中脑腹侧区细胞移植对帕金森病鼠纹状体多巴胺受体敏感 …

大鼠纹状体中多巴胺受体介导的ｃ－ｆｏｓ基因的表达与多巴胺Ｄ１受体的超敏现象有关。本实验将鼠胚胎中脑腹侧区细胞植入帕金森病大鼠模型纹状体后第１２周,用免疫组化法检测阿朴吗啡诱发的ｃ－ｆｏｓ蛋白,同时取相邻切片进行酪氨酸羟化酶检测。     

胚胎黑质移植对帕金森病鼠纹状体中多巴胺受体敏感性的影响

大鼠纹状体中多巴胺受体介导的C－fos基因的表达与多巴胺D1受体的超敏现象有关。本实验在鼠胚胎黑质细胞植入帕金森病大鼠模型纹状体后第12周,用免疫组化法检测阿朴吗啡诱发的C－fos蛋白,同时取相邻切片进行酷氨酸羟化酶检测,结果经图像分析发现胚胎黑质移植,能显著减少移植侧纹状体中C－fos的表达量,说明胚胎黑质移植能够纠正多巴胶受体的超敏现象。除此之外,还发现C－fos减少的区域明显超过相邻切片酷氨酸羟化酶免疫阳性区域,表明细胞移植对超敏的多巴胺受体的影响范围大大超过了其诱发宿主残存多巴胺神经元再生的范围。     

抗帕Ⅰ号方剂对帕金森病大鼠纹状体多巴胺受体的影响

目的探讨抗帕Ⅰ号方剂对帕金森病(PD)大鼠纹状体多巴胺D1(DR1)、D2受体(DR2)表达的影响.方法 6-羟基多巴胺损毁制备偏侧PD大鼠模型,PD大鼠分为4组,分别进行抗帕Ⅰ号方剂、左旋多巴甲酯/苄丝肼、左旋多巴甲酯/苄丝肼/抗帕Ⅰ号方剂和生理盐水灌胃治疗4周,观察大鼠行为学变化;RT-PCR检测纹状体DR1、DR2受体的表达.结果抗帕Ⅰ号方剂联合左旋多巴治疗使PD大鼠产生稳定的对侧旋转行为;左旋多巴治疗后使PD大鼠在盐酸去水吗啡诱发后产生逐步增加的对侧旋转行为;联合抗帕Ⅰ号方剂及左旋多巴治疗可使盐酸去水吗啡诱发的对侧旋转次数减少.联合左旋多巴及抗帕Ⅰ号方剂治疗后损毁侧纹状体DR1 mRNA表达较对照组、抗帕Ⅰ号方剂组增强(P<0.05),而DR2 mRNA表达较对照组、抗帕Ⅰ号方剂组减弱(P<0.05).结论联合抗帕Ⅰ号方剂及左旋多巴治疗可改善PD大鼠行为学,但单独应用抗帕Ⅰ号方剂对多巴胺受体表达无明显影响,且抗帕Ⅰ号方剂无明显受体激动剂的作用.     

帕金森病鼠纹状体内羊膜移植对C-fos表达的影响

目的:观察非多巴胺能组织-羊膜细胞移植是否能纠正帕金森病(PD)鼠纹状体多巴胺(DA)受体的超敏感性。方法:在羊膜细胞植入PD大鼠模型纹状体后第12周,用免疫组化法检测阿朴吗啡诱发的C-fos蛋白。结果:经图像分析,发现成活羊膜细胞移植与死羊膜细胞移植相比,能显著减少移植侧纹状体中C-fos的表达量。结论:成活羊膜细胞移植能够纠正多巴胺受体的超敏现象。而且,多巴胺受体受影响的范围大大超过了细胞移植诱发宿主残存多巴胺神经元再生的范围。     

阿朴吗啡对颈动脉体球细胞帕金森病大鼠纹状体移植区C-FOS和JUN-B表达的影响

目的 :探讨帕金森病 (PD)大鼠颈动脉体球细胞移植治疗后多巴胺细胞的功能状况。方法 :采用 6 -羟多巴胺损毁制备 PD大鼠模型 ,腹腔注射阿朴吗啡 2 h后诱导移植后 12周纹状体组织内 c- fos和 Jun- B的表达 ,分析其分布和阳性细胞数目。结果 :移植后 12周 ,移植物内和与宿主接触面 c- fos表达增高 ,而 Jun- B的表达没有变化。结论 :移植物内和与宿主接触面 c- fos表达增高表明移植细胞仍保持着其生理功能     

异体颈动脉体球细胞脑内移植对帕金森病大鼠行为及纹状体多巴胺含量的改善

帕金森病（Parkinson disease,PD)是一种好发于中老年人的中枢神经系统退行性疾病,细胞移植治疗是一项重要的手段,近年来,有研究表明颈动脉体球细胞在低氧状态下生长良好,且具有旺盛的分泌多巴胺（DA）的特性,为此,我们利用颈动脉体球细胞作为移植组织来源,系统观察移植治疗偏侧PD大鼠行为及DA的改善情况。     

