蛋白酶体抑制剂诱导动物帕金森病模型的研究进展

帕金森病是一种好发于中老年人的神经变性疾病,其发病机制不明.目前,应用蛋白酶体抑制剂诱导的帕金森病动物模型,其行为学、影像学、病理学和生化特征都与帕金森病的表现极为相似,它的出现将会为帕金森病发病机制及治疗方法的探讨提供新途径.     

蛋白酶体抑制剂诱导帕金森病大鼠模型海马区HSP70表达

目的观察蛋白酶体抑制剂Lactacystin诱导帕金森病大鼠模型海马区HSP70表达。方法将Lactacystin 10μg（2μl）经立体定向仪注射到大鼠（Lactacystin组,n=36）左侧黑质致密部,对照组（n=36）以等体积生理盐水代替;分别在药物注射后24h、3d、5d、7d、9d、11d、14d、18d和21d提取海马标本,通过免疫组化和RT-PCR观察HSP70的表达。结果Lactacystin组大鼠海马区HSP70表达较对照组高,表现为注药24h后表达开始增高,5d达高峰,11d开始减少;且海马区表达分布呈CA3〉CA2〉CA1。结论Lactacystin诱导帕金森病大鼠海马区HSP70表达增高,且呈明显区域性和时间依从性;增加的热休克蛋白可能参与拮抗蛋白酶体抑制毒性而起到神经细胞保护作用。     

蛋白酶体抑制剂诱导多巴胺能神经元变性及诱导型一氧化氮合酶变化

目的探讨蛋白酶体（proteasome）功能下降在帕金森病（PD）发病机制中的作用，以及模型大鼠脑内黑质部位诱导型一氧化氮合酶（iNOS）是否参与蛋白酶体抑制剂Lactacystin诱导的多巴胺能神经元变性。方法将30只健康雄性SD大鼠分为5组（生理盐水对照组，1d组、3d组、1周组、3周组），每组6只。将蛋白酶体抑制剂Lactacystin立体定向注射至大鼠黑质部位，记录大鼠在不同时间点的行为学改变，并用免疫组化方法观察生理盐水对照组及不同时间点组（1d、3d，1周、3周）大鼠黑质区多巴胺能神经元变性及iNOS变化。结果Lactacystin注射1周后大鼠开始出现自发性活动减少，阿朴吗啡可诱导出旋转行为；3周后，30min旋转次数为258．90±11．56；实验3周组黑质部位TH阳性细胞减少。1d后iNOS阳性细胞明显增多，3d时达高峰，1周后开始下降，3周时基本消失。结论蛋白酶体功能下降可能是多巴胺能神经元变性的始动因素，而iNOS上调可能是多巴胺能神经元变性的重要过程。     

蛋白酶体抑制剂诱导大鼠运动行为改变的研究

目的 观察蛋白酶体抑制剂诱导大鼠黑质变性及大鼠运动行为改变,探讨蛋白酶体功能下降在帕金森病发病机制中的作用.方法 将蛋白酶体抑制剂立体定向注射入大鼠左侧黑质致密部,对照组注射等体积的生理盐水.观察大鼠自主行为和阿朴吗啡诱导的旋转行为的改变.开野实验观察大鼠自发运动行为改变.免疫组织化学法观察黑质致密部及纹状体内酪氨酸羟化酶的表达.结果 Lactacystin组大鼠给药一周后出现自发性活动减少,动作缓慢,震颤、对外界刺激不自主竖毛,且症状逐步加重;阿朴吗啡可诱导出向健侧的旋转运动.开野实验中,Lactacystin组大鼠自发运动行为出现改变.三周后黑质部位酪氨酸羟化酶免疫阳性细胞及纹状体内酪氨酸羟化酶免疫阳性神经纤维减少.结论 蛋白酶体抑制剂Lactacystin单侧黑质致密部微量注射可以诱导大鼠黑质变性并出现运动行为改变,蛋白酶体抑制剂在帕金森病动物模型制备上有潜在价值,蛋白酶体功能下降可能在帕金森病发病机制中起重要作用.     

