电针治疗前后帕金森病模型大鼠纹状体D_2受体与多巴胺含量的对比研究(摘要)

目的　研究经电针治疗前后偏侧帕金森病(PD)模型大鼠纹状体多巴胺(DA)D2受体的变化,并将其与DA及其代谢产物的含量进行对比分析。方法　用125I3碘2羟基6甲氧基N[(1乙基2吡咯烷)甲基]苯酰胺(IBZM)脑D2受体放射自显影、高效液相电化学检测器(HPLCECD)检测治疗前组、电针治疗组及治疗空白对照组偏侧PD模型大鼠纹状体DA及其代谢产物高香草酸(HVA)、3,4二羟苯乙酸(DOPAC)含量。结果　电针治疗组患侧纹状体DA、HVA、DOPAC含量较治疗前组及治疗空白对照组增高(P<0.05);电针治疗组患侧纹状体/小脑125IIBZM摄取比值为8.04±0.71,较健侧升高(29.34±4.83)%,与治疗前组的(8.09±0.52)相比,差异无显著性(P>0.05),但低于治疗空白对照组的(8.61±0.63),此值较健侧升高(38.63±3.71)%。结论　电针治疗可以提高大鼠实验性偏侧PD模型纹状体DA含量,并可抑制早期实验性PD时D2受体的上调效应。     

牛磺酸抑制帕金森病模型大鼠的旋转行为及其可能机制

探讨牛磺酸对帕金森病模型大鼠由阿朴吗啡(APO)诱发的向健侧的旋转行为的影响。应用6-羟基多巴胺(6-OHDA)制备偏侧损毁的帕金森病模型大鼠，侧脑室(icv)微量注射不同浓度的牛磺酸，1w后在自动旋转测试仪中测试由APO诱发的旋转行为，并通过高效液相色谱电化学检测(HPLC-ECD)检测纹状体中多巴胺(DA)及其代谢产物的含量。     

建立类似于绝经期妇女帕金森病大鼠模型的实验研究

目的探讨利用6-羟基多巴(6-OHDA)制备模拟绝经期妇女帕金森病的大鼠模型。方法应用6-OHDA制备OVX PD模型大鼠,应用免疫组织化学染色、高效液相色谱法等技术对大鼠黑质(SN)的酪氨酸羟化酶(TH)阳性神经元数目、纹状体多巴胺(DA)及其代谢物二羟基苯乙酸(DOPAC)和高香草酸(HVA)含量进行考察并评价。结果阿朴吗啡可诱导出明显的PD大鼠旋转行为;大鼠损伤侧黑质TH阳性神经元数量较健侧显著减少(P<0.01),纹状体DA及其代谢物DOPAC和HVA含量也较健侧明显减少(P<0.01)。结论本模型具有绝经期妇女帕金森病的基本的病理特点。     

牛磺酸对胚胎中脑细胞移植后的保护作用

①目的　观察牛磺酸对帕金森病 (PD)模型大鼠胚胎中脑细胞移植术后移植细胞的存活及生长的影响。②方法　应用 6 羟基多巴胺 (6 OHDA)制备偏侧损毁的PD模型大鼠 ,将胚胎腹侧中脑 (VM )细胞悬液或生理盐水 (NS) (对照组 )立体定位移植到模型大鼠的损毁侧纹状体中 ,VM移植大鼠于术后侧脑室分别注射NS或牛磺酸 ,5 6d后通过免疫组织化学酪氨酸羟化酶 (TH)染色观察纹状体中多巴胺 (DA)能神经元 (TH阳性细胞 )的存活及生长情况。③结果　行胚胎中脑细胞移植术后 ,在PD模型大鼠的损毁侧纹状体中可检测到移植成活的DA能神经元 ,侧脑室注射牛磺酸组比侧脑室注射NS组细胞成活数目明显增多 (F =5 0 8.5 ,q =3.9,P <0 .0 1)。而在对照组模型大鼠的损毁侧纹状体中未检测到DA能神经元。④结论　牛磺酸可有效改善移植细胞的存活条件 ,并促进移植细胞的生长。     