芳香族氨基酸脱羧酶转基因骨骼肌细胞脑内移植后对纹状体区多巴胺含量的影响

目的探讨帕金森病(PD)大鼠模型脑内移植人类神经元性芳香族氨基酸脱羧酶(AADC)基因对PD的治疗作用及其在左旋多巴(L-dopa)治疗过程中的地位,并检测脑内多巴胺含量的变化.方法将pCDNA3-AADC转染的原代培养的骨骼肌细胞,移植于SD大鼠PD模型毁损侧纹状体,并在此基础上予以外源性 L-dopa 10 mg·kg-1·d-1腹腔内注射,分别观察基因治疗前后以及结合给予外源性L-dopa前后各组动物病理性旋转行为的改善,并行高效液相电化学法检测脑内多巴胺(DA)含量.结果 AADC转基因骨骼肌细胞脑内移植后,实验组PD模型大鼠旋转行为较前明显改善,并可持续15周以上,尤以第11周时最为明显,约为64.6%(P<0.05),结合外源性L-dopa治疗后动物模型的旋转行为有更进一步的改善,达到86.5%;脑内DA含量测定证实,PD模型鼠脑内AADC水平较健康鼠明显降低,仅(2 119.0±47.2) ng/mg,约为健康鼠的31.0%;在补充了AADC后,脑内纹状体区DA含量则显著提高,达到(3 907.5±56.3) ng/mg;结合补充外源性L-dopa后,脑内DA的水平得到进一步提高达(5 443.0±78.7) ng/mg.结论脑内植入AADC转基因骨骼肌细胞,增加了脑内AADC基因的表达,可有效改善帕金森病大鼠的旋转行为,并可通过增加对L-dopa的脱羧作用而提高其治疗效果,有助于在低剂量上长期维持L-dopa的疗效,减轻L-dopa治疗中的不良反应.     

新型多巴胺受体激动剂与帕金森病

帕金森病(Parkinson's disease.PD)是一种常见的锥体外系疾病,临床表现以震颤、肌肉僵硬为主.伴有植物神经功能、认知和情感功能障碍等非运动症状Ⅲ.其病理改变为中脑黑质的多巴胺(dopamine.DA)能神经元缓慢进行性变性和死亡,导致黑质、纹状体DA转运体、DA显著减少.最终导致锥体外系功能失调,从而出现上述行为障碍.     

多巴胺受体激动剂治帕金森病的临床研究进展

本文对多巴胺受体激动剂的分类、作用机制、临床疗效、不良反应、应用方式以及临床上如何正确选择多巴胺受体激动剂作了系统阐述,强调了多巴胺受体激动剂在帕金森病治疗中的重要地位,尤其在疾病早期就作为单药使用的优越性。     

胚胎多巴胺神经元移植治疗帕金森病的实验研究

目的探讨胚胎多巴胺神经元移植对帕金森病大鼠的治疗作用。方法立体定向注射6-羟多巴胺建立帕金森病大鼠模型,随机分为对照组(n=12)和细胞移植组(n=12)。细胞移植组将荧光染料CM-DiI标记的大鼠胚胎多巴胺神经元立体定向注入帕金森病大鼠纹状体区,对照组于相同部位注入生理盐水。用阿扑吗啡诱导帕金森病大鼠旋转行为评估细胞移植的治疗作用。细胞移植8周后取大脑标本行冷冻切片,荧光显微镜下观察移植细胞在脑内存活情况。结果与对照组比较,移植多巴胺神经元能显著改善阿扑吗啡诱导帕金森病大鼠的异常旋转行为(P0.01)。移植8周后,仅少量多巴胺神经元存活。结论多巴胺神经元移植可短期内改善帕金森病大鼠的运动障碍,但长期疗效不佳,可能与移植多巴胺神经元长期存活率较低有关。     

急性Dieldrin中毒对C57BL小鼠纹状体多巴胺含量的影响

目的：探讨急性Ｄｉｅｌｄｒｉｎ（氧桥氯甲桥奈）中毒对Ｃ５７ＢＬ 小鼠纹状体多巴胺及其代谢产物含量的影响。方法：给３组小鼠分别喂食 Ｄｉｅｌｄｒｉｎ ５０ ｍｇ·ｋｇ－１体重、Ｄｉｅｌｄｒｉｎ ４０ ｍｇ·ｋｇ－１体重和等量的生理盐水,３ ｈ后处死小鼠,取纹状体匀浆用高效液相色谱－电化学检测法测定纹状体中多巴胺（ＤＡ）、５－羟色胺（５－ＨＴ）、高香草酸（ＨＶＡ）、５－羟基吲哚乙酸（５－ＨＩＡＡ）和去甲肾上腺素（ＮＡ）的含量。结果：喂食 Ｄｉｅｌｄｒｉｎ ５０ ｍｇ·ｋｇ－１体重能降低小鼠纹状体中 ＮＡ、ＤＡ及 ＨＶＡ的含量．结论：大剂量的Ｄｉｅｌｄｒｉｎ可影响小鼠纹状体中 ＤＡ的代谢．可能是致帕金森病的环境因素之一。     

