D-652效用最强。该配伍比例通过抑制NF-κB的入核和抑制NF-κB磷酸化等环节,下调BV2细胞中炎症因子IL-1β、IL-6、TNF-α的表达水平。结论 对中枢神经炎症的调控是中药复方开心散抗抑郁的重要作用环节。     

陈皮提取物改善慢性温和不可预知应激小鼠行为和海马BDNF的研究

目的:探讨陈皮提取物对慢性温和不可预知应激(CUMS)抑郁模型小鼠的行为和对脑源性神经营养因子(BDNF)表达水平的影响。方法:ICR小鼠根据糖水偏好百分比随机分为空白对照组、模型对照组、阳性对照盐酸氟西汀20 mg·kg-1组及陈皮提取物100,200,400 mg·kg-1组;除空白对照组外,各组均接受CUMS,造模28 d后开始灌胃给药;各组连续给药28 d后,糖水偏好实验测定糖水偏好百分比、新奇抑制摄食实验测定摄食潜伏期、强迫游泳实验测定不动时间;荧光实时定量PCR和ELISA法测定大脑海马BDNF含量。结果:慢性应激后,模型对照组小鼠糖水偏好百分比减少,摄食潜伏期延长,不动时间增多,海马BDNF含量显著降低;与模型对照组比较,陈皮提取物组能显著改善抑郁小鼠行为学指标,显著提高小鼠海马BDNF含量。结论:陈皮提取物对CUMS模型小鼠有抗抑郁作用,其机制可能与增加海马BDNF有关。     

逍遥丸对皮质酮诱导小鼠抑郁样行为的干预机制※

目的:观察逍遥丸对皮质酮诱导小鼠抑郁样行为的干预作用,并探讨其分子机制。方法:将50只ICR雄性小鼠,随机分为5组:正常组、皮质酮模型组、阳性对照氟西汀组(20 mg/kg)、低剂量逍遥丸(200 mg/kg)组、高剂量逍遥丸组(600 mg/kg),通过皮下注射皮质酮诱导小鼠抑郁模型。持续35天后,采用糖水偏好实验和强迫游泳实验评价动物抑郁样行为;采用ELISA方法测小鼠血清中皮质酮含量及小鼠海马组织中脑源性神经营养因子(BDNF)的含量。结果:皮质酮可以降低糖水偏好值、增加小鼠强迫游泳的不动时间,而逍遥丸可以显著提高糖水偏好值、减少小鼠不动时间;长期注射皮质酮可增加血清皮质酮水平,降低海马组织中BDNF含量,而逍遥丸可以降低小鼠血清中皮质酮的含量并且能够提高海马组织中BDNF含量。结论:逍遥丸可以有效降低小鼠血清中皮质酮的含量并增加小鼠海马中BDNF含量,改善神经营养系统,产生抗抑郁样作用。     

舒郁散对慢性应激抑郁大鼠行为及海马BDNF表达的影响

目的观察中药舒郁散对慢性应激抑郁大鼠的行为学变化和海马脑源性神经营养因子(BDNF)蛋白表达的影响。方法 50只SD大鼠随机分为正常对照组、模型组、百忧解组、舒郁散小剂量组、舒郁散大剂量组;对大鼠进行21 d的应激刺激建立慢性应激抑郁大鼠模型,观察各组糖水摄入量及强迫游泳测试等行为学指标变化,用免疫组化方法检测大鼠BDNF蛋白表达。结果造模后,模型组大鼠糖水消耗明显下降;大剂量舒郁散能明显增加慢性应激抑郁大鼠的糖水消耗量,能明显缩短抑郁大鼠游泳不动时间,与模型组比较有显著差异;模型组大鼠海马BDNF蛋白表达减少,大剂量舒郁散给药后海马BDNF蛋白表达阳性细胞明显增加,且较模型组明显。结论舒郁散可改善慢性应激抑郁大鼠行为、具有抗抑郁的作用,其抗抑郁作用可能与增加海马BDNF蛋白表达有关。     