MPTP对小鼠纹状体多巴胺含量的影响

报道1—甲基—4—苯基—1,2,3,6—四氢吡啶(MPTP)给C_(57)-BL黑色小鼠腹腔注射后,应用高效液相色谱—电化学检测法,测定纹状体多巴胺(DA)、去甲肾上腺素(NA)、5—羟色胺(5-HT)及其代谢产物3,4—二羟基苯乙酸(DOPAC)、高香草酸(HVA),5—羟基吲哚乙酸(5-HIAA)的含量。结果与对照组相比,用药组小鼠纹状体DA、5-HT及其代谢产物显著降低,其中DA减少94%(P<0.001),DOPAC减少61%(P<0.05),HVA减少65%(P<0.001)。实验结果表明用MPTP给予小鼠所产生的动物模型,对研究帕金森病是一种较为有用的方法。     

黑质致密部和内侧前脑束注射6-OHDA 制备的帕金森病大鼠模型纹状体中 DA 含量比较

目的：：比较黑质致密部(SNc)损毁和内侧前脑束(MFB)损毁2种方法制备的帕金森病(PD)大鼠模型对纹状体中多巴胺(DA)递质含量的影响。方法：将大鼠随机分为假手术组(n=12)、SNc 损毁组(n=15)和 MFB 损毁组(n=14)。采用高效液相色谱-电化学检测法,观察3组大鼠损毁侧纹状体中 DA的含量。结果：与假手术组相比较,SNc 损毁组(P <0.001)和 MFB 损毁组(P <0.001)大鼠纹状体中 DA含量均显著降低,与 SNc 损毁组相比较,MFB 损毁组大鼠纹状体中 DA 含量下降更为显著(P =0.005)。结论：MFB 损毁制备的 PD 大鼠模型对 DA 能神经元的损伤范围较 SNc 损毁有所扩大,为不同研究选择制备模型的方法提供一定的理论依据。     

黑质内注射FeCl3对大鼠纹状体多巴胺释放量及含量的影响

①目的探讨黑质(SN)内注射不同剂量的FeCl3对大鼠纹状体(CPu)多巴胺(DA)释放量和含量的影响.②方法实验用SD大鼠32只,分成4组6只大鼠为正常对照组;9只大鼠左侧SN内注射 FeCl3溶液10 μg(Ⅰ组);8只大鼠左侧SN内注射FeCl3溶液20 μg(Ⅱ组);9只大鼠左侧SN内注射FeCl3溶液40 μg(Ⅲ组),3周后采用快速周期伏安法(FCV)监测纹状体D A的释放量,高效液相色谱法(HPLC)检测纹状体DA含量.③结果在FeCl3单侧损毁大鼠中,损毁侧CPu DA释放量的减少和DA含量的降低与注射FeCl3剂量之间有明显剂量依赖关系(F=93.78,164.51,q=3.50～29.79,P<0.05).Ⅲ组损毁侧CP u DA的更新率[以(DOPAC+HVA)/DA表示]明显加快,与其他组比较差异有显著性(F=13 6.22,q=22.32～22.72,P<0.05);而Ⅰ,Ⅱ组变化较小,与正常大鼠相比差别无统计学意义 (q=0.25,2.04,P>0.05).④结论黑质内Fe3+含量升高可使CPu内DA释放量和含量均降低,铁含量高可能参与帕金森病的病理改变.     

PD大鼠健侧纹状体多巴胺能系统的功能性改变

应用快速周期伏安法(FCV)在体监测电刺激内侧前脑束(MFB)诱发的正常和帕金森病(PD)大鼠健侧及损毁侧纹状体(Str)内多巴胺(DA)的释放，并结合高效液相色谱电化学检测法(HPLC-ECD)测定Str内DA及其代谢产物的含量．从在体和离体水平分别对PD大鼠健侧及损毁侧Str区DA的释放及代谢进行观察及评价。实验采用自身对照，先用FCV监测DA的释放，后行HPLC-ECD离体测定Str内DA及其代谢产物的含量。     