Lactacystin诱导的帕金森病大鼠模型病理退行性改变的研究

目的 观察Lactacystin诱导的帕金森病(PD)模型大鼠的病理退行性改变,探索其发病机制.方法 取成年健康SD大鼠32只,采用随机数字表法分为实验组(16只)和对照组(16只),实验组左侧黑质致密部(SNc)注射蛋白酶体抑制剂Lactacystin,对照组以等体积生理盐水代替.分别存注射后1、3、5、7、9、11、14和21 d,观察两组大鼠的行为学变化;应用HE染色观察小胶质细胞并计数,免疫组化检测黑质及纹状体组织罗暗算酪氨酸羟化酶(TH)神经元变化和RT-PCR检测黑质部TH和α-synuclein mRNA的表达.结果 与对照组相比,实验组大鼠逐渐出现行为学改变:HE染色显示小胶质细胞在注射Lactacystin后第1天为(3501.92±57.32)个,第7天为(4895.50±52.67)个,第21天为(5340.18±52.87)个,与对照组(3271.23±63.76)个相比增高,差异具有统计学意义(P＜0.05):免疫组化检测显示黑质中多巴胺(DA)神经元在注射Lactacystin后逐渐变性死亡,第7天神经元数为(568.57±36.39)个,第21天为(119.67±21.06)个,与对照组(679.76±30.24)个相比,差异具有统计学意义(P＜0.05);免疫组化检测纹状体TH免疫阳性纤维显示其在注射Lactacystin后逐渐稀疏,强度逐渐变弱,实验组TH免疫阳性纤维平均光密度第7天为(0.1953±0.0076),与对照组(0.2412±0.0067)相比,差异无统计学意义(P＞0.05),第21天为(0.0781±0.0013)个,与对照组(0.2412±0.0067)个相比降低,差异具有统计学意义(P＜0.05).同时,实验组mRNA检测显示,THmRNA表达越来越少,而α-synuclein mRNA在残存TH神经元中逐渐增多.结论 Lactacystin诱导的PD模型大鼠的病理呈退行性改变.符合PD发病的隐匿性、缓慢进展性特征.     

阻断泛素-蛋白酶体通路诱导PC12细胞死亡和泛素阳性包涵体生成

目的探讨泛素蛋白酶体通路的功能障碍对于多巴胺能细胞的活力以及胞质内包涵体生成的影响。方法应用蛋白酶体抑制剂lactacystin(5μmol/L、10μmol/L、15μmol/L和20μmol/L)处理PC12细胞24h,MTT方法检测细胞活力,WesternBlot方法测定细胞内泛素化蛋白质水平,免疫荧光细胞化学染色观察泛素免疫阳性包涵体的生成。结果经5μmol/L、10μmol/L、15μmol/L和20μmol/Llactacystin处理24h后,PC12细胞的活力显著降低(细胞存活率分别为81.5%±3.6%、75.4%±2.4%、70.2%±2.7%和60.4%±3.9%),呈现剂量依赖性。WesternBlot证实对照组细胞内未检测到相对高分子质量的泛素化蛋白质;随着lactacystin作用浓度的增加,细胞内相对高分子质量泛素化蛋白质的含量逐渐增高。免疫荧光染色显示对照组中仅有极少数细胞内含有泛素阳性包涵体;20μmol/Llactacystin处理组中含有泛素阳性包涵体的细胞数目显著增多(P<0.01)。结论泛素蛋白酶体通路的功能缺失能诱导多巴胺能细胞死亡,造成细胞内泛素化蛋白质积聚,促进胞质内泛素阳性包涵体的生成,可能在帕金森病黑质多巴胺能神经元变性死亡和Lewy小体形成中发挥重要作用。     