百草枯所致帕金森病小鼠黑质纹状体通路多巴胺转运体减少

目的 :探讨多巴胺转运体 (dopaminetransporter ,DAT)是否参与百草枯所致PD的发病机制。方法 :用口服百草枯的途径 ,建立小鼠帕金森病模型 ;应用免疫组织化学和原位杂交方法分别观察小鼠纹状体区DAT的水平和黑质部基因表达的变化。结果 :每天口服百草枯 10mg·kg-1的C5 7BL/6小鼠 ,2个月后自发性活动明显减少。纹状体区的DAT含量较口服盐水对照组减少 42 % (P <0 0 1) ,黑质部DATmRNA的表达降低 3 7 2 % (P <0 0 1)。结论 :百草枯可造成小鼠帕金森病样的行为表现。黑质纹状体通路DAT含量和基因表达的降低 ,提示DAT参与了百草枯所致的帕金森病发病机制     

大鼠胚胎中脑前体细胞培养的研究

目的 :通过胚胎MPC体外培养获取PD患者CRT治疗所需的DA能神经元。方法 :取自E12鼠胚中脑腹侧的MPC悬液在添加bFGF的DMEM/F12 /N2培养液中原代培养 ,鉴定培养细胞的增殖和分化能力。结果 :在添加了bFGF 10ng·mol-1的DMEM/F12 /N2培养液中MPC增殖良好 ,体外培养 5～ 7d后细胞数量扩增到培养前的 9 782± 0 0 47倍 ;培养液中撤去bFGF后细胞可分化成星形胶质细胞和神经元 ,其中多数Tuj1抗原标记阳性的神经元同时呈TH抗原标记阳性 ,培养细胞分化为DA能神经元的比例约为2 4 3 4% (10 2 / 419)。结论 :E12鼠胚MPC原代培养是获取DA能神经元的可靠途径 ,类似的技术应用于临床可能缓解PD患者CRT治疗供体不足的矛盾。     

百草枯对小鼠纹状体区多巴胺D1受体信号转导的影响

目的观察百草枯对小鼠纹状体区多巴胺D1受体信号转导的影响.方法用口服百草枯的途径,建立小鼠帕金森病模型;应用液闪测定技术测定百草枯对小鼠纹状体区DARPP-32(Mr=32 000)磷酸化程度的影响.结果给予小鼠口服百草枯10 mg*kg-1*d-1连续4个月后,纹状体区DARPP-32的32P结合位点增加77.4%(P＜0.01).结论百草枯可能通过cAMP/PKA系统减弱体内DARPP-32的磷酸化程度,调节其对多巴胺D1受体信号转导途径的作用.     

Corticosterone Attenuates Reward-Seeking Behavior and Increases Anxiety via D2 Receptor Signaling in Ventral Tegmental Area Dopamine Neurons

Corticosteroids (CORT) have been widely used in anti-inflammatory medication. Chronic CORT treatment can cause mesocorticolimbic system dysfunctions, which are known to play a key role for the development of psychiatric disorders. The VTA is a critical site in the mesocorticolimbic pathway and is responsible for motivation and reward-seeking behaviors. However, the mechanism by which chronic CORT alters VTA dopamine neuronal activity is largely unknown. We treated periadolescent male mice with vehicle, 1 d, or 7 d CORT in the drinking water, examined behavioral impacts with light/dark box, elevated plus maze, operant chamber, and open field tests, measured the effects of CORT on VTA dopamine neuronal activity using patch-clamp electrophysiology and dopamine concentration using fast-scan cyclic voltammetry, and tested the effects of dopamine D2 receptor (D2R) blockade by intra-VTA infusion of a D2R antagonist. CORT treatment induced anxiety-like behavior as well as decreased food-seeking behaviors. We show that chronic CORT treatment decreased excitability and excitatory synaptic transmission onto VTA dopamine neurons. Furthermore, chronic CORT increased somatodendritic dopamine concentration. The D2R antagonist sulpiride restored decreased excitatory transmission and excitability of VTA dopamine neurons. Furthermore, sulpiride decreased anxiety-like behavior and rescued food-seeking behavior in mice with chronic CORT exposure. Together, 7 d CORT treatment induces anxiety-like behavior and impairs food-seeking in a mildly aversive environment. D2R signaling in the VTA might be a potential target to ameliorate chronic CORT-induced anxiety and reward-seeking deficits.SIGNIFICANCE STATEMENT With widespread anti-inflammatory effects throughout the body, corticosteroids (CORT) have been used in a variety of therapeutic conditions. However, long-term CORT treatment causes cognitive impairments and neuropsychiatric disorders. The impact of chronic CORT on the mesolimbic system has not been elucidated. Here, we demonstrate that 7 d CORT treatment increases anxiety-like behavior and attenuates food-seeking behavior in a mildly aversive environment. By elevating local dopamine concentration in the VTA, a region important for driving motivated behavior, CORT treatment suppresses excitability and synaptic transmission onto VTA dopamine neurons. Intriguingly, blockade of D2 receptor signaling in the VTA restores neuronal excitability and food-seeking and alleviates anxiety-like behaviors. Our findings provide a potential therapeutic target for CORT-induced reward deficits.     