褪黑激素对实验性帕金森病大鼠行为及大脑多巴胺代谢的影响

目的：观察褪黑激素（Melatonin）对MPTP模型大鼠行为学和前脑多巴胺及其代谢产物含量的影响，探讨褪黑激素治疗帕金森病（PD）的作用及其机制。方法：SD大鼠60只，分为空白对照组、病理对照组和用药实验组Ⅰ～Ⅲ5组。病理对照组和用药实验组注射MPTP建立PD大鼠模型，采用爬杆法和迷宫试验测量大鼠行为学和智力改变，应用高压液相电化学法测定前脑多巴胺（DA）、高香草酸（HVA）及二羟基苯乙酸（DOPAC）含量。结果：PD模型大鼠出现不同程度爬杆能力下降和智力减退，与病理对照组相比用药实验组大鼠行为学异常可部分或全部改善。病理对照组DA、HVA、DOPAC含量均较空白对照组显著降低，用药实验组DA、HVA、DOPAC含量较空白对照组低，但较病理对照组明显增加。结论：褪黑激素可能通过抗氧化应激途径，调节前脑DA神经活性及其代谢，以改善PD症状。     

底丘脑核中MPEP对治疗早期帕金森病模型大鼠运动行为的影响

目的：：探讨底丘脑核(STN)中代谢型谷氨酸受体5(mGluR5)拮抗剂 MPEP 对部分损毁的帕金森病(PD)模型大鼠运动行为的影响及神经保护作用。方法：将大鼠随机分为假手术组(n=12)、损毁+生理盐水组(n=12)、损毁+MPEP 组(n=12)。采用行为学和神经化学检测方法,观察3组大鼠在旷场实验中的自发运动能力和斜板实验中的停留角度,以及纹状体中多巴胺(DA)含量。结果：与假手术组大鼠相比较,损毁+生理盐水组大鼠的水平运动能力(P<0.001)、垂直运动能力(P<0.001)和斜板停留角度(P<0.001)均明显降低,而损毁+MPEP组大鼠均恢复至假手术组水平(P>0.05)。神经化学检测结果显示,与假手术组大鼠相比较,损毁+生理盐水组大鼠纹状体DA含量降低了56.3%(P<0.001),而损毁+MPEP组大鼠与假手术组大鼠无明显差异(P>0.05),且与损毁+生理盐水组比较明显升高(P<0.001)。结论：慢性STN局部应用MPEP对早期PD模型大鼠的运动行为和神经元具有保护作用。     

锰卟啉络合物对MPTP诱导的帕金森病小鼠的防治作用

目的:观察锰卟啉络合物[manganese(Ⅲ)meso-tetrakis(N,N'-diethylimidazolium-2-yl) porphyrin,MnTDM]对1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)诱导的早期帕金森病模型小鼠的防治效果,探讨其可能的作用机制.方法:C57BL/6雄性小鼠随机分为MPTP模型组(连续3 d皮下注射25 mg/kg MPTP),MnTDM MPTP组(于MPTP注射前1 h皮下注射15 mg/kg MnTDM)以及MnTDM对照组、生理盐水对照组,每组10只.末次注射后第3日进行爬杆和游泳等行为学检测;HPLC-ECD法检测各组小鼠纹状体多巴胺(dopamine,DA)及其代谢产物3,4-二羟基苯乙酸(DOPAC)和高香草酸(HVA)水平;硫代巴比妥酸(thiobarbituric acid,TBA)法测定各组小鼠纹状体丙二醛(malondialdehyde,MDA)水平.结果:急性注射MPTP可建立早期帕金森病小鼠模型;与对照组相比,MPTP组小鼠纹状体DA、DOPAC、HVA水平明显下降(P＜0.01),MDA水平明显升高(P＜0.05);但短期对小鼠行为学指标影响不大.MnTDM能部分抑制MPTP的上述作用;与MPTP组相比,MnTDM MPTP组小鼠纹状体DA、DOPAC、HVA水平明显上升,MDA水平明显下降(P均＜0.05).各组小鼠间行为学指标无统计学差异.结论:MnTDM能抑制脂质过氧化,促进多巴胺类神经递质分泌,对MPTP诱导的帕金森病小鼠有一定防治作用.     

