异动症大鼠模型基底节DARPP-32蛋白磷酸化状态的变化

目的观察左旋多巴诱发异动症(LID)大鼠模型行为学特点及DARPP-32蛋白的磷酸化状态的变化,探讨LID的发生机制。方法复制成功的帕金森病(Parkinson disease PD)大鼠应用左旋多巴治疗28d诱发LID大鼠模型,进行异常不自主运动(abnormalinvol untary movement,AIM)评分,并采用免疫印迹技术检测LID大鼠纹状体内DARPP-32蛋白Thr-34位点磷酸化水平。结果LID大鼠模型复制成功后出现了与人类LID相似的对侧上肢、躯干和口面部异常不自主运动(AIM),并随左旋多巴治疗时间的延长而加重。LID组大鼠纹状体内Thr-34位点磷酸化的DARPP-32水平较对照组与左旋多巴治疗组明显增高,又以重度LID组大鼠更为显著,差异有显著性意义(P<0.01)。结论慢性间断性给PD大鼠左旋多巴能复制出LID大鼠模型,其纹状体区DARPP-32蛋白的磷酸化状态发生了改变,与LID的发生可能有关。     

苍白球多巴胺D1样受体对帕金森病僵直症状的影响

目的探讨苍白球多巴胺D1样受体对氟哌啶醇所致帕金森病大鼠僵直症状的影响。方法大鼠单侧苍白球埋置套管,恢复3 d后腹腔注射氟哌啶醇(1 mg/kg),分别在苍白球内微量注射多巴胺D1样受体激动剂SKF38393(5 mmol/L)、D1样受体阻断剂SCH23390(5 mmol/L)以及它们的混合物,观察对氟哌啶醇导致的大鼠僵直症状的影响。结果单侧苍白球微量注射SKF38393可致帕金森病僵直模型大鼠向对侧偏转,联合给予SCH23390可阻断SKF38393所致的偏转现象。结论激活苍白球多巴胺D1样受体可减轻氟哌啶醇所致大鼠僵直症状。     

电刺激大鼠腹侧被盖区建立精神分裂症动物模型

目的建立精神分裂症的动物模型,探讨腹侧被盖区(VTA)在精神分裂症发病中的作用。方法电刺激大鼠VTA,并给予不同剂量的多巴胺受体拮抗剂氟哌啶醇(HAL)和SCH23390及促多巴胺释放剂苯丙胺,观察它们对大鼠精神分裂症模型的影响。结果刺激VTA可引发大鼠的异常行为,HAL可提高VTA刺激阈值,降低其行为评分;SCH23390可提高大鼠VTA刺激阈值;苯丙胺可降低大鼠VTA刺激阈值,差异均有显著意义(t=3.81~7.50,u=3.0~10.5,P<0.05、0.01)。结论该模型可以用来研究精神分裂症的病因及机制。     

药物干预对帕金森病大鼠行为的影响

目的通过建立大鼠帕金森病(PD)异动症(LID)模型,观察大鼠在药物干预前后的行为改变。方法选取内侧前脑束(MFB)注射6-OHDA建立单侧毁损PD鼠模型,给予左旋多巴(L-dopa 20 mg.kg-1.d-1)及生理盐水(NS,2 ml)干预满1个月,并设正常对照。观察大鼠的行为并每周1次阿朴吗啡诱发旋转行为,每2两次异常不自主运动(AIM)评分。结果MFB注射点造模成功率为75%,PD大鼠的行为改变有一定的规律性,其旋转速度存在先升后降的现象;AIM评分显示异动症发生率为72%。结论选择MFB注射点造模成功率高;PD大鼠行为变化有规律性;LID的发生与波动用药、损伤程度密切相关。     

AMPA受体分布对帕金森病及异动症发生的影响

目的观察α-氨基-3-羟基-5-甲基-4-异唑丙酸(AMPA)受体各亚型在帕金森病(PD)及异动症[左旋多巴诱发异动症(LID)]大鼠模型纹状体中亚细胞分布的变化。方法应用6-羟基多巴(6-OHDA)制备PD大鼠模型,向PD大鼠腹腔注射左旋多巴建立LID大鼠模型。引入受体蛋白质交联方法,通过Western blotting检测大鼠纹状体神经元AMPA受体各亚型在细胞表面和细胞内池不同部位含量变化。结果在PD大鼠纹状体中,AMPA受体各亚型均未发生亚细胞分布改变;慢性左旋多巴刺激诱导大鼠LID对其纹状体AMPA总蛋白以及在细胞表面和细胞内池的分布无明显影响。结论 AMPA受体分布变化对PD或LID的发生无明显影响。     