人参果胶酸性组分调控BDNF/TrkB信号通路抗抑郁样行为的研究

目的探讨了人参果胶酸性组分调控BDNF/TrkB信号通路抗抑郁样行为。方法强饲给予小鼠人参果胶酸性组分12天,通过开场实验,高架十字迷宫实验,社交干扰实验及强迫游泳实验评价其抗抑郁样行为,Western Blotting检测小鼠海马组织脑源性神经营养因子BDNF及其受体TrkB的蛋白表达。结果人参果胶酸性组分具有抗抑郁样活性,该表观有效性伴随着小鼠海马组织BDNF及其受体TrkB的蛋白表达增加。结论人参果胶酸性组分通过诱导小鼠海马BDNF及其受体TrkB的蛋白表达上调进而增加神经元的存活,影响神经元突触的可塑性发挥抗抑郁的作用。     

基于BDNF 通路探讨樱桃叶水煎液对CUMS 大鼠的抗抑郁作用机制

目的探讨樱桃叶水煎液对慢性不可预知性温和应激（CUMS）模型大鼠的抗抑郁作用及可能机制。 方法将雄性Sprague-Dawley（SD）大鼠随机分为空白组、CUMS 模型组、盐酸氟西汀组（2 mg·kg-1）以及樱桃叶 水煎液低、中、高剂量组（1.8、2.7、3.6 g·kg-1）。复制CUMS 模型,采用强迫游泳实验、悬尾实验和糖水偏好 实验观察大鼠行为学变化;酶联免疫吸附实验（ELISA）测定大鼠血清促肾上腺皮质激素（ACTH）含量;Western Blot 法测定大鼠海马组织中脑源性神经营养因子（BDNF）、酪氨酸激酶受体B（TrkB）、Ras 相关C3 肉毒杆菌毒 素底物1（Rac1）、LIM 激酶1（LIMK1）、丝切蛋白（cofilin）的表达水平。结果①与空白组比较,CUMS 模型组 大鼠强迫游泳实验和悬尾实验的不动时间增加（P＜0.01）,糖水偏好系数下降（P＜0.01）;血清ACTH 水平上调 （P＜0.01）;海马组织BDNF、TrkB、Rac1、LIMK1、cofilin 蛋白表达降低（P＜0.01）。②与CUMS 模型组比 较,樱桃叶水煎液干预后大鼠强迫游泳实验和悬尾实验的不动时间减少（P＜0.05,P＜0.01）,而糖水偏好系数 上升（P＜0.01）;血清ACTH 水平下降（P＜0.01）;海马组织BDNF、TrkB、Rac1、LIMK1、cofilin 蛋白表达增 强（P＜0.05,P＜0.01）。结论樱桃叶水煎液具有一定的抗抑郁作用,其机制可能与调节BDNF/TrkB 及其下游 的Rac1/LIMK1/cofilin 信号通路有关。     

舒郁宁心方对慢性应激抑郁模型大鼠行为学及海马BDNF、TrkB表达的影响

目的 观察舒郁宁心方对慢性应激抑郁模型大鼠行为学以及海马脑源性神经营养因子（brain derived neurotrophic factor,BDNF） 及其主要受体酪氨酸激酶B（tyrosine kinase B,TrkB）表达的影响,探讨其抗抑郁机制。方法 将60只成年SD大鼠随机分为正常对照组、模型组、西药组及中药大、中、小剂量组,每组10只。对大鼠进行21天的应激刺激建立慢性应激抑郁模型。除正常对照组外,其他5组大鼠在造模第22天开始灌胃,模型组灌胃生理盐水,中药大、中、小剂量组分别给予25.0、7.5、2.5 g/kg舒郁宁心方灌胃;西药组给予盐酸氟西汀混悬液（12 mg/kg）灌胃;均连续给药3周。分别于0（造模前）、3周（造模成功后）、6周（给药结束后）测定每只大鼠体重;敞箱实验检测大鼠行为学;强迫游泳实验记录大鼠5 min内不动时间。实验结束后脑区取材,测定大鼠脑组织中BDNF及TrkB表达水平。结果 与正常对照组比较,模型组大鼠体重增长缓慢,行为学指标下降,强迫游泳不动时间延长,海马BDNF及TrkB表达减弱,差异均有统计学意义（P<0.05,P<0.01）。给药结束后,中药大剂量组和西药组大鼠体重增加,大鼠行为学改善,强迫游泳不动时间缩短,海马BDNF、TrkB表达增强,与模型组比较,差异均有统计学意义（P<0.05,P<0.01）。结论 舒郁宁心方可能通过改善脑组织BDNF和TrkB的表达水平发挥抗抑郁作用。     