Effects of Echinacoside on Histio-central Levels of Active Mass in Middle Cerebral Artery Occlusion Rats*

Objective To investigate the effects of echinacoside on the extracellular striatal levels of norepinephrine (NE),dopamine (DA),homovanillic acid (HVA),3,4-dihydroxyphenylethanoid acid (DOPAC),5-hydroxyindoleacetic acid (HIAA),and 5-hydroxytryptamine(5-HT) in middle cerebral artery occlusion (MCAO rats.Methods The middle cerebral artery was occluded in male Sprague-Dawley rats.Three days later microdialysis probes were placed into the right striatum of MCAO rat brains and the brains were perfused with Ringer's solution at a rate of 1.5 μL/min.Cerebral microdialysates were collected every 30 minutes from awake and freely moving rats before assaying for NE,DA,HVA,DOPAC,HIAA,and 5-HT levels by reverse phase HPLC with electrochemistry.Results Three days after MCAO,the extracellular striatal levels of NE,DA,DOPAC,HIAA,HVA,and 5-HT of the MCAO rats increased significantly (at least P＜0.05 vs.control).However,simultaneous treatment with echinacoside (30.0 or 15.0 mg/kg) attenuated these increases (at least P＜0.05 vs.non-treated model rats).Conclusion These results imply that echinacoside may protect striatal dopa minergic neurons from the injury induced by MCAO and may help prevent and treat cerebral ischemic diseases.     

PROTECTIVE ACTION OF BUDIPINE AGAINST MPTP NEUROTOXICITY

Mice treated with large dose of MPTP showed a reduction in the levels of DA, 5-HT and their metabolites in the striatum. The average levels as compared with the control were reduced by 94% for DA (P<0.001), 61% for DOPAC (P<0.05), 65% for HVA (P<0.01). 5-HT and 5-HIAA were lower than detection limits. In mice pretreated with budipine, although the striatal dopamine level was also reduced, mean DA and 5-HT levels were significantly higher than those in mice given MPTP alone. This result suggested that budipine can partially prevent the MPTP neurotoxicity.     

葛根素对急性乙醇中毒大鼠脑内阿片肽和多巴胺的调控作用

目的： 检测葛根素对急性乙醇中毒大鼠皮层、小脑、海马和纹状体内阿片肽和多巴胺（DA）表达水平的调控作用,为临床防治乙醇中毒和开发中药葛根提供依据。方法： 雄性SD大鼠24只随机分为对照组、乙醇中毒组和葛根素组,每组8只,采用放射免疫法检测各组大鼠皮层、小脑、海马和纹状体内β-内啡肽（β-EP）、强啡肽（DnyA）和亮脑啡肽（L-EK）表达水平,采用高效液相色谱法检测各组大鼠各脑区DA、多巴克（DOPAC）和高香草酸（HVA）的表达水平,并计算（DOPAC+HVA）/DA的比值。结果：与对照组比较,乙醇中毒组和葛根素组大鼠皮层内β-EP、DnyA和L-EK表达水平降低(P<0.01),皮层、小脑、海马和纹状体内 DA、DOPAC和HVA表达水平均升高 (P<0.01);与乙醇中毒组比较,葛根素组大鼠皮层内β-EP、DnyA和L-EK表达水平升高(P<0.05,P<0.01),DA(P<0.01)、DOPAC(P<0.01)和HVA表达水平均降低,小脑、海马和纹状体内所有指标表达水平均降低(P<0.05,P<0.01),皮层、小脑和海马内（DOPAC+HVA）/DA 比值升高,纹状体内该比值降低（P>0.05）。结论：葛根素对急性乙醇中毒造成的脑组织损伤有保护作用,其机制可能与葛根素对脑内阿片肽和DA的调控有关。     

