环加氧酶-2对帕金森病模型小鼠黑质多巴胺能神经元的影响

目的　观察COX 2对帕金森病小鼠黑质多巴胺能神经元的影响。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。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 (Ty rosineHydroxylase ,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很可能与帕金森病时黑质多巴胺能神经元的损伤有关     

p38MAPK抑制剂对MPTP模型小鼠黑质多巴胺能神经元的保护作用

目的:研究SB239063在1-甲基-4-苯基-1,2,3,6四氢吡啶(MPTP)所致帕金森病(PD)模型小鼠中抑制p38丝裂原激活蛋白激酶(mitogen-activated protein kinase,MAPK)活化对多巴胺(DA)能神经元的保护作用。方法:雄性C57BL/6N小鼠随机分为MPTP(30 mg/kg)模型组;SB239063(5 mg/kg)抑制剂组;SB239063(15 mg/kg)抑制剂组;SB239063(25 mg/kg)抑制剂组。抑制剂组于每次注射MPTP前3 h腹腔注射SB239063;对照组注射与模型组和抑制剂组等量生理盐水和DMSO。采用免疫组织化学和免疫蛋白印迹法观察各组小鼠黑质酪氨酸羟化酶(TH)和磷酸化p38蛋白激酶(p-p38 protein kinase,p-p38MAPK)之间表达变化的关系。结果:模型组小鼠黒质区p38MAPK显著活化,同时伴有TH阳性神经元明显丢失;SB239063 15 mg/kg与25 mg/kg组均可明显减少TH神经元丢失,而5 mg/kg组无显著影响;免疫荧光双标记结果显示p38MAPK与TH阳性神经元存在共表达。结论:p38MAPK对PD模型小鼠中脑黑质多巴胺能神经元丢失可能有重要调控作用,SB239063对多巴胺神经元具有一定的神经保护作用。     

帕金森病小鼠黑质多巴胺能神经元凋亡诱导因子核移位的研究

目的探讨帕金森病(PD)小鼠模型黑质凋亡诱导因子(AIF)的核移位情况及其与多巴胺(DA)能神经元损伤之间的关系。方法采用1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)制备PD小鼠模型,2h、24h、72h后取中脑组织进行酪氨酸羟化酶免疫染色观察黑质DA能神经元的损害情况,AIF免疫组化染色和Western blot方法检测AIF的核移位情况。结果与对照组比较,PD组小鼠黑质DA能神经元数量随时间呈递减趋势,并出现AIF核移位,2h时达高峰,之后随时间呈下降趋势。结论AIF核移位是PD小鼠黑质DA能神经元损伤的早期指标,在PD的发病过程中发挥重要作用。     

JNK信号通路对亚急性帕金森病MPTP模型小鼠黑质iNOS表达的影响

目的:研究JNK在1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)所致小鼠帕金森病(PD)模型中对诱导型一氧化氮合酶(iNOS)表达的调控作用,探讨PD黑质多巴胺(DA)能神经元变性失活的可能机制.方法:采用神经毒素MPTP制备亚急性PD小鼠模型,健康雄性C57BL/6N小鼠随机分为模型组、JNK抑制剂组和对照组,采用行为学观察、免疫组织化学和免疫印迹法,观察模型小鼠行为学变化及中脑黑质酪氨酸羟化酶(TH)、 iNOS和磷酸化c-Jun(p-c-Jun)的表达变化;观察给予JNK通路特异性抑制剂SP600125对上述变化的影响.结果:与对照组相比,模型小鼠出现典型PD样症状.中脑黑质区TH阳性神经元下降约65%,中脑黑质TH表达水平显著降低约75%;黑质区iNOS和p-c-Jun阳性细胞明显增加,且p-c-Jun特异性表达于细胞核,中脑黑质iNOS与p-c-Jun表达水平明显升高.经SP600125处理后,模型小鼠PD样症状减轻,与对照组比较,TH阳性细胞数和TH表达水平仅下降约15%和25%,与模型组比较,黑质区iNOS与p-c-Jun阳性细胞减少,且p-c-Jun仅表达于胞质,中脑黑质iNOS与p-c-Jun表达水平明显下降.结论:JNK通路在MPTP诱导的亚急性PD小鼠黑质iNOS表达中可能起重要的调控作用;JNK通路特异性抑制剂SP600125可能对帕金森病小鼠具有一定的神经保护作用.     