用脉冲伏安法观察多巴胺D_1D_2受体拮抗剂对大鼠纹体中DOPAC的影响

本文用脉冲伏安法在体测定麻醉大鼠纹体中二羟苯乙酸(DOPAC)的含量,并观察多巴胺受体拮抗剂氟哌啶醇(D_2/D_1)、氯丙嗪(D_2/D_1),舒必利(D_2)以及SCH23390(D_1)对纹体中DOPAC的影响。结果表明这些多巴胺受体拮抗剂均可使麻醉大鼠纹体中DOPAC含量增加,与基础DOPAC浓度(14±2μmol/L)相比,氟哌啶醇(0.5mg/kg s.c.),氯丙嗪(5mg/kgs.c.)、舒必利(5mg/kg s.c),SCH23390(1mg/kg s.c.)分别使DOPAC增加140％、102％、40％以及26％。从而证实不仅突触前D_2受体参与反馈调节DA的合成代谢,而且D_1受体也参与以上反馈调节机制,但D_2受体的调节作用较强。     

左旋多巴诱发帕金森病大鼠异动症模型的建立和评价

目的探讨左旋多巴诱发帕金森病(PD)大鼠异动症模型的建立,并从行为学角度对模型进行评价.方法6-羟多巴立体定向注射至SD大鼠(n=25)前脑内侧束,并于3周后腹腔注射阿朴吗啡(每千克体质量0.5 mg),10 min后开始计数旋转频率(共30 min),若旋转频率＞7次/分,即PD大鼠模型建立成功.采用左旋多巴甲酯(每千克体质量25 mg)腹腔注射,治疗21d,2次/天,观察大鼠异常不自主运动(AIM)的情况.结果成功模型的PD大鼠共10只,经左旋多巴治疗后,其中8只大鼠出现了AIM,包括刻板动作和对侧旋转行为,且程度各不相同;AIM积分随治疗时间的延长,呈现逐渐升高的趋势.结论左旋多巴治疗诱发的PD大鼠AIM与PD患者左旋多巴诱发异动症在某些方面相似,为深入研究异动症的机制奠定了基础.     

大鼠纹状体区多巴胺受体的变化对动物行为的影响

目的研究大鼠黑质区注射脂多糖(LPS)后,纹状体区多巴胺2型受体(D2R)亚型mRNA含量的变化,初步探讨D2R与帕金森病(PD)发病的关系.方法利用立体定向技术将LPS注入大鼠单侧黑质区,注射后21 d左右,以阿扑吗啡诱导大鼠旋转,RT-PCR方法半定量检测各组大鼠双侧纹状体区D2R两亚型mRNA的表达量.结果单侧黑质注射LPS的大鼠经阿扑吗啡诱导后向损毁同侧旋转,实验组大鼠纹状体区DL和DS的mRNA表达量损毁侧较对照侧明显增高(P＜0.05,P＜0.01).结论LPS损毁大鼠的纹状体部位D2R mRNA含量的变化,可导致大鼠行为的改变,提示D2R可能与PD的发病有一定关系.     

滋补肝肾、通络解毒中药对异动症大鼠行为学及纹状体多巴胺D2受体活性的影响

目的：观察滋补肝肾、通络解毒中药对异动症大鼠行为学和多巴胺D2受体活性的影响。方法：采用6-羟基多巴胺（6-hydroxydopamine,6-OHDA）注射于大鼠脑右侧黑质造成偏侧帕金森病（Parkinson＇s disease,PD）模型,进一步对PD大鼠予以左旋多巴/苄丝肼制作异动症（levodopa-induced dyskinesias,LID）模型。实验设立正常对照组、模型组、中药干预组、中止给药对照组和中止给药＋中药干预组,观察中药对LID大鼠异常不自主运动（abnormal involuntary movement,AIM）评分的影响,测定大鼠纹状体多巴胺D2受体的最大结合容量（maximum binding capacity,Bmax）和平衡解离常数（equilibrium dissociation constant,KD）,来评价中药对LID大鼠多巴胺D2受体亲和力的影响。结果：与模型组和中止给药对照组比较,中药干预组可明显减少异动症大鼠的AIM积分（P〈0.01）;纹状体多巴胺D2受体亲和力检查示,中药可使Bmax显著上调（P〈0.05,P〈0.01）,KD值下降（P〈0.01）,多巴胺D2受体亲和力显著提高。结论：滋补肝肾、通络解毒中药可以有效缓解异动症症状,明显改善纹状体多巴胺D2受体活性。     

