Protective effect of panaxatriol saponins extracted from Panax notoginseng against MPTP-induced neurotoxicity in vivo

Aim of the study

Panaxatriol saponins (PTS), the main constituents extracted from Panax notoginseng, a Chinese herbal medicine, has been shown to be an effective agent on various diseases. Our previous study has demonstrated that PTS is an inducer of thioredoxin-1 (Trx-1) and has a possible potential as a therapeutic agent for Parkinson's disease (PD). However, the effect of PTS on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity in vivo is unknown.

Materials and methods

Using locomotor activity test and traction test, we detected the effect of PTS on MPTP-induced behavioral impairment. Tyrosine hydroxylase, Trx-1, cyclooxygenase-2, pro-caspase-9, pro-caspase-12 and caspase-3 expressions in the anatomical region of substantia nigra pars compacta (SNc) were tested by Western blot.

Results

PTS provided neuroprotection against the loss of dopaminergic neurons and behavioral impairment caused by MPTP. MPTP-induced neuronal death in the SNc was suppressed by PTS through increasing Trx-1 expression, suppressing cyclooxygenase-2 over-expression and inhibiting mitochondria-mediated apoptosis.

Conclusions

PTS, an inducer of Trx-1, has pluripharmacological properties in the protection against PD including enhancing antioxidant activity, acting as neurotrophic factor, modulating inflammation and inhibiting mitochondria-mediated apoptosis.     

Calycosin attenuates MPTP‐induced Parkinson's disease by suppressing the activation of TLR/NF‐κB and MAPK pathways

Parkinson is the second common neurodegenerative disease. The characteristics of Parkinson's disease (PD) are the dopamin neurons loss caused by neuroinflammation responses. C alycosin, an isoflavone phytoestrogen isolated from Astragalus membranaceus, has multiple pharmacological activities, such as anti‐inflammation, anti‐tumor, and neuroprotective effects. However, it is unknown whether calycosin can mitigate PD symptoms. This study aims to explore whether calycosin can alleviate PD symptoms and the underlying mechanisms. PD was induced in mice by 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) injection, and calycosin was given intracerebroventricularly to these mice. A cell model of nerve inflammation was established by BV2 microglia cells injected with lipopolysaccharide (LPS). The motor states were evaluated by stepping, whisker, and cylinder experiments. The states of dopaminergic neurons and microglia were detected by immunostainning of tyrosine hydroxylase and cluster of differentiation molecule 11b (CD11b). The expression levels of inflammatory factors were detected by qPCR. Toll‐like receptor (TLR)/nuclear factor kappa B (NF‐κB) and mitogen‐activated protein kinase (MAPK) pathways were investigated by western blot. We found that calycosin treatment mitigated the behavioral dysfunctions and inflammatory responses in MPTP‐induced PD mice. The TLR/NF‐κB and MAPK pathways in MPTP‐induced PD mice were inhibited by calycosin treatment, which was coincident with experiments in LPS‐induced BV2 cells. Above all, calycosin mitigates PD symptoms through TLR/NF‐κB and MAPK pathways in mice and cell lines.     

川芎茶调散对帕金森病小鼠多巴胺神经元损伤的保护作用及机制研究

目的:研究川芎茶调散对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)致小鼠多巴胺(DA)神经元损伤的保护作用,并从抗氧化角度探讨其神经保护作用的可能机制.方法:将3～5月龄的雄性C57/BL小鼠随机分为6组:正常组、川芎茶调散高、中、低(75,50,25 g·kg~(-1)生药量)3个剂量组、模型组.每组8只动物.观察爬杆实验测试行为学改变,HPLC检测小鼠纹状体中DA及其代谢产物含量,免疫组化检测小鼠黑质中TH阳性细胞数,荧光比色检测黑质中超氧化物歧化酶(SOD)活力.结果:高、中、低剂量川芎茶调散对帕金森氏病(PD)小鼠的行为损害均有一定的保护作用(P<0.01);川芎茶凋散高剂量组小鼠纹状体中DA含量及TH阳性神经元数量明显高于模型组(均P<0.05);川芎茶调散中、高剂量组相对于模型组,SOD活力显著增高(P<0.05).结论:川芎茶调散可明显改善MPTP所致的小鼠PD模型的运动障碍,同时能对MPTP引起的DA神经元损伤起到保护作用,川芎茶调散对DA神经元的保护作用可能与其较强的抗氧化能力有关.     