慢性帕金森病模型小鼠的行为学表现和多巴胺能神经细胞及转运体的数量变化

目的:研究慢性帕金森病模型小鼠行为学、黑质致密部多巴胺神经细胞和纹状体多巴胺转运体(DAT)的变化。方法:利用MPTP和二丙苯磺胺联合注射小鼠5周(每周2次,共10次)制作帕金森病小鼠慢性模型,通过转棒法检测小鼠行为学表现,免疫组织化学方法测定黑质内多巴胺能神经细胞的变化,放射自显影技术测定纹状体内DAT的密度。结果:帕金森病小鼠在转棒试验中行为发生明显改变,模型组行为学得分明显低于对照组(895±238 vs 1202±252,P0.001),酪氨酸羟化酶(TH)阳性细胞数也明显低于对照组(10.4±6.5 vs22.4±8.0,P0.001)。模型组动物纹状体DAT放射自显影灰度值明显低于对照组(0.2123±0.0316 vs 0.3034±0.0588,P0.001)。线性回归表明,模型组小鼠黑质致密部TH阳性细胞数及纹状体内DAT密度与对应行为学得分呈明显正相关(r=0.6354,P0.01)。结论:MPTP和二丙苯磺胺所致的帕金森病小鼠,在行为学、纹状体DAT密度和黑质致密部多巴胺神经细胞量上均有明显改变,提示这种小鼠模型可用于研究帕金森病的发病过程和药物治疗。     

虫草素对MPTP诱导的帕金森病小鼠运动与认知能力的影响

目的:探索虫草素(cordycepin)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)诱导的帕金森病小鼠运动与认知能力的影响。方法:将C57BL/6小鼠每天腹腔注射MPTP (30 mg/kg),连续8 d,以建立帕金森病小鼠模型。HE染色检测虫草素对MPTP诱导小鼠黑质致密部(sub-stantia nigra pars compacta,SNpc)细胞数目的影响;Western blot检测黑质(substantia nigra,SN)内酪氨酸羟化酶(tyrosine hydroxylase,TH)表达量的变化;采用旷场实验(open-filed test,OFT)、自发交替行为(spontaneous alterna-tion behavior,SAB)及水迷宫实验(water maze test,WMT)分别检测虫草素对帕金森病小鼠自发活动、情绪变化和认知行为的影响。结果:HE染色结果表明,虫草素显著抑制由MPTP引起的小鼠SNpc细胞数目减少(P 0.05);Western blot结果表明MPTP显著降低SN内TH的表达量(P 0.05),而虫草素可以逆转这种变化(P 0.05);虫草素可提高小鼠在OFT中的平均速度(P 0.05),并增加小鼠在SAB中的总进臂次数,提高正确率(P 0.05),但对WMT潜伏期并没有明显的影响。结论:虫草素能够减轻MPTP诱导的帕金森病小鼠早期的运动障碍和探索能力降低,但对其记忆障碍并没有明显的影响。     

银杏叶提取物对帕金森病模型小鼠多巴胺能神经元保护作用研究

目的研究银杏叶提取物（GBE）对帕金森病模型小鼠黑质（SN）多巴胺（DA）能神经元的保护作用。方法36只C5,Bk小鼠随机3组,每组12只。其中,帕金森病（PD）模型组的小鼠采用1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）腹腔注射（30mg／kg×5d）诱导PD;GBE预处理组的小鼠于注射MPTP前2h腹腔注射GBE（60mg／kg×8d）;正常对照组的小鼠只注射等体积生理氯化钠溶液。应用免疫组织化学染色观察小鼠黑质致密带（SNzc）酪氨酸羟化酶（TH）免疫反应阳性细胞数量的变化,用高效液相色谱法（HPLC-ECD）检测小鼠纹状体（Str）内DA及其代谢物二羟基苯乙酸（DOPAC）和高香草酸（HVA）含量。结果PD模型组小鼠SN内酪氨酸羟化酶（TH）阳性细胞数量、Str内DA及其代谢产物DOPAC和HVA的含量明显减少（P〈0．01）,GBE预处理组小鼠SN内TH阳性细胞数量、Str内DA及其代谢产物DOPAC和HVA的含量明显增多（P〈0.05）,但仍低于正常对照组（P〈0．01）。结论GBE对MPTP致帕金森病小鼠SNDA能神经元具有明显的保护作用。     

