MPTP损伤的小鼠PD模型的制作与评价

帕金森病(Parkinson!sdisease,PD)动物模型研究的目的是揭示多巴胺能神经元特异性损伤的机制,进而探索针对这种损伤的神经保护方法或治疗方法.由神经毒素MPTP损伤的小鼠PD模型,广泛应用于散发性PD的研究中.根据注射总剂量、两次注射间隔时间、注射方式的不同,制成了适合于不同研究目的的各种小鼠PD模型.关于MPTP导致的PD模型动物神经损伤的评价方式也是多层面、多指标并存的.对MPTP动物模型的起源和MPTP导致多巴胺能神经元损伤途径进行了较为系统的概述,并对MPTP小鼠PD模型的制作方法与评价指标进行较为详细的归纳.     

慢性间断性缺氧对帕金森病模型小鼠行为学及纹状体多巴胺含量的影响

目的:探讨慢性间断性缺氧(chronic episodic hypoxia,EHYP)对帕金森病(Parkinson disease,PD)模型小鼠行为学及纹状体多巴胺(DA)含量的影响。方法:44只雄性6周龄C57BL/6小鼠随机分为百草枯 EHYP组、EHYP组、百草枯组和对照组,观察小鼠自发行为活动及悬挂实验、游泳实验、步态实验进行行为学检测,高效液相色谱分析(HPLC)测定纹状体DA含量。结果:百草枯 EHYP组小鼠出现PD综合征表现,悬挂实验及游泳实验结果与其它各组间有显著性差异(P<0.05或P<0.01),纹状体DA含量较其它各组小鼠出现明显下降(均P<0.01),余各组行为学实验结果及纹状体DA含量差异无显著性(P>0.05)。结论:EHYP联合百草枯暴露可使小鼠出现PD综合征表现及纹状体DA含量的明显下降。     

纹状体胆碱能神经元功能失调与帕金森病

纹状体作为基底核中最大的核团,在基底神经节生理和病理的调控中起到重要作用,尤其在帕金森病(PD)的病理中扮演着重要的角色,其中胆碱能中间神经元(CINs)是纹状体中重要的调控神经元。尽管以往人们已经认识到纹状体多巴胺/乙酰胆碱失衡是PD的关键神经化学基础,由于对CINs的解剖和功能特征认识的局限性和缺乏特异性胆碱能受体亚型药物或工具,制约着针对CINs的PD治疗。然而,随着研究的不断进行,人们对于纹状体CINs功能障碍对PD潜在的影响越来越关注。本文综述了CINs一些关键的形态、电生理学特性、突触和受体功能特点及其在PD状态下的变化,为如何更好地进行PD治疗提供了新途径。     

腺苷受体拮抗剂对帕金森病小鼠神经递质的影响

运用神经毒剂ＭＰＴＰ（１－ｍｅｔｈｙ－４－ｐｈｅｎｙ－１,２,３,６－ｔｅｔｒａｈｙｄｒｏｐｙｒｉｄｉｎｅ）制作的ＩＣＲ小鼠帕金森病模型,通过荧光测定方法和免疫组织化学方法,测定ＭＰＴＰ及腺甘Ａ２ａ受体拮抗剂喹唑啉（Ｑｕｉｎａｚｏｌｉｎｅ,ＣＰ６６７１３）对单胺类神经递质去甲肾上腺素（ＮＡ）、多巴胺（ＤＡ）、５－羟色胺（５－ＨＴ）及其代谢产物５－羟吲哚乙酸（５－ＨＩＡＡ）和氨基酸类神经递质γ－氨基     

高频刺激丘脑底核对帕金森病大鼠模型纹状体NOS阳性神经元的影响

目的观察高频刺激丘脑底核对帕金森病(PD)大鼠纹状体中NOS阳性神经元的影响,以探求其作用机制。方法应用6OHDA制备偏侧PD大鼠模型,丘脑底核区埋入刺激电极进行电刺激,采用组织化学方法观察纹状体中NOS阳性神经元的变化。结果PD大鼠纹状体中NOS阳性神经元数与正常大鼠相比明显增加(P<0.01),经电刺激后PD大鼠纹状体NOS阳性神经元数量明显减少,且与正常大鼠相比无显著性差异。结论高频电刺激丘脑底核治疗PD的机制之一可能是与其抑制纹状体NO的过度释放有关。     