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??OBJECTIVE To investigate the effect of minocycline on the behavior damage induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP) in C57BL/6 mice. METHODS The PD models were formed with intraperitoneal injections of MPTP in C57BL/6 mice.Forty mice were randomly divided into 4 groupscontrol group,MPTP group,MPTP+Mino group and Mino group(10 for each group). The levels of midbrain DA,DOPAC and HVA were measured by HPLC after behavior tests(swimming test,pole test and locomotor activity test ) were performed. RESULTS Swimming scores,climbing score and locomotor activity of MPTP+Mino group were significantly higher than the MPTP group(P<0.01). And the content of midbrain DA,DOPAC and HVA of MPTP+Mino group was significantly higher than the MPTP group(P<0.01). CONCLUSION Minocycline inhibits MPTP-induced damage of motor function in mice,the mechanism may be work via inhibiting MPTP-induced mice dopamine neuron damage.     

Acanthopanax senticosus Harms as a prophylactic for MPTP-induced Parkinson's disease in rats

The aim of this study was to determine whether Acanthopanax senticosus Harms (ASH) offers protection against Parkinson's disease (PD) and its related depressive behaviors in rats administered 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). We examined how ASH affected the MPTP-induced loss of tyrosine hydroxylase (TH)-positive neurons in the midbrain of rats. Extract from the stem bark of ASH prepared with hot water was dissolved in distilled water. Rats were then orally administered ASH (250 mg/kg) once a day for 2 weeks before ASH administration plus an intraperitoneal injection of MPTP (20 mg/kg). The pole test and catalepsy test were used to evaluate the effects of ASH administration on bradykinesia and depressive behaviors in the PD model of rats given MPTP for 2 weeks. Treatment with ASH for 2 weeks resulted in prophylactic effects on MPTP-induced Parkinsonian bradykinesia and catalepsy. Immunohistochemistical analysis using TH antibody showed that ASH provided cytoprotective effects against MPTP-induced loss of dopamine (DA) cells. The present results suggest that it may be possible to use ASH for the prevention of nigral degenerative disorders, e.g., PD with depression, caused by exposure to toxic substances.     

原儿茶酸对帕金森病模型小鼠中脑和纹状体D2DR、iNOS和TH表达的影响

目的考察原儿茶酸对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)致帕金森病(PD)小鼠中脑和纹状体多巴胺D2受体(D2DR)、诱导型一氧化氮合酶(i NOS)和酪氨酸羟化酶(TH)表达的影响。方法选用C57BL褐鼠为实验动物,连续7 d ip MPTP(25 mg/kg)制备PD动物模型。造模前后分别ip给予原儿茶酸(10 mg/kg)或阳性对照药美多芭(125 mg/kg)进行预防和治疗。应用Western blotting的方法,观察原儿茶酸对PD模型小鼠中脑和纹状体内D2DR、i NOS和TH表达的影响。结果 PD模型小鼠脑组织内i NOS表达升高,D2DR和TH表达下降,而原儿茶酸能够显著降低PD模型小鼠脑中i NOS的表达,提高D2DR和TH的表达量。结论原儿茶酸对MPTP诱导的PD模型小鼠有保护作用,可能通过提高PD模型小鼠中脑和纹状体内D2DR和TH的表达,降低i NOS的表达发挥作用。     