MPTP诱导的帕金森病模型小鼠味觉偏好变化

目的:探讨1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)对帕金森病(PD)模型小鼠味觉的影响。方法:将雄性C57BL/6J小鼠分成对照组(control)和MPTP组,通过转棒、爬杆、嗅觉和悬尾实验检测小鼠行为,组织免疫荧光染色检测小鼠黑质致密部(SNpc)多巴胺(DA)能神经元。采用双瓶偏好实验检测小鼠对蔗糖、糖精、氯化钠及奎宁溶液的偏好率。利用real time RT-PCR检测小鼠舌上皮TAS1R1、TAS1R2和TAS1R3的mRNA表达的变化。结果:与control组相比,MPTP组小鼠转棒时间显著性降低,爬杆时间、找到隐藏食物的时间以及悬尾实验中静止时间均显著增加,免疫荧光结果显示MPTP处理可显著减少SNpc的DA能神经元数量。通过双瓶偏好实验发现MPTP可诱导小鼠对蔗糖、糖精、氯化钠的偏好率增加;但对苦味剂奎宁的偏好率没有影响。qRT-PCR分析发现MPTP抑制了小鼠舌上皮味觉受体TAS1R1和TAS1R2 mRNA的表达,而对TAS1R3 mRNA的表达没有影响。结论:MPTP诱导的PD模型小鼠对蔗糖、糖精、氯化钠的饮用偏好率增加,而对奎宁的偏好率没有影响,...     

磷酸化P38 MAPK对帕金森病MPTP模型小鼠黑质caspase-3表达的影响

目的研究磷酸化P38 MAPK在1-甲基-4-苯基-1,2,3,6四氢吡啶(MPTP)所致帕金森病(PD)模型小鼠中对黑质半胱氨酸蛋白酶-3(caspase-3)的调控作用。方法将小鼠随机分为MPTP模型组,腹腔注射MPTP(30mg/kg,生理盐水溶);抑制剂组,在注射MPTP前1h腹腔注射SB203580(10mg/kg,溶于5mg/mLDMSO)。均1次/d,连续5d;对照组,注射与模型组和抑制剂组等量生理盐水和DMSO。观察行为学、免疫组织化学和免疫蛋白印迹法观察黑质酪氨酸羟化酶(TH)、caspase-3和磷酸化P38 MAPK(p-P38MAPK)的表达。结果与对照组相比,模型小鼠出现典型的PD症状,TH阳性神经元和蛋白水平分别下降约60%和65%(P<0.01),p-P38 MAPK、caspase-3阳性细胞及蛋白水平显著增加(P<0.01);经P38 MAPK抑制剂SB203580处理后,上述变化均显著减轻(P<0.01)。结论磷酸化P38 MAPK在MPTP诱导的PD小鼠黑质caspase-3表达中可能有重要调控作用,SB203580对PD小鼠具有一定的神经保护作用。     

Nrf3基因在帕金森病猴模型黑质中表达的研究

本研究目的在于采用 m RNA差异显示技术及改进的硝酸银核酸染色技术 ,寻找 1-甲基 -4 -苯基 -1,2 ,3,6 -四氢吡啶诱导侧与对照侧中脑黑质细胞之间的差异表达基因。用 1-甲基 -4 -苯基 -1,2 ,3,6 -四氢吡啶诱发单侧帕金森病猴模型后 ,将两侧黑质m RNA用 3’端三个锚定引物与 30种随机引物进行逆转录 -聚合酶链反应 ,经聚丙烯酰胺凝胶电泳及硝酸银染色后显示扩增产物条带。结果 :对获得的差异表达基因片段的序列分析、同源性比较及功能分析证明 ,差异基因中的一个 (14 9bp)为 Nrf3基因片段。此结果提示 :Nrf3可能参与小胶质细胞的生长 ,促进对变性神经元的吞噬作用 ,可为从分子水平了解人类帕金森病的发生机理提供依据     

Genistein对Parkinson病模型小鼠黑质纹状体通路的保护作用

为了研究大豆异黄酮活性成分genistein对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)制备的去卵巢(OVX)Parkinson病(PD)模型小鼠黑质纹状体通路的保护作用,我们将OVX PD小鼠随机分为对照组、MPTP组、genistein预处理组和雌激素预处理组,采用免疫组织化学染色结合逆转录-聚合酶链式反应法(RT-PCR)检测黑质致密带(SNpc)神经元酪氨酸羟化酶(TH)的表达情况,采用高效液相色谱法(HPLC-ECD)检测小鼠纹状体(Str)多巴胺(DA)及其代谢物二羟基苯乙酸(DOPAC)和高香草酸(HVA)的含量。结果显示:Genistein及雌激素用药组SN内TH阳性神经元数量及THmRNA的表达水平较MPTP组显著升高(P<0.01)。MPTP组Str内DA及其代谢产物DOPAC和HVA的含量较对照组明显降低(P<0.01)。genistein及雌激素用药组Str内DA、DOPAC及HVA含量较MPTP组显著升高(P<0.01)。上述结果提示genistein对MPTP制备的PD模型小鼠黑质DA能神经元有明显的保护作用。     