环加氧酶-2对帕金森病小鼠黑质多巴胺能神经元的影响

目的和方法 :选用C5 7BL种系环加氧酶 2 (cyclooxygenase 2 ,COX 2 )缺陷小鼠 ,腹腔注射 1 甲基 4 苯基 1,2 ,3,6 四氢吡啶 (MPTP)制备帕金森病小鼠模型 ,用免疫组织化学方法观察COX 2对帕金森病小鼠黑质多巴胺能神经元的影响。结果 :行为学及免疫组织化学观察显示 ,野生型帕金森病小鼠的死亡率明显高于COX 2缺陷杂合子帕金森病小鼠 (P <0 .0 1) ,野生型帕金森病小鼠黑质致密部酪氨酸羟化酶 (tyrosinehydroxylase,TH)免疫反应阳性神经元数目较杂合子帕金森病小鼠明显减少 (P <0 .0 1)。结论 :COX 2可能与帕金森病时黑质多巴胺能神经元的损伤有关     

小鼠晶体蛋白质组的双向电泳和质谱鉴定研究

目的应用双向电泳和质谱技术研究5周龄小鼠晶体蛋白质组。方法提取小鼠晶体总蛋白,进行固相pH梯度（IPG）等电聚焦双向电泳,胶体考马斯亮蓝R-250染色,使用PDQuest7．30图像分析软件分析电泳图像。选择主要蛋白点胶上酶解,应用基质辅助激光解析电离飞行时间／飞行时间（MALDI—TOF／TOF）仪器进行串联质谱（MS／MS）鉴定。结果上样量为882μg和190μg时,分别检测370±41蛋白点（n=3）和57±5个蛋白点（n=3）。高上样量能够较好地分离晶体低丰度蛋白,如念珠状纤维结构蛋白BFSP;低上样量可很好地分离高丰度蛋白-晶体蛋白（包括αA、αB;βA1～βA4;βB1～βB3;γA～γF和γS等）。质谱鉴定得到1种细胞骨架蛋白和16种高丰度晶体蛋白。结论双向电泳和质谱技术有效考察了晶体总蛋白质,为分析白内障形成过程中蛋白质的表达改变提供了新的方法和途径。     

高频刺激丘脑底核对帕金森病大鼠模型纹状体神经元放电的影响

目的:观察高频刺激丘脑底核(STN)对帕金森病(PD)大鼠模型纹状体 (STR)神经元自发放电的影响.方法:应用6-羟基多巴胺(6-OHDA)制备偏侧PD大鼠模型,丘脑底核区插入刺激电极进行高频刺激,采用细胞外单位记录的方法观察STR神经元自发放电频率的改变.结果:正常大鼠刺激后STR神经元反应主要以兴奋型反应为主, PD大鼠STR神经元反应主要以兴奋抑制型为主,且随着刺激时间的延长,抑制持续时间逐渐增加,持续时间与刺激时间密切相关(r=0.94).结论:刺激STN可使PD大鼠纹状体的异常放电得到改善,提示高频电刺激STN可作为一种有效的治疗PD的方法.     

胚鼠黑质细胞分别移植于帕金森病模型鼠纹状体和侧脑室的比较研究

胚鼠黑质细胞悬液分别移植于帕金森病（ＰＤ）鼠纹状体和侧脑室。移植后两组动物的Ａｐｏｍｏｒｐｈｉｎｅ诱导旋转行为均得到极明显改善,移植细胞生长发育良好。移植细胞和宿主细胞间的信息联系,在纹状体内可能以突触传递方式为主。侧脑室内移植的黑质细胞,相当于人工放置的“接触脑脊液神经元”,可能主要通过非实触传递方式而发挥作用。     

中脑多巴胺A9、A10神经元在MPTP损伤中敏感性的差异

自从科学家发现1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）选择性损伤中脑黑质致密区（A9）多巴胺神经元通路以来,用这种神经毒素诱导的帕金森病（PD）模型被广泛应用于PD的研究。然而同为DA能神经元的中脑一边缘一皮层的DA（A10）系统是否也遭到同样损伤呢？     

MPTP帕金森病动物模型研究进展

用神经毒素1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)制备的动物模型,无论在神经生化和病理组织学特征,还是在运动行为表现方面都酷似人帕金森病(PD),是目前研究PD的理想模型。对MPTP动物模型发病机制等方面的深入研究将有助于PD的防治。     

