葛根素对帕金森病保护作用的实验研究

目的 :探讨葛根素能否对雌激素水平降低的帕金森小鼠提供保护作用 ,为开发具有雌激素替代作用的药物预防和治疗 PD提供新的线索。方法 :采用免疫组化染色 (ABC法 )计数酪氨酸羟化酶 (TH)的阳性细胞数 ,TUNEL法观察每视野下凋亡细胞数目 ,比较各组的差异。结果 :正常雌性小鼠 MPTP造模组 (A组 )比去势后 MPTP造模组 (B组 )中给予生理盐水组 (B1组 )、 MPTP造模后去势组 (C组 )给予生理盐水组 (C1组 )的 TH阳性细胞数目明显增多 ,凋亡细胞数目减少 (P<0 .0 5 ) ;B组中给雌激素组 (B2组 )与给葛根素组 (B3组 )相比无显著性差异 (P>0 .0 5 ) ;B2组、 B3组与 B1组相比有显著性差异 (P<0 .0 5 ) ;B1组与 C1组相比无显著性差异 (P>0 .0 5 ) ;在 C组中 ,给雌激素组 (C2组 )和给葛根素组 (C3组 )比对照组 (C1组 ) TH阳性细胞数目增多 ,凋亡细胞数目减少 (P<0 .0 5 ) ,C2组与 C3组相比无显著性差异 (P>0 .0 5 )。结论 :葛根素对帕金森病具有保护作用 ,并与凋亡有关     

不同剂量左旋多巴对帕金森病大鼠行为学影响实验研究

目的　观察不同剂量左旋多巴 (L - dopa)治疗帕金森病 (PD)的效果 ,探讨 L - dopa治疗帕金森病的合理方法。方法　通过 6- OHDA脑立体定向注射术建立大鼠 PD模型 ,采用行为学、TUNEL、原位杂交的方法观察左旋多巴小、中、大 3种不同剂量 [10 mg/(kg· d)、5 0 mg/(kg· d)、10 0 mg/(kg· d) ]、不同的作用时间 (1d、3 d、5 d、7d)对 PD大鼠黑质细胞的毒性作用 ,并观察治疗后 7天各项指标的变化。结果　与对照组比较 ,小剂量组大鼠旋转圈数随 L- dopa使用计量和时间增加而减少 ,中、大剂量组相反 ,旋转启动时间随 L- dopa使用剂量和时间增加而加速 ,最高转速及持续时间则相反。结论　我们应尽可能小剂量、间隔使用 L - dopa治疗 PD。     

不同剂量左旋多巴对正常大鼠黑质细胞作用的研究

目的　研究不同剂量左旋多巴对正常大鼠黑质细胞作用 ,探讨左旋多巴合理治疗帕金森病的方法。方法　采用TUNEL、Nissl染色 ,电镜观察手段 ,观察小、中、大 3种不同剂量 [10mg/(kg·d)、5 0mg/(kg·d)、2 0 0mg/(kg·d) ]左旋多巴 ,不同作用时间 (1天、7天、14天 )对正常大鼠黑质细胞作用。结果　对照组 ,小、中剂量左旋多巴给药组黑质细胞凋亡数 ,黑质细胞超微结构 ,尼氏细胞数无显著差异 ;大剂量给药组大鼠黑质细胞数随着左旋多巴使用时间、剂量的增加而增加 ,尼氏细胞则随左旋多巴使用时间、剂量增加而减少。结论　大剂量左旋多巴对正常大鼠黑质细胞有神经毒性作用 ,临床用左旋多巴宜尽可能小剂量     

雌激素受体α基因多态性与帕金森病的相关性

流行病学调查及动物实验提示，雌激素对帕金森病（Parkinson disease，PD）有保护作用，受体依赖途径是雌激素作用的重要环节，雌激素受体（estrogen receptor，ER）的基因多态性可能影响了PD的发病。我们拟探讨中国人群ERα的基因多态性与PD及PD痴呆（PD with dementia，PDD）的关系。     

植物雌激素对去势雌性大鼠缺血性脑损伤的神经保护作用

目的对比研究植物雌激素和动物雌激素对去势雌性SD大鼠永久性局灶性脑缺血组织的神经保护作用。方法采用线栓法建立右侧永久性大脑中动脉阻断模型。缺血24h后立即断头取脑,冠状切片,HE染色后于光镜下观察缺血侧大脑皮层的病理变化,应用免疫组化法检测不同实验组大鼠缺血侧Cfos表达情况,以DNA缺口末端标记法原位检测细胞凋亡,通过TTC染色比较脑梗死体积百分比。结果动物雌激素(17β雌二醇)组及植物雌激素(葛根素)组与生理盐水组相比,光镜下正常神经细胞密度显著增加,缺血侧C-fos表达阳性的细胞数明显减少,凋亡细胞数显著减少,脑梗死体积百分比也明显缩小(P<0.05)。两种雌激素相比,上述4项指标差异均无显著性(P>0.05)。结论两种雌激素对缺血性脑损伤均有保护作用,可减弱大脑缺血梗死灶边缘区的Cfos表达,从而延缓神经细胞凋亡。这可能是雌激素脑保护作用的机制之一。     