小白菊内酯对帕金森病小鼠模型黑质多巴胺能神经元的保护作用

目的:探讨小白菊内酯在帕金森病小鼠模型中抑制核因子NF-κB(nuclear factor kappa B)信号通路对多巴胺(DA)能神经元的保护作用。方法:将C57BL/6N小鼠随机分为模型组、干预组和对照组。通过行为学、免疫组织化学和免疫蛋白印记法观察各组小鼠中脑黑质区酪氨酸羟化酶(TH)、核因子NF-κB、环氧合酶-2(COX-2)和半胱氨酸天冬氨酸蛋白酶-3(caspase-3)的表达变化情况。结果:与对照组相比,模型组小鼠出现典型PD症状,TH阳性神经元明显丢失、蛋白水平下降,NF-κB、COX-2和caspase-3阳性细胞大量表达,蛋白水平显著升高(P0.01)。抑制剂组小鼠上述变化明显减轻(P0.01)。结论:在MPTP所致PD模型小鼠中,小白菊内酯通过抑制核因子NF-κB信号通路减少凋亡蛋白caspase-3表达,从而发挥神经保护作用。     

尼莫地平对帕金森病模型小鼠黑质多巴胺神经元的保护作用

目的:研究尼莫地平在1-甲基-4-苯基-1,2,3,6四氢吡啶(MPTP)所致帕金森病(PD)模型小鼠中对黑质多巴胺能神经元的保护作用。方法:雄性健康C57BL/6N小鼠随机分为对照组,腹腔注射生理盐水;模型组,腹腔注射MPTP(25 mg/kg);尼莫地平治疗组,每次注射MPTP前3 h灌胃给予尼莫地平(15 mg/kg)。观察各组小鼠行为学变化,免疫组织化学和免疫蛋白印迹法观察中脑黑质酪氨酸羟化酶(TH)、前列腺素E2(PGE2)和肿瘤坏死因子α(TNF-α)的表达变化。结果:与对照组小鼠比较,MPTP模型组小鼠出现典型的PD症状,中脑黑质TH阳性神经元大量丢失,PGE2和TNF-α阳性细胞显著增多,蛋白水平明显升高;经尼莫地平治疗后,上述症状得到明显改善。结论:在MPTP所致PD小鼠模型中,尼莫地平通过抑制炎症因子PGE2和TNF-α的表达从而对多巴胺能神经元具有一定的保护作用。     

罗格列酮对帕金森病小鼠黑质内多巴胺能神经元的保护作用

目的:研究罗格列酮在1-甲基-4-苯基-1,2,3,6四氢吡啶(MPTP)所致帕金森病(PD)模型小鼠中对多巴胺能神经元的保护作用.方法:采用MPTP制备PD小鼠模型,观察各组小鼠行为学变化,免疫组织化学和免疫印迹观察中脑黑质酪氨酸羟化酶(TH)、环氧合酶(COX-2)和肿瘤坏死因子-α(TNF-α)的表达变化,及给予罗格列酮后对上述变化的影响.结果:模型组小鼠出现震颤、步态迟缓等PD样症状,黑质区TH阳性神经元缺失,炎症因子COX-2和TNF-α阳性细胞明显增多,蛋白水平亦升高;经罗格列酮治疗后,上述症状得到改善.结论:罗格列酮对模型小鼠多巴胺能神经元保护作用可能抑制炎症因子COX-2和TNF-α表达,从而减轻炎症反应.     

Effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor on respiratory burst activity of neutrophiis in patients with myelodysplastic syndromes

The superoxide (O₂-releasing capacity in response lo N-fomiyl-methionyl-leucyl-phenylalanine (FMLP)and the priming effects ofrecombinant human granuloeyte colony-stimulating factor (rhG-CSF) and granulocyte-macrophage colony-stimulating faclor (rhGM-CSF) on FMLP-induced O₂ release were investigated in neutrophils from 14 patients with myelodysplastic syndromes (MDS). The O₂ -releasing capacity in MDS neutrophils varied from patient to patient. As compared with normal neutJ-ophils. theO₂-releasing capacity in MDS neutrophils was increased in 9/14 patients, nonnal in three patients and decreased in two patients. There was no close relationship between the 02-reIeasing capacity and the peripheral blood neutrophil count or the plasma concentration of C-reactive protein. The priming of neutrophils by rhG-CSF was not observed in five patients, whereas rhGM-CSF primed neutrophils from all patients. The priming eflect of rhGM-CSF was consistently greater than that of rhG-CSF in each patient. The intravenous administration of rhG-CSF (300 μg/body) to two MDS patients showed an increase in the peripheral blood neutrophil count and enhancement of neutrophil O₂ release. These findings demonstrate that the neutrophil O₂-releasing capacity in MDS varies from patient to patient and is not always impaired, and that rhGM-CSF is able to prime neutrophils which never respond to rhG-CSF.     