豆腐果苷对慢性应激小鼠海马神经元再生的影响

 目的 研究豆腐果苷对慢性应激小鼠海马神经元再生的影响,探讨其抗抑郁作用的可能机制。方法 给予小鼠多种慢性不可预知性应激刺激,建立小鼠抑郁模型,刺激前30 min给予豆腐果苷（10、20、40 mg·kg^-1,ig）,连续28 d。用免疫组化实验检测海马齿状回神经前体细胞分裂及脑源性神经营养因子（BDNF）表达,Western蛋白印迹法检测海马BDNF蛋白表达水平。结果 免疫组化显示,慢性应激28 d小鼠海马齿状回神经前体细胞分裂减少,同时BDNF水平低下,均表现为阳性棕色颗粒缺失,豆腐果苷或盐酸氟西汀则明显增加神经前体细胞的分裂,并上调BDNF蛋白表达,提高BDNF水平。结论 豆腐果苷对慢性应激小鼠的抗抑郁作用机制可能与促进海马齿状回神经前体细胞增殖,提高BDNF水平有关。
     

藏药佐太在两种抑郁小鼠模型中的抗抑郁活性

目的通过行为绝望模型—悬尾实验(TST)、不可预测性慢性温和应激(CUMS)模型,探索藏药佐太(硫化汞、硫磺等煅制品的混合物)的抗抑郁作用。方法行为绝望模型—悬尾实验中,KM小鼠给予佐太14 d后,通过测定悬尾实验不动时间、开场实验(OFT)、血清中皮质酮(CORT)变化,评价佐太的抗抑郁活性。构建CUMS模型KM小鼠,给予佐太42 d,通过糖水偏好实验、开场实验和强迫游泳实验,进一步评价佐太的抗抑郁作用。通过ELISA法测定小鼠血清中5-羟色胺(5-HT)、去甲肾上腺素(NE)和海马中脑源性神经营养因子(BDNF)的水平。结果行为绝望模型—悬尾实验中,与空白组相比,佐太在剂量为60.697、303.49 mg/kg时能显著降低小鼠悬尾不动时间和血清中CORT水平。CUMS模型实验中,与CUMS模型组相比,佐太在剂量为6.069 7、60.697、606.97 mg/kg时,均可显著提高受试小鼠的糖水偏好率、开场实验中的自主活动,同时可显著降低强迫游泳实验(CFST)中的不动时间,表明佐太能缓解慢性应激引起的抑郁样行为。另外,佐太在上述3个剂量下,可显著增加CUMS模型小鼠血清中NE和海马中BDNF的水平,并在剂量为6.069 7、60.697 mg/kg下,可显著增加CUMS模型小鼠血清中5-HT的水平。结论藏药佐太在行为绝望模型和CUMS模型小鼠中表现出一定的抗抑郁活性,可能是通过降低CORT,提高5-HT、NE、BDNF的水平而发挥作用。     

基于雌激素受体通路探讨青娥丸对CUMS大鼠的抗抑郁机制

目的:研究青娥丸对慢性温和不可预知应激模型(CUMS)大鼠的抗抑郁效果,及其对雌激素受体及相关信号通路的调控作用,探讨青娥丸的抗抑郁机制。方法:54只SD大鼠建立CUMS抑郁大鼠模型,实验设为正常组、模型组、草酸依他普伦组(6.3 mg·kg^-1)及青娥丸低、中、高剂量组(1.71,5.13,15.39 g·kg^-1)。CUMS造模4周后给予各组相应药物治疗2周,采用行为学[糖水消耗实验(SPT),强迫游泳实验(FST),旷场实验(OFT)]评价大鼠抑郁状态;采用蛋白免疫印迹法(Western blot)检测雌激素受体α(ERα),雌激素受体β(ERβ),脑源性神经营养因子(BDNF)及酪氨酸激酶受体B(Trk B)的蛋白表达水平。结果:与正常组比较,模型组大鼠的糖水消耗率及旷场得分均下降(P<0.05,P<0.01),游泳不动时间显著延长(P<0.01),同时ERα,ERβ,BDNF和Trk B的蛋白表达水平下降(P<0.05,P<0.01);与模型组比较,各给药组大鼠的行为学表现得到改善,糖水消耗率及旷场得分增加(P<0.05,P<0.01),游泳不动时间减少(P<0.05),同时ERα,ERβ,BDNF和Trk B蛋白的表达明显上调(P<0.05,P<0.01),其中青娥丸中剂量组的调节作用更为显著。结论:青娥丸可改善CUMS模型大鼠的抑郁样行为,其机制可能与上调ERα,ERβ的表达,激活雌激素受体介导的ERβ/BDNF/Trk B通路起到神经保护作用有关。     