震颤宁治疗帕金森病的实验研究

[目的]探讨中药襄颤宁治疗帕金森氏病(PD)的药效机理。[方法]采用6-羟多巴胺(6-OHDA)纹状体两点注射法复制PD模型,观察震颤宁对大鼠旋转行为、纹状体及血液中多巴胺(DA)和高香草酸(HVA)、过氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-PX)含量的影响。[结果]高剂量震颤宁可改善PD大鼠旋转行为,使旋转圈数较治疗前减少(P<0.05);提高大鼠全血GSH舍量(P<0.05),使GSH-PX活性显著增高(P<0.01),血清MDA含量显著降低:右侧纹状体GSH含量显著增高(P<0.01),MDA含量显著降低(P<0.01),并可增加纹状体DA含量,降低HVA/DA比值。低剂量震颤宁对PD大鼠旋转行为无明显改善(P>0.05);可降低血清MDA含量(P<0.01),而对GSH-PX活性无明显改善;并可使大鼠纹状体GSH含量增加,而对MDA含量无明显影响,对DA、HVA/DA亦无显著影响。[结论]震颤宁可能通过改善机体及纹状体的氧化应激状态和保护神经元,间接补充纹状体DA含量而改善PD症状,并呈剂量效应关系。     

Comparative study of D2 receptors and dopamine content in striatum before and after electro-acupuncture treatment in rats

Objective To evaluate the change in D(2) receptors and their relationship with dopamine (DA) content in experimental hemi-parkinsonism rats before and after electro-acupuncture (EA) treatment. Methods (125) Ⅰ-IBZM D(2) receptor cerebral autoradio-graphic analysis, HPLC-ECD detection of DA and its metabolites, homovanillic acid (HVA), 3,4-dihydroxyphenylacetic acid (DOPAC) were used to study their levels in striatum in pre-EA, EA and EA control group. Results The DA, HVA and DOPAC levels in striatum of the lesioned side in the EA group were elevated compared to the pre-EA and EA control group (P&lt;0.05). For the EA group, the striatum/cerebellum (125) Ⅰ-IBZM uptake ratio of the lesioned side was 8.04±0.71, (29.34%±4.83%) more than that of the contralateral side (P&lt;0.05), while no significant difference was observed as compared with that in the pre-EA group (8.09±0.52, 30.12%±4.53%, higher than that of the intact side P&gt;0.05). It was lower than the EA control group (8.61±0.63, P&lt;0.05), and the latter was (38.63%±3.71%) higher than that in its contralateral side (P&lt;0.05). Conclusion 6-OH-DA lesions in the substantia nigra and ventral tegmental areas induce an up-regulation of striatal D(2) binding sites. EA treatment could elevate the DA level of the lesioned side striatum and prevent D(2) receptor up-regulation in rats with experimental hemi-parkinsonism.     

Therapeutic effect of human amniotic epithelial cell transplantation into the lateral ventricle of hemiparkinsonian rats

Background Human amniotic epithelial cells （HAECs） are able to secrete biologically active neurotrophins such as brain-derived neurotrophic factor and neurotrophin-3, both of which exhibit trophic activities on dopamine neurons. Previous study showed that when human amniotic epithelial cells were transplanted into the striatum of 6-hydroxydopamine （6-OHDA）-induced Parkinson disease rats, the cells could survive and exert functional effects. The purpose of this study was to investigate the survival and the differentiation of human amniotic epithelial cells after being transplanted into the lateral ventricle of Parkinson＇s disease （PD） rats, and to investigate the effects of grafts on healing PD in models. Methods The Parkinson＇s model was made with stereotactic microinjection of 6-hydroxydopamine （6-OHDA） into the striatum of a rat. The PD models were divided into two groups： the HAECs group and the normal saline （NS） group. Some untreated rats were taken as the control. The rotational asymmetry induced by apomorphine of the HAECs group and the NS group were measured post cell transplantation. The expression of nestin and vimentin in grafts were determined by immunohistology. Ten weeks after transplantation the density of tyrosine hydroxylase positive cells in the substantia nigra of the HAECs group, NS group and the untreated group was determined. The differentiation of grafts was determined by TH immunohistology. High performance liquid chromatography （HPLC） was used to determine monoamine neurotransmitter levels in the striatum. Results The rotational asymmetry induced by apomorphine of the HAECs group was ameliorated significantly compared to the NS group two weeks after transplantation （P 〈0.01）. The grafts expressed nestin and vimentin five weeks after transplantation. TH immunohistochemistry indicated that the TH positive cells in the substantia nigra of the HAECs group increased significantly compared to the NS group （P 〈0.01）. Tyrosine hydroxylase （TH） positive cells in the substantia nigra of the HAEC group and the NS group were decreased compared to the untreated group （P 〈0.01）. Dopamine and DOPAC levels in the striatum of the HAECs group increased significantly compared to the NS group （P 〈0.05）. Homovanillic acid （HVA） levels in the striatum of the HAECs group increased significantly compared to the NS group （P 〈0.01）. In addition dopamine, DOPAC, and HVA levels in the striatum and dopamine levels in the cerebrospinal fluid of the HAECs group and the NS group were decreased compared to the untreated group （P 〈0.05）. Conclusions Human amniotic epithelial cells could be used to ameliorate the rotational asymmetry induced by apomorphine of the PD models. This could have been due to the increased content of dopamine and its metabolic products, DOPAC and HVA, in the striatum in the PD models.     