滋补肝肾、通络解毒中药对异动症大鼠行为学及纹状体多巴胺D2受体活性的影响

目的：观察滋补肝肾、通络解毒中药对异动症大鼠行为学和多巴胺D2受体活性的影响。方法：采用6-羟基多巴胺（6-hydroxydopamine,6-OHDA）注射于大鼠脑右侧黑质造成偏侧帕金森病（Parkinson＇s disease,PD）模型,进一步对PD大鼠予以左旋多巴/苄丝肼制作异动症（levodopa-induced dyskinesias,LID）模型。实验设立正常对照组、模型组、中药干预组、中止给药对照组和中止给药＋中药干预组,观察中药对LID大鼠异常不自主运动（abnormal involuntary movement,AIM）评分的影响,测定大鼠纹状体多巴胺D2受体的最大结合容量（maximum binding capacity,Bmax）和平衡解离常数（equilibrium dissociation constant,KD）,来评价中药对LID大鼠多巴胺D2受体亲和力的影响。结果：与模型组和中止给药对照组比较,中药干预组可明显减少异动症大鼠的AIM积分（P〈0.01）;纹状体多巴胺D2受体亲和力检查示,中药可使Bmax显著上调（P〈0.05,P〈0.01）,KD值下降（P〈0.01）,多巴胺D2受体亲和力显著提高。结论：滋补肝肾、通络解毒中药可以有效缓解异动症症状,明显改善纹状体多巴胺D2受体活性。     

Effect of Antisense FosB and CREB on the Expression of Prodynorphin Gene in Rats with Levodopa-induced Dyskinesias

The effects of antisense FosB and CREB intra-striatum injection on the expression of prodynorphin (PDyn) gene in striatal neurons of Levodopa-induced dyskinesias (LID) rats with Parkinson disease (PD) were explored. PD model in rats was established by 6-OHDA microinjection stereotaxically. The rats were treated with chronic intermittent Levodopa celiac injection for 28 days to get the LED rats. Antisense FosB and cAMP response element-binding protein (CREB) were injected into striatum of all rats respectively. In situ hybridization was used to measure the changes in the expression of PDyn mRNA in striatum and behavior changes were observed. The results showed after administration of antisense FosB, abnormal involuntary movement (AIM) was decreased and the expression of PDyn mRNA in striatum was increased in LID rats as compared with sense FosB group (P<0.01, respectively). As compared with the control group, the expression of PDyn mRNA in striatum was decreased by antisense CREB-treated LID group (P<0.01) and compared with sense CREB treated LID group, antisense CREB-treated LID group showed no changes in AIM scores and the expressions of PDyn mRNA (both P>0.05). In conclusion, FosB protein, which replaced the CREG, could regulate the expression of PDyn mRNA and play critical role in the pathogenesis of LID.     

多巴胺受体在可卡因诱导的转录因子CREB活化中的作用

目的 研究多巴胺受体在可卡因诱导的转录因子CREB磷酸化活化中的调控作用。方法 采用D1和D3多巴胺受体抑制剂，应用Westem blotting检测D1与D3多巴胺受体在可卡因诱导的cAMP反应元件结合蛋白（CREB）磷酸化活化中的作用及D1和D3多巴胺受体抑制剂本身对CREB磷酸化活化的影响，进一步应用Western blotting检测细胞外信号调节激酶（ERK）在CREB磷酸化活化中的作用。结果 D1多巴胺受体抑制剂阻止可卡因诱导的CREB磷酸化活化，而D3多巴胺受体抑制剂促进可卡因诱导的CREB磷酸化活化，D1和D3多巴胺受体抑制剂本身不能诱导CREB磷酸化活化。MEK的特异性抑制剂SL327可以抑制可卡因诱导的CREB磷酸化活化。结论 D1和D3多巴胺受体对CREB的磷酸化活化起反式调控作用，并且这种反式调控作用依赖于ERK信号通路。     

多发性抽动症大鼠皮质及纹状体强啡肽表达特点及健脾止动汤干预作用

目的研究多发性抽动症(TS)模型大鼠皮质及纹状体区内源性阿片肽——强啡肽(Dyn)及其前体物质强啡肽原(PDyn)的表达情况,以及中药组方健脾止动汤对二者的干预作用。方法 60只健康雄性SD大鼠,随机分为造模组(45只)和空白对照组(15只),造模组采用亚氨基二丙腈(IDPN)腹腔注射法建立TS动物模型。造模成功后的动物纳入实验,随机分为模型组、泰必利组、健脾止动汤组,每组15只。模型组和空白对照组给予生理盐水灌胃,其余2组分别给予等体积的泰必利混悬液(21 mg/kg)和健脾止动汤颗粒溶液(16 g/kg),连续给药6周,每周末次给药后记录TS大鼠刻板行为积分情况。6周给药结束后,采用酶联免疫吸附法(ELISA)和实时定量荧光PCR(RT-PCR)分别检测大鼠皮质区、纹状体区Dyn和PDyn表达。结果 (1)泰必利组和健脾止动汤组TS大鼠刻板行为积分均值呈下降趋势,末次观察结果显示,2组积分均低于模型组(P0.05),其中健脾止动汤组积分低于泰必利组(P0.05);(2)组间比较,皮质区Dyn含量和PDyn mRNA相对表达量无显著差异性(P0.05);与空白对照组比较,模型组纹状体区Dyn含量和PDyn mRNA相对表达量显著升高(P0.05);给药后,泰必利组和健脾止动汤组Dyn含量和PDyn mRNA相对表达量低于模型组(P0.05),2组与空白对照组之间无显著差异性(P0.05)。结论纹状体区Dyn和PDyn表达增加可能参与TS大鼠刻板行为发生,健脾止动汤抗抽动机制可能与抑制纹状体区Dyn过表达、降低直接通路的易化作用有关。     