丙戊酸钠对帕金森病模型小鼠多巴胺能神经元及脑源性神经营养因子表达的影响

帕金森病(Parkinson's disease,PD)是一种神经系统退行性疾病,主要表现为中脑黑质致密部多巴胺能神经元丧失,纹状体多巴胺含量下降,至今尚无有效的治疗手段.丙戊酸钠(Valproate, VPA)是临床上作为治疗双相精神障碍的主要药物.     

Effects of granulocyte colony-stimulating factor and hyaluronidase on the formation of blood system reactions

We studied the possibility of modification of hematotropic effects of granulocytic CSF and hyaluronidase. It was found that hyaluronidase in a dose of 20 U/mouse potentiates the specific effect of granulocytic CSF on granulocytopoiesis, while granulocytic CSF potentiates the stimulating effect of the enzyme on the erythroid stem. Functional activity of hemopoiesis precursors, secretion of humoral factors by adherent myelokaryocytes, and serum content of hemopoietins increased under these conditions. Hyaluronidase (100 U/mouse) against the background of treatment with granulocytic CSF leads to uncoupling of proliferation and differentiation of hemopoietic cells and abolishes the mutually activating hematotropic effect of preparations. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 145, No. 6, pp. 628–633, June, 2008     

Effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor on respiratory burst activity of neutrophils in patients with myelodysplastic syndromes.

The superoxide (O2-)-releasing capacity in response to N-formyl-methionyl-leucyl-phenylalanine (FMLP) and the priming effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and granulocyte-macrophage colony-stimulating factor (rhGM-CSF) on FMLP-induced O2- release were investigated in neutrophils from 14 patients with myelodysplastic syndromes (MDS). The O2(-)-releasing capacity in MDS neutrophils varied from patient to patient. As compared with normal neutrophils, the O2(-)-releasing capacity in MDS neutrophils was increased in 9/14 patients, normal in three patients and decreased in two patients. There was no close relationship between the O2(-)-releasing capacity and the peripheral blood neutrophil count or the plasma concentration of C-reactive protein. The priming of neutrophils by rhG-CSF was not observed in five patients, whereas rhGM-CSF primed neutrophils from all patients. The priming effect of rhGM-CSF was consistently greater than that of rhG-CSF in each patient. The intravenous administration of rhG-CSF (300 micrograms/body) to two MDS patients showed an increase in the peripheral blood neutrophil count and enhancement of neutrophil O2- release. These findings demonstrate that the neutrophil O2(-)-releasing capacity in MDS varies from patient to patient and is not always impaired, and that rhGM-CSF is able to prime neutrophils which never respond to rhG-CSF.     

Effects of recombinant human granulocyte colony-stimulating factor on neutropenia in patients with congenital agranulocytosis

Congenital agranulocytosis is a disorder characterized by severe neutropenia and a profound deficiency of identifiable neutrophil progenitors in bone marrow. In an attempt to stimulate neutrophil production and thereby reduce the morbidity and mortality associated with this disease, we administered recombinant human granulocyte colony-stimulating factor (rhG-CSF) in doses of 3 to 60 micrograms per kilogram of body weight per day to five patients with congenital agranulocytosis. In all five patients, an increase in the number of neutrophils was noted eight to nine days after the initiation of the effective dosage (the dose at which the neutrophil count reached 1000 cells per microliter or more and the bone marrow showed granulocyte maturation beyond the myelocyte stage). The absolute neutrophil counts rose from less than 100 to between 1300 and 9500 cells per microliter. Marrow aspirates obtained after 14 days at the effective dosage showed maturation to the mature neutrophil stage. The side effects that were observed were medullary pain, splenomegaly, and an elevation of levels of leukocyte alkaline phosphatase. All five patients have had sustained neutrophil counts of 1000 cells per microliter or more for 9 to 13 months while receiving subcutaneous maintenance therapy. Preexisting chronic infections have resolved clinically, and the number of new infectious episodes and the requirement for intravenous antibiotics have decreased. We conclude that treatment with rhG-CSF can lead to a large increase in the numbers of functional neutrophils in patients with congenital agranulocytosis.