Effect of Zishenpingchan granule prepared from Chinese medicinal substances on the c-Jun N-terminal protein kinase pathway in mice with Parkinson's disease induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

OBJECTIVE:To investigate the regulatory mechanism of the c-Jun N-terminal protein kinase(JNK)signaling pathway in substantia nigra(SN) dopaminergic neurons inflammation and apoptosis, and the neuroprotective effect of Zishenpingchan granules in mice with Parkinson's disease(PD) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).METHODS:PD model mice were established by intraperitoneally injecting MPTP.Sixty mice were divided into a model group, Traditional Chinese Medicine(TCM) group and control group.The mice of the TCM group were administered Zishenpingchan granules 7 days before PD induction.Seven days after PD induction, we examined locomotor activity,and performed the rotarod test and swimming test,to evaluate limb movement function.Furthermore,we used immunohistochemistry and western blotting to examine the expression of tyrosine hydroxylase(TH), cyclooxygenase-2(Cox-2), caspase-3 and p-JNK.The terminal deoxynucleotidyl transferase mediated d UTP nick end labeling(TUNEL) method was used to examine neuron apoptosis in the SN.RESULTS:Compared with the control group, the mean score of locomotor activity, rotarod test and swimming test was significantly lower in the model group(P 0.05); the TH-positive neuron expression was significantly decreased in the SN pars compacta(SNpc); the protein expression levels of Cox-2,caspase-3 and p-JNK was obviously increased; and the number of TUNEL-positive neurons in the SN was increased(P 0.01).Compared with the model group, the mean score of neurobehavioral tests in the TCM group was obviously higher, the loss of TH-positive neurons ignificantly decreased, the protein expression levels of Cox-2, caspase-3 and p-JNK obviously decreased, and the number of TUNEL-positive neurons in the SN clearly decreased(P 0.01).CONCLUSION:The JNK pathway plays an important role in the regulation of inflammation and apoptosis in nigral cells in PD mice.TCM can suppress the over-activation of the JNK pathway in the SN, and alleviate the inflammatory response in nigral cells and dopaminergic neuron apoptosis in PD mice.     

大补阴丸 牵正散及合方对MPTP诱导帕金森小鼠的神经保护作用

目的:探讨大补阴丸、牵正散及合方对帕金森病的神经保护的作用。方法:采用腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)建立帕金森病模型。C57BL/6雄性小鼠随机分为正常组、模型组、大补阴丸组、牵正散组和合方组。采用爬杆法测量小鼠行为学改变;于造模14天应用高压液相电化学内标法测定前脑单胺类神经递质含量;用免疫组织化学技术检测黑质酪氨酸羟化酶阳性神经元数目;透射电镜观察黑质神经元超微结构的变化。结果:爬杆实验显示大补阴丸、牵正散及合方能明显改善PD小鼠的行为学(P0.05),减少黒质多巴胺能神经元丢失(P0.01),减轻黑质神经元核膜、线粒体等结构的损伤。但是给药14天后,中药组小鼠相对PD小鼠前脑内的单胺类神经递质水平并没有明显改变。结论:大补阴丸,牵正散及合方对MPTP诱导PD小鼠黒质多巴胺能神经元有一定保护作用。     

Combined prescription (OAH19T) of Aralia cordata Thunb and Cimicifuga heracleifolia Komar and its major compounds inhibit matrix proteinases and vascular endothelial growth factor through the regulation of mitogen-activated protein kinase pathway

Aim of the study

Polygalae radix, the root of Polygala tenuifolia Willd, has commonly been used for the treatment of amnesia and anxiety in traditional Korean medicine. The aim of this study was to investigate its neuroprotective effects and possible mechanisms of action in models of Parkinson's disease.