MPTP Decreases MT-I mRNA In Mouse Striatum

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a drug that induces parkinsonism in humans and non-human primates. Free radicals are thought to be involved in its mechanism of action. Recently, metallothionein has been proposed to play a role as a scavenger of free radicals. In the present work, we studied the effect of MPTP neurotoxicity on brain metallothionein-I (MT-I) mRNA expression. Male C-57 black mice were treated with MPTP (30 mg/kg, i.p., daily) for 3 or 5 days. All animals were killed by cervical dislocation 7 days after the last MPTP dose. The brains were removed quickly and immediately frozen, and quantitative in situ hybridization was performed using MT-I cDNA probe. MT-I mRNA content in striatum, a region which is known to be highly predisposed and sensitive to MPTP-induced oxidative stress, decreased by 30% (3 days) and 39% (5 days) respectively, after the last MPTP administration. These results suggest that MT-I gene expression is decreased in MPTP neurotoxicity. It is suggested that the reduction of MT, an anti-oxidant and a free radical scavenger, in the striatum by MPTP enables the neurotoxin to exert maximal oxidative damage to the striatum.     

MPTP as a Mitochondrial Neurotoxic Model of Parkinson's Disease

1-Methy-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a potent neurotoxin extensively used to model Parkinson's disease (PD). A cascade of deleterous events, in which mitochondria play a pivotal role, drives MPTP neurotoxicity. How mitochondria are affected by MPTP and how their defect contributes to the demise of dopaminergic neurons in this model of PD are discussed in this review.     

游泳运动对神经毒素引起的小鼠运动机能损伤的保护作用

目的观察游泳运动对神经毒素(MPTP)致小鼠神经和运动机能损伤的保护作用,探讨可能存在的机制。方法在注射MPTP或生理盐水前1d和注射后1、4、7、10d,测量MPTP游泳组、MPTP非游泳组和生理盐水对照组小鼠的爬杆时间和步伐,第11天用放射自显影法测定纹状体多巴胺转运体密度。结果MPTP两组第1天步伐延长,随后恢复。第4天MPTP非游泳组步伐小于生理盐水组和游泳组,后两组差异无显著性。MPTP两组第1天爬杆时间延长,但与生理盐水组比较差异无显著性。游泳组在随后各时间点爬杆时间依次缩短,并在第7、10天明显短于MPTP非游泳组和生理盐水组。游泳组纹状体多巴胺转运体相对密度较非游泳组和生理盐水组明显下调。后两组无差异。结论游泳运动能增强小鼠的运动机能,减轻MPTP的损伤效应,纹状体多巴胺转运体下调可能是其中机制之一。     

原位鼻咽癌裸鼠模型血清蛋白质组学的研究

利用我室建立的原位鼻咽癌裸鼠模型,建立了稳定性较好的鼻咽癌裸鼠血清蛋白质2-DE方法,并比较了原位鼻咽癌裸鼠血清与对照组裸鼠血清蛋白质2-DE图谱之间的差异.与正常裸鼠血清比较,原位鼻咽癌裸鼠模型组血清增加了4个蛋白质点.利用MALDI-TOF质谱技术对血清差异蛋白点进行鉴定,发现SAA-1前体(serumamyloidA-1 protein precursor)在鼻咽癌裸鼠模型组血清中明显高表达,为寻找鼻咽癌血清生物标志物提供了有利的线索.     

GDNF-Transfected Macrophages Produce Potent Neuroprotective Effects in Parkinson's Disease Mouse Model