Antidepressant-like effects of Xiaochaihutang in a rat model of chronic unpredictable mild stress

Ethnopharmacological relevance

Xiaochaihutang (XCHT) has been used in China for thousands of years to treat “Shaoyang syndrome”, which involves depressive-like symptoms. However, few studies have investigated its antidepressant effects and pharmacological mechanism of action. The present study was designed to confirm the antidepressant effect of XCHT using a chronic unpredictable mild stress (CUMS) model and explore its potential mechanism of action by investigating the monoamine neurotransmitters (dopamine and 5-hydroxytryptamine) and neurotrophins (BDNF and NGF).

Materials and methods

The CUMS model was established in rats, and the antidepressant effect of XCHT (0.6, 1.7 and 5 mg/kg/day, given by gastric gavage for 4 weeks) was investigated using the open field test (OFT), food consumption test and sucrose preference test. The concentrations of 5-HT and DA in the hippocampus were measured by high performance liquid chromatography with electrochemical detection (HPLC-ECD). The expressions of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and their receptors tyrosine receptor kinase B (TrkB) and tyrosine receptor kinase A (TrkA) in the hippocampus were measured by immunohistochemical staining analysis.

Results

CUMS caused a significant decrease in OFT, food consumption and sucrose preference in rats, and these depression-like behaviors were significantly improved by XCHT (1.7 and 5 g/kg/day). Moreover, XCHT significantly increased the concentrations of 5-HT (0.6 and 5 g/kg/day) and DA (5 g/kg/day), and improved the BDNF, NGF, TrkB and TrkA expressions in the hippocampus (1.7 and 5 g/kg/day), which was reduced in CUMS rats.

Conclusion

The results obtained suggested that XCHT may have therapeutic actions on depression-like behavior induced by CUMS in rats possibly mediated by increasing the monoamine neurotransmitter concentration and neurotrophin expression in the hippocampus.     

人参皂苷对慢性应激抑郁模型大鼠行为学及HPA轴、BDNF的影响

目的:探讨人参皂苷对慢性应激所致大鼠抑郁模型的干预作用.方法:通过测定大鼠血清中皮质酮(COR)、糖皮质激素受体(GR)、盐皮质激素受体(MR)和脑组织中神经营养(BDNF)的mRNA表达水平,探讨人参皂苷的抗抑郁机制.结果:与正常组大鼠比较,经过慢性应激6周后大鼠糖水偏好显著下降,强迫游泳测试不动时间明显增加,表明慢性应激导致大鼠产生抑郁样行为.同时,抑郁大鼠的血清COR水平增加,海马GR、海马及皮层BDNF的mRNA表达水平均明显降低.给予人参皂苷(12.5,25,50 mg·kg-1)6周后,发现其能显著改善由慢性应激所致的大鼠抑郁行为及生化指标.各个组间海马MR的mRNA表达水平没有显著差异.结论:人参皂苷的抗抑郁作用机制可能通过调节下丘脑-垂体-肾上腺轴功能,进而提高脑组织BDNF 表达水平.     

人参拮抗高皮质激素血症致神经元损伤的作用

目的:探讨人参保护高皮质激素血症致神经元损伤的作用机制。方法:皮下注射皮质酮建立高皮质激素血症的动物模型,在模型的制备过程中同时ig给予高、中、低(7.2,3.6,1.8 g.kg-1)3个剂量的人参水煎剂,实验进行4周,行为学检测后处死动物,取脑剥离海马,对神经微丝蛋白(NF-L)、突触囊泡总蛋白(SYP)、脑源性神经营养因子(BDNF)、胶质源性神经营养因子(GDNF)进行蛋白免疫印迹。结果:模型组动物在行为学检测中,静止不动时间明显增加,与正常组有显著差别;体重较正常组显著降低并有显著差异。相关蛋白检测NF-L,SYP,BDNF,GDNF表达水平与正常组比较显著下调。人参高、中剂量组能够改善动物行为学变化及体重变化,人参各剂量组对神经元蛋白及神经营养因子均表现出明显的上调作用。结论:人参可通过上调神经营养因子的表达发挥神经元保护作用。     

Ethanol extracts from Hemerocallis citrina attenuate the decreases of brain-derived neurotrophic factor,TrkB levels in rat induced by corticosterone administration

Ethnopharmacological relevance

Hemerocallis citrina, a traditional herbal medicine, has been used for the improvement of behavioral and emotional status in Eastern-Asia countries.