Materials and methods

This study utilized a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, a reactive oxygen species (ROS) assay, a nitric oxide (NO) production assay, and a caspase-3 activity test as measures of cell viability in PC12 cells damaged by 6-hydroxydopamine (6-OHDA). The protective effects of PRE against 1-methyl-4-phenylpyridium (MPP⁺)-induced neurotoxicity were assessed in rat primary dopaminergic neurons and in a mouse PD model in which PRE was administered (100 mg/kg/day, 3 days, p.o.) before acute 1-mehtyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) toxicity. Finally, TH immunohistochemistry tests were conducted in the substantia nigra pars compacta (SNpc) and striatum (ST).

Results and conclusions

PRE significantly inhibited 6-OHDA-induced cell damage at doses of 0.05-1 μg/ml with a maximal effect at 0.1 μg/ml. Caspase-3 activity and the production of ROS and NO were alleviated at 0.1 μg/ml. Also at this dose, PRE protected mesencephalic dopaminergic neurons from MPP⁺-induced toxicity. In an in vivo mouse model of PD, PRE protected dopaminergic neurons and fibers from MPTP-induced toxicity in the SNpc and ST. These results demonstrate that PRE has protective effects on dopaminergic neurons via its anti-oxidant and anti-apoptotic activity.     

苁蓉总苷对MPTP帕金森病模型小鼠脑黑质多巴胺能神经元保护作用的研究

目的　探讨苁蓉总苷对MPTP帕金森病(PD)模型小鼠脑黑质多巴胺能(DA)神经元的保护作用。方法 将小鼠随机分为正常组、模型组、苁蓉总苷高（400 mg/kg）、中（200 mg/kg）、低（100 mg/kg）剂量组,各组在造模前3天灌服相应药物;采用MPTP（30 mg/kg）连续5天腹腔注射制作小鼠慢性PD模型,运用爬竿试验定量组织化学检测小鼠的神经行为,免疫组化法测定小鼠脑纹状体酪氨酸羟化酶（tyrosine hydroxylase,TH）阳性纤维表达以及黑质TH神经元数量的变化。结果 在造模后7天和14天时,与模型组比较,苁蓉总苷高剂量组爬杆时间缩短（P＜0.01）,苁蓉总苷各剂量组纹状体TH阳性纤维的平均光密度值均明显增高（P＜0.01）;3个剂量组的苁蓉总苷均能对抗MPTP导致的TH阳性神经元数量减少,但只有高剂量组在两个时间点上与模型组的差异有统计学意义（P＜0.05）。结论 苁蓉总苷能够显著改善PD模型小鼠的神经行为、抑制脑黑质DA神经元的数量减少和纹状体TH表达的降低。     

电针对1-甲基-4-苯基-1,2,3,6-四氢吡啶诱导的帕金森病模型小鼠线粒体功能的影响

目的:观察电针对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病(PD)模型小鼠行为学、酪氨酸羟化酶(T H)、线粒体复合物Ⅰ — Ⅳ活性、线粒体膜电位、线粒体超微结构的影响,探讨电针治疗PD的可能靶点及机制.方法:将雄性C57BL/6小鼠随机分为对照组、模型组、美多巴组及电针组,每组11只,采用...     

蝎毒耐热蛋白对帕金森病模型小鼠脑内一氧化氮合酶的影响

目的观察蝎毒耐热蛋白(SVHRP)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)致帕金森病小鼠运动协调性和脑内神经元型一氧化氮合酶(nNOS)免疫反应(IR)阳性神经元的影响。方法C57BL/6小鼠颈部scMPTP20mg/kg,连续8d造模型。同时设立SVHRP治疗组,测定小鼠爬竿和游泳时间以检测其运动协调性。应用免疫细胞化学方法观察脑内黑质酪氨酸羟化酶(TH)和nNOS免疫反应阳性神经元的变化。结果MPTP致模型小鼠在黑质致密部多巴胺能神经元损伤后,运动协调能力下降,同时在尾核的nNOS免疫反应阳性神经元细胞数较正常组增多。治疗组小鼠运动协调能力与正常对照组比较没有明显改变,尾核的nNOS免疫反应阳性神经元细胞数较模型组明显减少。结论SVHRP可以保护中脑黑质致密部多巴胺能神经元及改善MPTP引起的运动协调性降低,逆转MPTP引起的尾核nNOS免疫反应活性增强。而NO可能参与其保护机制。     