氧自由基在帕金森病发病中作用的实验研究

目的 观察氧自由基在帕金森病发病中的作用。方法 大鼠分为3组：帕金森病组、假手术组、对照组。帕金森病大鼠模型采用6羟基多巴胺(6—OHDA)制作。采用生化、电镜方法观察铁离子、氧自由基浓度及黑质超微结构变化。结果 帕金森病组大鼠铁离子、氧自由基浓度及黑质损伤程度均高于对照组和假手术组。结论 氧自由基在帕金森病发病中起着重要作用。     

雌激素对MPP+诱导的PC12细胞损伤的防护作用

目的　探讨在离体的细胞培养物中 ,雌激素 (estrogen ,E)对 1 甲基 4 苯基吡啶离子(MPP+ )诱导的帕金森模型是否具有保护作用。方法　以PC12细胞为多巴胺能神经元的细胞模型 ,将MPP+ 或E加入培养的PC12细胞 ,四甲基偶氮唑盐 (MTT)法检测细胞活性及代谢状态 ,SABC法检测酪氨酸羟化酶 (TH)的含量 ,流式细胞术检测细胞凋亡率 ,比较各组的差异。结果　随MPP+ 浓度增加 ,细胞活性下降 ,MPP+ 浓度为 2 5 0 μmol/L时 ,吸光度A570 值为 0 30± 0 0 7,符合帕金森病细胞模型 ;随E浓度增加 ,细胞活性升高 ,呈量效依赖关系 ;当雌激素浓度 >10nmol/L ,细胞活性无明显增加。单纯用E组A570 值为 0 6 1± 0 17,比空白对照组 (0 4 9± 0 11)、MPP+ 组 (0 30± 0 0 7)、E +MPP+ 组 (0 5 6± 0 16 )细胞活性高 (P <0 0 5 ) ;TH阳性细胞平均吸光度E组 (0 4 6± 0 0 6 )比空白对照组 (0 2 2± 0 0 7)、MPP+ 组 (0 10± 0 0 3)、E +MPP+ 组 (0 2 4± 0 0 4 )高 (P <0 0 5 ) ;凋亡率E组(11 5 % )比对照组 (31 3% )、MPP+ 组 (6 3 5 % )、E +MPP+ 组 (33 6 % )低 (P <0 0 5 ) ,每组细胞数为 2× 10 5个 /ml。E +MPP+ 组比MPP+ 组细胞活性高 ,TH含量高 ,凋亡率低 (P <0 0 5 )。结论　雌激素对PC12细胞可     

雌激素对去卵巢大鼠缺血再灌注脑损伤的保护作用

目的探讨雌激素对去卵巢大鼠缺血再灌注脑损伤的保护作用。方法将大鼠切除双侧卵巢后30d给予肌肉注射苯甲酸雌二醇100μg/(kg·d)连续14d,然后制作大鼠局灶性脑缺血再灌注模型;制模12h后取脑组织行免疫组化染色,检测细胞间黏附分子(CD54)、肿瘤坏死因子α(TNF-α)的表达;TUNEL法检测脑组织凋亡细胞数;电镜观察脑细胞膜超微结构的变化。结果与缺血再灌注组及去卵巢组比较,雌激素组脑组织CD54、TNF-α表达明显降低,凋亡细胞数明显减少(均P<0.05);脑细胞膜结构非特异性损伤减轻。结论雌激素通过减少缺血再灌注大鼠脑组织炎性细胞因子表达、减轻炎症反应、降低脑组织细胞凋亡而发挥脑保护作用。     

雌激素对大鼠脑缺血-再灌注损伤的保护作用的研究

目的　研究外源性雌激素替代对大鼠局灶性脑缺血 再灌注损伤的神经保护作用。方法　将12 0只大鼠分为对照组、双侧卵巢切除 (OVX)组、OVX +雌二醇 (E2 )组 (E2 2 0 0 μg/kg ,皮下注射每周 1次 ,连续 4周 )。 4周后用线栓法建立脑缺血再灌注模型 ,在缺血再灌注不同时间观察大鼠脑梗死体积、病理改变、凋亡细胞数及检测血浆雌激素水平。结果　缺血再灌注不同时间均以OVX +E2 组脑梗死体积最小 ,OVX组最大 ,两组比较有显著差异 (均P <0 0 1)。凋亡细胞数OVX组明显多于OVX +E2 组 (P <0 0 5 )。结论　雌激素对缺血 再灌注后脑损伤具有明显的保护作用     