Aim of the study

Our previous studies have demonstrated that the ethanol extracts of H. citrina flowers (HCE) reversed the behavioral alterations and monoamine neurotransmitter dysfunctions in stressed mice. However, the relation of its antidepressant-like action with neurotrophic molecular expressions remains unknown.

Materials and methods

To clarify this, we explored the effect of HCE (32.5, 65, 130 mg/kg, p.o.) on the behavior, brain-derived neurotrophic factor (BDNF) and its receptor (TrkB) in depression-like rats induced by exogenous administration of the stress hormone corticosterone (40 mg/kg, s.c.).

Results

It was observed that repeated administration of corticosterone induced an elevation on the serum corticosterone levels, which caused the abnormalities observed in the sucrose preference test and forced swimming test (FST). Administration of HCE (65 and 130 mg/kg) reversed the changes above and up-regulated the BDNF and TrkB receptor protein expressions in the brain region of frontal cortex and hippocampus.

Conclusion

These findings confirm that HCE produce an antidepressant-like effect in corticosterone-induced depression-like model of rats and this effect is at least partly mediated by BDNF-TrkB signaling in the frontal cortex and hippocampus.     

Involvement of mTOR‐related signaling in antidepressant effects of Sophoraflavanone G on chronically stressed mice

SophoraflavanoneG (SG), an important prenylated flavonoid isolated from Sophoraalopecuroides.L, is effective for many illnesses. The present study was designed to investigate whether the compound could reverse depressive‐like symptoms and investigate its possible mechanisms. Chronic Unpredictable Mild Stress (CUMS) mice were treated with fluoxetine and SG. The immobility time in forced swimming test (FST) and tail suspension test (TST) were recorded. The levels of pro‐inflammatory cytokines and neurotransmitters in the hippocampus were evaluated. Furthermore, the protein expressions of PI3K, AKT, mTOR, p70S6K, BDNF, and Trkb in hippocampus were detected. Rapamycin, the selective mTOR inhibitor, was used to estimate the potential mechanism. As a result, after 7 days of SG treatment, the immobility time in FST and TST was declined obviously. The levels of IL‐6, IL‐1β, and TNF‐α in the hippocampus were significantly reduced, and the quantity of 5‐HT and NE was raised considerably in SG‐treated group compared with the CUMS‐exposed group. Additionally, SG could up‐regulate the expressions of PI3K, AKT, mTOR, 70S6K, BDNF, and Trkb. The blockade of mammalian target of rapamycin signaling blunted the antidepressant effect and reversed the up‐regulation of BDNF expression caused by SG. These findings suggested that SG treatment alleviated depressive‐like symptoms via mTOR‐mediated BDNF/Trkb signaling.     

Ethanol extracts from Hemerocallis citrina attenuate the upregulation of proinflammatory cytokines and indoleamine 2,3-dioxygenase in rats

Ethnopharmacological relevance

Hemerocallis citrina, a traditional herbal medicine, has been used for the improvement of behavioral and emotional status in Eastern-Asia countries. Previous studies in our laboratory demonstrated that ethanol extracts from Hemerocallis citrina (HCE) enhanced monoamines and brain-derived neurotrophic factor (BDNF) in depression-like model of rodents.

Materials and methods

The present study extends earlier works on the role of anti-inflammation in regulating the antidepressant-like actions of HCE in rats exposed to chronic unpredictable mild stress (CUMS). Frontal cortex and hippocampal proinflammatory cytokines levels and indoleamine 2,3-dioxygenase (IDO) activity were measured after 4-week HCE treatment in the CUMS an control rats.

Results

Chronic administration of HCE reversed the decreased sucrose preference in sucrose preference test. In addition, we also found that HCE inhibited interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) expression, as well as IDO activity in frontal cortex and hippocampus, which were increased in rats exposed to CUMS.

Conclusions

Combining with our previous studies, our present finding suggests that the anti-inflammatory property of HCE might play a crucial role in its antidepressant-like effect through, at least in part, the restoration or improvement of monoaminergic and neurotrophin systems.