大鼠胚胎中脑神经前体细胞培养、移植治疗大鼠PD模型

目的:研究原代培养的大鼠胚胎中脑前体细胞(mesencephalicprogenitorcells ,MPC)作为帕金森病(Parkinson’sdisease ,PD)细胞替代疗法供体的可行性。方法:PD模型大鼠19只,随机分为细胞移植组(n =9)、假移植组(n =5 )和对照组(n =5 ) ;定期测定移植前后各模型鼠阿扑吗啡诱导的旋转行为的变化,免疫组化定性供体细胞在宿主体内存活和分化的情况。结果:术后6和16周,细胞移植组阿扑吗啡诱导旋转的相对频率与术前比较有显著差异;细胞移植组术后16周时的阿扑吗啡诱导旋转的相对频率平均值与对照组对应的相对频率平均值相比也有显著差异;供体细胞能在宿主体内分化为多突起的DA能神经元。结论:原代培养的胚胎MPC可能作为PD患者细胞替代治疗的供体细胞。     

Subregion-Specific Regulation of Dopamine D1 Receptor Signaling in the Striatum: Implication for L-DOPA-Induced Dyskinesia

The striatum is the main structure of the basal ganglia. The striatum receives inputs from various cortical areas, and its subregions play distinct roles in motor and emotional functions. Recently, striatal maps based on corticostriatal connectivity and striosome-matrix compartmentalization were developed, and we were able to subdivide the striatum into seven subregions. Dopaminergic modulation of the excitability of medium spiny neurons (MSNs) is critical for striatal function. In this study, we investigated the functional properties of dopamine signaling in seven subregions of the striatum from male mice. By monitoring the phosphorylation of PKA substrates including DARPP-32 in mouse striatal slices, we identified two subregions with low D1 receptor signaling: the dorsolateral portion of the intermediate/rostral part (DL-IR) and the intermediate/caudal part (IC). Low D1 receptor signaling in the two subregions was maintained by phosphodiesterase (PDE)10A and muscarinic M4 receptors. In an animal model of 6-hydroxydopamine (6-OHDA)-induced hemi-parkinsonism, D1 receptor signaling was upregulated in almost all subregions including the DL-IR, but not in the IC. When L-DOPA-induced dyskinesia (LID) was developed, D1 receptor signaling in the IC was upregulated and correlated with the severity of LID. Our results suggest that the function of the striatum is maintained through the subregion-specific regulation of dopamine D1 receptor signaling and that the aberrant activation of D1 receptor signaling in the IC is involved in LID. Future studies focusing on D1 receptor signaling in the IC of the striatum will facilitate the development of novel therapeutics for LID.SIGNIFICANCE STATEMENT Recent progress in striatal mapping based on corticostriatal connectivity and striosome-matrix compartmentalization allowed us to subdivide the striatum into seven subregions. Analyses of D1 receptor signaling in the seven subregions identified two unique subregions with low D1 receptor signaling: the dorsolateral portion of the intermediate/rostral part (DL-IR) and the intermediate/caudal part (IC). Aberrant activation of D1 receptor signaling in the IC is involved in L-DOPA-induced dyskinesia (LID). Previous studies of LID have mainly focused on the DL-IR, but not on the IC of the striatum. Future studies to clarify aberrant D1 receptor signaling in the IC are required to develop novel therapeutics for LID.     

帕金森病患者脑脊液中抗D2受体抗体的检测

目的检测帕金森病(PD)患者脑脊液(CSF)中抗D2受体抗体.方法采用免疫组化和免疫印迹检测30例PD患者CSF,15例正常人CSF中的抗D2受体抗体.结果7例浓缩10倍的PD-CSF存在抗D2受体抗体,18例未浓缩PD-CSF及15例正常人CSF未检测到该抗体.结论部分PD患者CSF中存在抗D2受体抗体.