姜黄素类似物L6H3对帕金森病大鼠脑部纹状体中单胺类神经递质代谢物的影响

目的：研究姜黄素类似物L6H3对帕金森病（PD）大鼠脑纹状体中7种单胺类神经递质及其代谢物的影响。方法：采用脑部纹状体定位注射6-羟基多巴胺（6-OHDA）建立PD大鼠模型,将大鼠随机分为假手术组、PD组和PD+L6H3组。通过微透析活体取样技术和高效液相色谱电化学检测法（HPLC-ECD）联用测定PD组大鼠脑纹状体细胞外液中单胺类神经递质及其代谢物含量。结果：造模后第6周（即给药第2周）及造模后第7周（即停药第1周）,PD+L6H3组大鼠脑纹状体的多巴胺（DA）、3,4-二羟基苯乙酸（DOPAC）、高香草酸（HVA）、去甲肾上腺素（NE）、肾上腺素（E）、5-羟色胺（5-HT）、5-羟基吲哚乙酸（5-HIAA）与PD组比较明显升高（P＜0.01）。结论：L6H3在给药后第2周及停药后1周均能明显提高PD大鼠脑中单胺类神经递质及其代谢物的含量。     

3 种不同脑区定点注射 6-OHDA 单侧损伤大鼠帕金森病模型的比较研究

[摘要] 目 的 从 神 经 行 为 学、组 织 病 理 和 生 化 方 面 对 3 种 不 同 脑 区 单 侧 定 点 注 射 6- 羟 基 多 巴 胺 ( 6- hydroxydopamine , 6-OHDA )损伤大鼠帕金森病( Parkinsondisease , PD )模型进行比较研究。方法 健康雄性 SD 大 鼠随机分为 3 组( n =10 ):纹状体组、黑质组、黑质 + 中脑腹侧被盖区组。根据脑立体定位图谱,将微量 6-OHDA 单点定位注入大鼠中脑黑质区、纹状体区和双点定位注入黑质区与中脑腹侧被盖区。观察阿朴吗啡诱发大鼠旋转 行为,免疫组织化学检测大鼠黑质区酪氨酸( tyrosinehydroxy-lase , TH + )阳性神经元数目,电化学高效液相色谱法 检测纹状体中多巴胺( dopamine , DA )及其代谢产物 3 , 4- 二羟苯乙酸( dihydroxy-phenylaceticacid , DOPAC )和高香 草酸( homovanillicacid , HVA )含量。结果 纹状体组 3 周后模型稳定,黑质组与黑质 + 中脑腹侧被盖区组大鼠 2 周后稳定, 3 组 30min 内向健侧转圈数均呈升高趋势,黑质 + 中脑腹侧被盖区组高于纹状体组和黑质组,组间· 时点间交互作用差异有统计学意义( P <0.05 )。纹状体组和黑质 + 中脑腹侧被盖区组造模成功率高于黑质组 ( P <0.05 )。 3 组大鼠注射 6-OHDA 损伤侧纹状体中 DA 、 DOPAC 、 HVA 含量和 TH + 神经元数目均明显低于非 损伤侧( P <0.05 )。结论 纹状体单点注射 6-OHDA 损伤建立 PD 模型成功率高,可作为研究 PD 的一种稳定可靠 的动物模型。