美满霉素对帕金森病细胞凋亡模型保护作用的研究

目的探讨美满霉素(MC)对1甲基4苯基吡啶离子(MPP+)诱导的帕金森病细胞凋亡模型保护作用的机制。方法将不同浓度(10、50、250、500μmol/L)MPP+加入培养的PC12细胞中,选择最适当浓度的MPP+建立多巴胺神经元凋亡模型(MPP+组);通过四甲基偶氮唑盐法(MTT法)检测经不同浓度(0、10、50、100、200μmol/L)MC预处理后多巴胺神经元凋亡模型的活性,以此筛选出具有最佳保护作用MC浓度建立MC+MPP+组;用电泳法、流式细胞术和逆转录聚合酶链式反应分别检测MPP+组、MC+MPP+组细胞的凋亡率,以及此2组细胞caspase3mRNA的表达,并与空白对照组比较。结果(1)在MPP+为10μmol/L时可见细胞凋亡最典型的梯状DNA条带,以此浓度建立多巴胺神经元凋亡模型(MPP+组);用100μmol/LMC预处理的MPP+组细胞活性最高(P<0.05),以此作为此后实验用MC浓度。(2)MC+MPP+组细胞凋亡率及caspase3mRNA的灰度比值分别为(22.83±2.10)%及68.08±1.14,极显著低于MPP+组的(45.89±2.28)%及86.50±1.43(均P<0.01),但仍明显高于空白对照组的(11.05±1.02)%及53.75±1.23(均P<0.05)。结论MC可能通过下调caspase3mRNA表达保护MPP+诱导的PC12细胞凋亡。     

帕金森病与QTc间期的关系

目的：观察帕金森病（PD）与QTc间期的关系。方法：分析了37例PD患者及37例健康对照组的QTc间期,并比较PD病病情与QTc间期的关系,结果：PD组QTc间期明显超过对照组（P＝0．001）,PD病情的轻重可影响QTc间期。结论：对QTc间期的监测有助于发现PD患者自主神经功能障碍,同时可作了为解病情轻重的一个参考指标。     

One-Year Open-Label Study of Entacapone in Patients with Advanced Parkinson Disease

Background and purpose

A carboxy-O-methyl transferase inhibitor entacapone has been introduced as an adjuvant drug for Parkinson disease (PD) patients. Although clinical trials reported beneficial role of entacapone, a long-term trial over 3 years failed to show significant effect. The goals of this study were to evaluate the clinical benefit and the efficacy of entacapone in an open clinical practice.

Methods

After the completion of a double-blind placebo-controlled entacapone study, 149 patients from 4 centers were included. Antiparkinsonian medications were optimized by the judgment of the neurologists in charge. The clinical global impression (CGI) scale was obtained at 6 months and 1 year after the initiation of entacapone treatment.

Results

Of the 149 patients, 117 patients chose to try entacapone in an open-label fashion. Sixty-nine (59%) patients completed the 1-year trial. Twenty-nine patients discontinued entacpaone before 6 months, and 19 between 6 months and 1 year during trial. Twelve patients out of 48 patients discontinued entacapone because of its poor efficacy. The CGI scale was 3.9 (±1.5) at the beginning of the trial, 4.3 (±1.1) at 6 month, and 3.8 (±1.3) at 1 year, respectively. The CGI scale of those who discontinued between 6 month and 1 year was 3.4 (±1.7), which was worse, but insignificantly, than that of the continuer.

Conclusions

The dropout at 1 year of our study was very high at 41%. Even though entacapone is indicated for advanced PD patients with motor fluctuation, the fluctuators commonly have dyskinesia and mental symptoms, which can become more troublesome with entacapone. In the patients with advanced PD, the clinical efficacy and side effects should be carefully considered in a long-term use of entacapone.     