咖啡酸苯乙酯对神经毒素诱导帕金森病模型神经元损伤的保护作用

目的:探讨咖啡酸苯乙酯(Caffeic Acid Phenethyl Ester,CAPE)对神经毒素诱导的帕金森病模型神经元损伤的保护作用。方法:体外用MPP+处理PC12细胞制作PD细胞模型,利用MTT法和流式细胞仪分别测定细胞存活率和细胞凋亡。将C57BL雄性小鼠用MPTP处理(20 mg/kg,3次,ip)制作PD小鼠模型,用尼氏染色检测黑质致密区神经细胞数,免疫组化方法检测黑质致密区酪氨酸羟化酶(TH)和Caspase-3阳性细胞数。结果:PC12细胞中,1.25μM的CAPE可以显著抑制MPP+诱导的细胞活性的丧失和细胞凋亡。PD小鼠模型中,MPTP模型组黑质致密区神经细胞数和TH阳性细胞数较正常对照组明显降低,黑质致密区Caspase-3阳性细胞数较正常对照组明显增加。CAPE(50 mg/kg,ip,7d)可以显著增加黑质致密区TH阳性细胞数,减少黑质致密区Caspase-3阳性细胞数。结论:CAPE对神经毒素诱导的PD模型神经元损伤有明显的保护作用。     

针刺筋会穴阳陵泉对帕金森模型小鼠黑质TH和GFAP表达的影响

目的观察比较1-甲基-4-苯基-1,2,3,6四氢吡啶（MPTP）诱导的帕金森小鼠模型及其接受针刺治疗后黑质酪氨酸羟化酶（TH）和星形胶质细胞的胶质原纤维性蛋白（GFAP）的表达变化。方法以腹腔注射（30 mg/kg）MPTP诱导7 d形成帕金森小鼠模型,针刺双侧筋会穴＂阳陵泉＂及＂舞蹈震颤区＂,每日1次共治疗21 d。针刺结束后,采用免疫荧光组化检测小鼠脑黑质TH的和GFAP的表达变化。结果在7 d造模后,模型组和针刺组TH阳性细胞显著丢失;治疗结束后,与模型组比较,针刺组TH阳性细胞数量增加,正常组和针刺组GFAP的阳性细胞数量减少。结论 MPTP可促进帕金森模型小鼠表达;针刺筋会穴阳陵泉可对MPTP诱导小鼠黑质多巴胺能神经有保护作用,能明显地减弱MPTP伤害性刺激引起的行为反应。     

针刺对帕金森病小鼠行为学及纹状体神经元凋亡的影响

目的:观察针刺对帕金森病模型小鼠行为学和纹状体神经元凋亡的影响,探讨针刺治疗帕金森病的机制。方法:成年C57/BL雄性小鼠28只,随机分为正常组、模型组、针刺组和西药组。腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶复制帕金森病模型。针刺组给予"百会"风府"针刺及"阳陵泉"电针治疗,每日1次,每次15min,治疗14d;西药组给予多巴丝肼灌胃治疗,每日1次,治疗14d。分别在造模前、造模后和治疗后对各组小鼠进行自主活动计数、爬杆实验、游泳实验的行为学检测,采用TUNEL法测定各组小鼠纹状体神经元凋亡指数。结果:模型组小鼠造模后自主活动计数、游泳实验评分显著低于正常组(P<0.01),爬杆实验评分显著高于正常组(P<0.01);针刺组、西药组小鼠治疗后自主活动计数、游泳实验评分显著高于模型组(P<0.05,P<0.01),爬杆实验评分显著低于模型组(P<0.05)。模型组、针刺组、西药组纹状体神经元凋亡指数均显著高于正常组(P<0.01),针刺组凋亡指数显著低于模型组(P<0.01)。结论:针刺"百会"风府"阳陵泉"可改善小鼠自主活动、爬杆、游泳等行为,减少纹状体神经元凋亡,改善帕金森病的运动症状。     