The pathobiology of Parkinson''s disease (PD) is associated with the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) projecting to the striatum. Currently, there are no treatments that can halt or reverse the course of PD; only palliative therapies, such as replacement strategies for missing neurotransmitters, exist. Thus, the successful brain delivery of neurotrophic factors that promote neuronal survival and reverse the disease progression is crucial. We demonstrated earlier systemically administered autologous macrophages can deliver nanoformulated antioxidant, catalase, to the SNpc providing potent anti-inflammatory effects in PD mouse models. Here we evaluated genetically-modified macrophages for active targeted brain delivery of glial cell-line derived neurotropic factor (GDNF). To capitalize on the beneficial properties afforded by alternatively activated macrophages, transfected with GDNF-encoded pDNA cells were further differentiated toward regenerative M2 phenotype. A systemic administration of GDNF-expressing macrophages significantly ameliorated neurodegeneration and neuroinflammation in PD mice. Behavioral studies confirmed neuroprotective effects of the macrophage-based drug delivery system. One of the suggested mechanisms of therapeutic effects is the release of exosomes containing the expressed neurotropic factor followed by the efficient GDNF transfer to target neurons. Such formulations can serve as a new technology based on cell-mediated active delivery of therapeutic proteins that attenuate and reverse progression of PD, and ultimately provide hope for those patients who are already significantly disabled by the disease.     

Early and Late Gene Changes in MPTP Mice Model of Parkinson's Disease Employing cDNA Microarray

Recently, we reported specific brain gene expression changes in the chronic MPTP model in the late stage of degeneration, employing cDNA expression array, which indicate a domino cascade of events involved in neuronal cell death. In an attempt to elucidate early gene expression profile in the region of the substantia nigra (SN) and the striatum of acute MPTP-treated mice (3–24 h), we elected a restricted number of genes affected by the long-term MPTP treatment, and their expression was examined. Specifically, we detected alterations in the expression of genes implicated in oxidative-stress, inflammatory processes, signal transduction and glutamate toxicity. These pro-toxic genes appear to be compensated by the elevated expression in trophic factors and antioxidant defenses, which are also activated by short exposure to MPTP. The time course of these gene expression changes indicates the importance of investigating the early gene cascade of events occurring prior to late nigrostriatal dopamine neuronal cell death.     

Rasagiline Ameliorates Olfactory Deficits in an Alpha-Synuclein Mouse Model of Parkinson's Disease

Impaired olfaction is an early pre-motor symptom of Parkinson''s disease. The neuropathology underlying olfactory dysfunction in Parkinson''s disease is unknown, however α-synuclein accumulation/aggregation and altered neurogenesis might play a role. We characterized olfactory deficits in a transgenic mouse model of Parkinson''s disease expressing human wild-type α-synuclein under the control of the mouse α-synuclein promoter. Preliminary clinical observations suggest that rasagiline, a monoamine oxidase-B inhibitor, improves olfaction in Parkinson''s disease. We therefore examined whether rasagiline ameliorates olfactory deficits in this Parkinson''s disease model and investigated the role of olfactory bulb neurogenesis. α-Synuclein mice were progressively impaired in their ability to detect odors, to discriminate between odors, and exhibited alterations in short-term olfactory memory. Rasagiline treatment rescued odor detection and odor discrimination abilities. However, rasagiline did not affect short-term olfactory memory. Finally, olfactory changes were not coupled to alterations in olfactory bulb neurogenesis. We conclude that rasagiline reverses select olfactory deficits in a transgenic mouse model of Parkinson''s disease. The findings correlate with preliminary clinical observations suggesting that rasagiline ameliorates olfactory deficits in Parkinson''s disease.     

帕金森病小鼠模型行为学检测方法的比较研究

目的比较目前常用的5种帕金森病(PD)小鼠模型行为学检测方法在PD研究中的作用。方法用MPTP建立C57BL小鼠PD模型,通过行为学检测(自主活动计数、滚轴实验、游泳实验、爬杆实验、悬挂实验)、免疫组织化学和荧光分光光度法,对比5种行为学检测方法的平均数与变异系数,观察MPTP对PD小鼠模型的行为学、黑质多巴胺(DA)神经元和纹状体酪氨酸羟化酶免疫反应阳性(TH-ir)神经纤维以及纹状体DA水平的影响。结果给与MPTP后,小鼠行为学计数降低,爬杆实验未能得到检测结果,悬挂实验变异系数很高,结果有明显的偶然性,滚轴实验结果变异系数中等,平均数呈现一定的上升趋势,自主活动计数中移动与站立和游泳实验的平均数则呈现明显的下降趋势,变异系数很低,而黑质DA神经元数目减少约58%,纹状体TH-ir神经纤维密度减低,纹状体DA水平明显降低约88%,两组相比差异有显著性(P<0.01)。结论MPTP所致的C57BL小鼠的神经病理、生化改变与PD患者近似,自主活动计数和游泳实验优于其他行为学检测方法。