铁在大鼠黑质多巴胺能神经元损伤中作用研究

目的：观察铁在多巴胺能神经元损伤过程中的作用，并探讨铁在帕金森病Parkinson disease，PD)起病阶段的作用。方法：通过PD大鼠模型，采用生化、TUNEL方法观察6-羟基多巴胺(6-OHDA)脑立体定向注射1、7、14、21d PD黑质铁浓度、自由基以及黑质细胞凋亡数变化。结果：1～21d PD大鼠黑质铁浓度、自由基及凋亡细胞数随时间增加而增加，并显著高于对照组。结论：铁在PD发病中具有重要作用。     

Genetics of Parkinson Disease

Summary: Parkinson disease (PD) is the second most common neurodegenerative disorder. Recent studies have consistently demonstrated that in some families, disease is attributable to a mutation in a single gene. To date, genetic analyses have detected linkage to six chromosomal regions and have identified three causative genes: PARK1 (alpha-synuclein), PARK2 (parkin), and PARK7 (DJ-1). In addition, mutations in several other genes have been implicated in familial PD. Identification of the mutations in these genes has led to the recognition that the ubiquitin-proteasome system is an important pathway that may be disrupted in PD. Studies are ongoing to identify additional genes that may contribute to PD susceptibility, particularly in late-onset families without a clear pattern of disease inheritance. With the identification of mutations in particular genes and the likely role of additional genes that are important in PD risk-susceptibility, appropriate protocols must be developed so that accurate and informative genetic counseling can be offered to families in which one or more members has PD. Further diagnostic testing should be delayed until more is learned about the frequency, penetrance, and risk assessment of certain gene mutations. Important lessons can be learned from the implementation of counseling protocols for other neurodegenerative disorders, such as Huntington disease and Alzheimer disease.     

被动吸烟对帕金森病大鼠旋转行为和纹状体多巴胺含量的影响

目的：观察被动吸烟对帕金森病（PD）大鼠的影响,以验证流行病学研究的结论,为PD研制提供一新的线索。方法：用6－羟基多巴胺（6－OHDA）立体定向注入大鼠一侧黑质致密部和中脑被盖腹侧区建立侧侧PD模型,观察术前4周开始始予的被动吸烟（持续6周）和术后2周对成功模型给予的被动吸烟（持续2周）对阿朴吗啡诱发的旋转行为及纹状体DA含量的影响。结果：术前4周开始被动吸烟的大鼠旋转行为有减少趋势,受损侧纹状体DA含量较对照组升高,术后2周,成功模型给予的被动吸烟对PD大鼠的旋转行为及纹状体DA含量均无影响。结论;被动吸烟可减轻6－OHDA对黑质DNA能神经元的损伤。     

Protective mechanism of Wnt4 gene on Parkinson’s disease (PD) transgenic Drosophila

Objective: The main pathological change of Parkinson’s disease (PD) is progressive degeneration and necrosis of dopaminergic neurons in the midbrain, forming a Lewy body in many of the remaining neurons. Studies have found that in transgenic Drosophila, mutations in the PTEN-inducible kinase 1 (PINK1) gene may cause indirect flight muscle defects in Drosophila, and mitochondrial structural dysfunction as well.

Methods: In this study, Wnt4 gene overexpression and knockdown were performed in PINK1 mutant PD transgenic Drosophila, and the protective effect of Wnt4 gene on PD transgenic Drosophila and its possible mechanism were explored. The Wnt4 gene was screened in the previous experiment; And by using the PD transgenic Drosophila model of the MHC-Gal4/UAS system, the PINK1 gene could be specifically activated in the Drosophila muscle tissue.

Results: In PINK1 mutation transgenic fruit flies, the Wnt4 gene to study its implication on PD transgenic fruit flies’ wing normality and flight ability. We found that overexpression of Wnt4 gene significantly reduced abnormality rate of PD transgenic Drosophila and improved its flight ability, and then, increased ATP concentration, enhanced mitochondrial membrane potential and normalized mitochondrial morphology were found. All of these findings suggested Wnt4 gene may have a protective effect on PD transgenic fruit flies. Furthermore, in Wnt4 gene overexpression PD transgenic Drosophila, down-regulation autophagy and apoptosis-related proteins Ref(2)P, Pro-Caspase3, and up-regulation of Beclin1, Atg8a, Bcl2 protein were confirmed by Western Blotting.

Conclusion: The results imply that the restoring of mitochondrial function though Wnt4 gene overexpression in the PINK1 mutant transgenic Drosophila may be related to autophagy and/or apoptosis.     

银杏叶提取物治疗帕金森病大鼠的研究

目的　观察银杏叶提取物 (EGb)治疗帕金森病 (PD)的效果 ,论证EGb治疗PD的可能性。方法　通过PD大鼠模型 ,采用免疫组织化学、HE染色、电镜及旋转行为检查的方法 ,观察模型成功后EGb(EGb组 )治疗 1天、7天、14天大鼠旋转行为及黑质病理变化 ,并与对照组 (PD组 )比较。结果　EGb组酪氨酸羟化酶染色显著高于PD组 (P <0 0 5 ) ,旋转圈数、黑质bFGF蛋白表达均较PD组减少 (均P <0 0 5 ) ,EGb组黑质超微结构及HE染色结构损伤均减轻。结论　银杏叶提取物可能是治疗PD的有效药物