THC (Δ9‐Tetrahydrocannabinol) Exerts Neuroprotective Effect in Glutamate‐affected Murine Primary Mesencephalic Cultures Through Restoring Mitochondrial Membrane Potential and Anti‐apoptosis Involving CB1 Receptor‐dependent Mechanism

Aging‐related neurodegenerative diseases, such as Parkinson's disease (PD) or related disorders, are an increasing societal and economic burden worldwide. Δ9‐Tetrahydrocannabinol (THC) is discussed as a neuroprotective agent in several in vitro and in vivo models of brain injury. However, the mechanisms by which THC exhibits neuroprotective properties are not completely understood. In the present study, we investigated neuroprotective mechanisms of THC in glutamate‐induced neurotoxicity in primary murine mesencephalic cultures, as a culture model for PD. Glutamate was administered for 48 h with or without concomitant THC treatment. Immunocytochemistry staining and resazurin assay were used to evaluate cell viability. Furthermore, superoxide levels, caspase‐3 activity, and mitochondrial membrane potential were determined to explore the mode of action of this compound. THC protected dopaminergic neurons and other cell types of primary dissociated cultures from glutamate‐induced neurotoxicity. Moreover, THC significantly counteracted the glutamate‐induced mitochondrial membrane depolarization and apoptosis. SR141716A, a CB₁ receptor antagonist, concentration‐dependently blocked the protective effect of THC in primary mesencephalic cultures. In conclusion, THC exerts anti‐apoptotic and restores mitochondrial membrane potential via a mechanism dependent on CB₁ receptor. It strengthens the fact that THC has a benefit on degenerative cellular processes occurring, among others, in PD and other neurodegenerative diseases by slowing down the progression of neuronal cell death. Copyright © 2016 John Wiley & Sons, Ltd.     

Neuroprotective effects of kukoamine A on 6-OHDA-induced Parkinson’s model through apoptosis and iron accumulation inhibition

Objective: Parkinson’s disease(PD) is characterized by the loss of dopaminergic neurons in substantia nigra(SN). Our previous study demonstrated kukoamine A(KuA) to exhibit strong neuroprotective effects through antioxidative stress, and autophagy in MPTP/MPP+-induced PD models in vivo and in vitro. It is necessary to evaluate the efficacy of the anti-PD effects under various models.Methods: In the present study, total chemical synthesis was used to obtain KuA, which performed low content in Lycii Cortex. Then, 6-OHDA-induced PD model of PC12 cells was used to investigate the effects of KuA on PD.Results: Our results demonstrated that KuA ameliorated cell loss and mitochondrial membrane potential(MMP) loss, and inhibited Bax/Bcl-2 ratio increase that were induced by 6-OHDA. Iron accumulation in SN is thought to participate in neuronal death in PD, which subsequently resulted in oxidative stress and overexpression of a-synuclein caused by iron metabolism protein disorder. In our study, KuA could chelate cellular iron content and decrease iron influx. Moreover, KuA could upregulate the expression of ferroportin1 and Hephaestin, downregulate the expression of DMT1, TfR, and Ferritin to maintain cellular iron homeostasis avoiding neuronal death from cellular iron deposition. Moreover, KuA could decrease the expression of a-synuclein in cells. All the results indicated that KuA protected against neurotoxininduced PD due to the apoptosis inhibition and iron homeostasis maintaining.Conclusion: KuA treatment might represent a neuroprotective treatment for PD.     