Effects of Ningdong granule on DA,DRD2,and HVA in a rat model of Tourette’s syndrome

OBJECTIVE:Ningdong granule is a traditional Chinese medicine preparation for the treatment of Tourette’s syndrome.METHODS:Sixty-four rats were randomly assigned to a control group and three experimental groups,respectively.Rat models of Tourette’s syndrome were established via intraperitoneal injection of apomorphine(Apo).The rats in the experimental groups were subsequently intragastrically injected with haloperidol at 10 mg/kg(haloperidol group),Ningdong granule at 370 mg/kg(NDG group),and normal saline(0.9%) at 10 mL/kg(Apo group),respectively.Rat behaviors were observed and recorded on a daily basis.After 12 w,all rats were sacrificed,and sera and striatal tissues were harvested.Homovanillic acid levels in sera,as well as dopamine and dopamine D2 receptor mRNA expression in the striatum,were measured to determine possible mechanisms of Ningdong granule on the dopamine system in a rat model of Tourette’s syndrome.RESULTS:Following intervention,stereotype actions of the Tourette’s syndrome rats were significantly inhibited in the haloperidol and NDG groups,respectively(P<0.01).Homovanillic levels were significantly greater in the haloperidol and NDG groups,respectively(P<0.05).In addition,dopamine levels were significantly less in the NDG group(P<0.01),and DRD2 mRNA expression was significantly reduced in the haloperidol and NDG groups,respectively(P<0.05).CONCLUSION:Results demonstrated that Ningdong granule effectively inhibited stereotype actions and Tourette’s syndrome symptoms by promoting dopamine metabolism,reducing dopamine levels in the striatum,increasing homovanillic acid content in sera,and reducing mRNA expression of DRD2 in the striatum.     

用脉冲伏安法观察多巴胺D1D2受体拮抗剂对大鼠纹体中DOPAC...

Dopamine metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) was detected by differential normal pulse voltammetry (DNPV) in chloral hydrate anesthetized rat striatum. The effects of some dopamine D-1 and D-2 antagonists on DOPAC concentration were observed in order to demonstrate the role of presynaptic D-1 and D-2 receptors in the feedback of dopamine synthesis and metabolism. The results showed that the mixed antagonists haloperidol (0.5 mg/kg s.c.) and chlorpromazine (5 mg/kg s.c.), the selective D-1 antagonist SCH23390 (1 mg/kg s.c.), and the selective D-2 antagonist sulpiride (5 mg/kg s.c.) all increased DOPAC concentration significantly. The percentages of increase were 140%, 102%, 26% and 40% in comparison with basal DOPAC concentration (14 +/- 2 mumol/L), respectively. These results indicate that not only presynaptic D-2 receptor regulates DA synthesis and metabolism, but that the presynaptic D-1 receptor also takes part in this feedback mechanism.     

D1 Receptor-Mediated Endogenous tPA Upregulation Contributes to Acute Blood Brain Barrier Damage

Background: Disruption of the blood brain barrier (BBB) integrity at the early stage of ischemia is becoming a critical target to reduce hemorrhage transformation (HT) because of the potential to predict HT. However, the mechanism underlying early BBB damage is not very clear. It was reported that after acute ischemia, there was a significant increase of dopamine release in striatum where we have reported BBB damage as well as upregulation of HIF-1α after 2-h ischemia. Objective: In current study, we aimed to investigate the role of dopamine signal pathway in BBB damage after acute ischemia using in vivo rat middle cerebral artery occlusion (MCAO) model. Results: Our data showed that there was an increase of endogenous tissue plasminogen activator (tPA) in BBB damage area and intra-striatum infusion of tPA inhibitor neuroserpin, significantly alleviated ischemia-induced BBB damage. In addition, intra-striatum infusion of D1 antagonist SCH23390 significantly decreased ischemia-induced upregulation of endogenous tPA, accompanied by decrease of BBB damage and occludin degradation. More important, inhibition of HIF-1 with inhibitor YC-1 significantly decreased acute ischemia-induced endogenous tPA upregulation and BBB damage. Conclusion: Taken together, our data demonstrate that acute ischemia disrupted BBB through activation of endogenous tPA via HIF-1 upreguation-induced dopamine increase, thus representing a new therapeutic target for protecting BBB, and may help alleviate HT following thrombolysis after ischemia stroke.