黄芩苷对MPTP帕金森病小鼠的保护作用

目的观察黄芩苷对1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）帕金森病小鼠的保护作用。方法以10月龄（老龄）C5,BL小鼠腹腔注射MPTP（每天30mg／kg）3天制备小鼠帕金森病模型;黄芩苷组灌胃黄芩苷（每天100mg／kg,共15天）;用悬挂、游泳实验检测小鼠肢体运动功能;HPLC法测定小鼠脑纹状体多巴胺（DA）含量;分光光度法测定小鼠脑组织谷胱苷肽（GSH）、谷胱苷肽过氧化酶（GSH-Px）及丙二醛（MDA）的含量;小鼠脑冰冻切片,行酪氨酸羟化酶（TH）免疫组化染色,观察中脑黑质多巴胺能神经损伤情况。结果（1）模型组小鼠悬挂和游泳实验分值及脑纹状体DA含量均显著降低,黑质TH阳性细胞显著丢失,说明模型复制成功。（2）黄芩苷能够防止帕金森病小鼠黑质多巴胺能神经细胞丢失,防止模型小鼠纹状体DA含量的降低,并显著升高小鼠脑内GSH水平。但短时间给黄芩苷不能明显改善MPTP致帕金森病小鼠运动功能障碍。结论黄芩苷预防给药对MPTP致帕金森病小鼠有保护作用。     

Dangguijakyak-san protects dopamine neurons against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity under postmenopausal conditions

Dangguijakyak-san protects dopamine neurons against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity under postmenopausal conditions.

Ethnopharmacological relevance

Dangguijakyak-san (DJS), a famous traditional herbal formula, has long been used to treat gynecological disorders, including postmenopausal symptoms. This study evaluated the effects and mechanism of DJS on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity in a postmenopausal mouse model induced by ovariectomy.

Materials and methods

Three weeks after ovariectomy, C57bl/6 female mice were divided randomly into (1) control, (2) MPTP (30 mg/kg/day, i.p., 5 days), (3) MPTP + estrogen (50 μg/kg/day, i.p., 5 days), and (4) MPTP + DJS (50 mg/kg/day, p.o., 5 days) groups. We investigated the behavioral recovery and dopamine neuron protection of DJS using the pole test and tyrosine hydroxylase (TH) immunohistochemistry. We also explored the mechanism by assessing the protein expression of Bax, Bcl-2, cytochrome c, and cleaved caspase-3.

Results

DJS treatment restored the movement behavior impaired by MPTP, showing a similar or better effect than estrogen. DJS protected TH-immunoreactive cells and fibers in the nigrostriatal region from MPTP toxicity. In addition, DJS inhibited the Bcl-2 decrease and Bax increase in mitochondria, cytochrome c release to the cytosol, and caspase-3 activation induced by MPTP.

Conclusions

DJS showed behavior recovery and dopamine neuron protection against MPTP-induced toxicity via anti-apoptotic activities in ovariectomized female mice. These results suggest that DJS treatment is effective for postmenopausal neurodegenerative diseases.     

川芎嗪对MPTP诱导的多巴胺能神经元毒性的影响

目的 观察川芎嗪体外、体内对MPTP)导致的多巴胺能(dopaminergic,DA)神经元毒性的影响.方法 体外实验,提取孕14 d的Sprague-dawley大鼠胎鼠黑质区多巴胺神经元,MPP<'+>诱导其损伤,运用免疫荧光法测定黑质酪氨酸羟化酶(tyrosine hydroxylase,TH)阳性神经元数量及状态变化;体内实验,以C57BL/6J小鼠为研究对象,采用腹腔注射MPTP方法制作小鼠慢性帕金森(Parkinson's disease,PD)模型,运用免疫组化法测定TH阳性神经元数量变化.结果 体外实验:TMP组多巴胺神经元形态数目均有所改善,其中500μmol/L组与模型组相比具显著性差异.体内实验:与模型组相比,注射TMP后,多巴胺神经元密度有所增加.结论 TMP能不同程度地改善MPTP诱导的多巴胺神经元的损伤.