体外诱导骨髓基质干细胞治疗帕金森病大鼠模型的研究

目的探讨骨髓基质干细胞(bonemarrowstromalcells,BMSCs)诱导分化为巢蛋白(nestin)阳性的神经干细胞后移植治疗帕金森病动物模型的疗效及作用机制。方法将建模成功的帕金森病SD大鼠模型分为3组,将溴尿嘧啶(brdu)标记的诱导分化的BMSCs(每只鼠共移植6×105个细胞)、未诱导的BMSCs(每只鼠共移植6×105个细胞)和生理盐水(12μl)分别注入模型鼠右侧纹状体,在治疗后1～5个月分别观察阿朴吗啡诱导的旋转行为的变化,并进行脑冰冻切片荧光免疫组织化学鉴定(免疫荧光检测双标细胞)以及脑切片酪氨酸羟化酶(tyrosinhydroxylase,TH)免疫组织化学检测TH阳性细胞并计数。结果细胞移植治疗后1个月,BMSCs诱导组和未诱导组旋转次数开始明显少于生理盐水组[前两组转数分别从移植前的(15.2±2.5)r/min、(14.4±3.8)r/min降到治疗后第5个月的(6.7±1.8)r/min和(8.5±3.2)r/min];而生理盐水组转数移植前后变化不明显,与前两组相比,差异有统计学意义(P<0.05);在移植治疗后,BMSCs诱导组移植区可见一定数量双标的神经和神经胶质细胞,而未发现明显的TH双标的细胞。通过对移植后TH阳性细胞的定量分析可发现:各组间黑质损毁侧TH阳性细胞生存率差异无统计学意义(P>0.05)。结论BMSCs诱导组治疗帕金森病模型的疗效总体好于未诱导的BMSCs组,前两组的疗效均好于生理盐水组。     

鱼藤酮帕金森病大鼠模型蛋白酶体活性的研究

目的研究鱼藤酮对大鼠大脑蛋白酶体活性的影响。方法采用健康Wistar雄性大鼠,建立鱼藤酮帕金森病模型,对实验组和正常对照组大鼠的黑质区和海马区蛋白酶体的活性进行测量,蛋白酶体活性测定通过蛋启酶体与其3种荧光底物反应,采用荧光分光光度计测量反应物荧光强度而定。结果实验组黑质区蛋白酶体 3种活性与对照组相比,蛋白酶体的postacidic活性明显下降(P<0．05),chymotryptic,tryptic活性有极明显下降 (P<0．01);海马区的postacidic活性无明显下降,chymotryptic,tryptic活性有极明显的下降(P<0．01);鱼藤酮处理前后黑质区和海马区3种活性下降程度比较,postacidic活性下降值差异无显著性意义(P>0．05),chymotryptic, tryptic活性下降值差异均有极显著性意义(P<0．01)。结论鱼藤酮对大鼠的黑质区及海马区的蛋白酶体的活性均有损害作用,但对黑质区的损害作用更明显。     

蛋白酶体抑制剂诱导的帕金森病模型中galectin-1含量变化的研究

目的检测蛋白酶体抑制剂PSI诱导的帕金森病(PD)细胞模型中,β半乳糖苷结合血凝素(galectin-1)的含量变化及其在PC12细胞内的分布。方法PSI作用于PC12细胞后,荧光染料染色检测细胞凋亡率;HE染色观察细胞形态学变化。对PSI诱导的PD细胞模型进行蛋白质组学研究,应用免疫细胞化学的方法研究galectin-1在PC12细胞内分布,应用免疫印迹的方法研究此模型中galectin-1的含量变化。结果10μMPSI作用于PC12细胞24小时,细胞凋亡比例增加(P<0.05),胞浆内出现了嗜酸性包涵体。蛋白质组学和免疫印迹研究均提示galectin-1的含量在细胞模型组明显减少(P<0.05)。galectin-1主要分布于PC12细胞的胞膜、胞浆及细胞核内。结论蛋白酶体抑制剂PSI作用于PC12细胞,模拟了人类PD,在此模型中galectin-1减少,提示此蛋白可能在PD的发病机制中起作用。     

Establishing motor disorder mouse models of Parkinson disease——Comparison of 6-hydroxydompamine and l-methyl-4-phenyl-l,2,3,6-tetrahdropyridine

BACKGROUND: At present, pathogenesis and mechanism of Parkinson disease (PD) are still unclear. Animal models of PD are essential tools in studies on etiology and therapy and should mimic the chronic pathological process, histological characteristics and motor behavior dysfunction. In recent years, transgenic mice have been widely utilized to study the mechanism of PD, and it has become imperative that a PD mouse model of motor behavioral dysfunction be established. OBJECTIVE: To compare the behavioral and histochemical characters of two neurotoxic mice model induced with 6-hydroxydopamine (6-OHDA) or 1-methyl-4-phenyl-1, 2, 3, 6 -tetrahydropyridine (MPTP), and a better method to mimic Parkinson disease will be found out. DESIGN: Parallel experiment. SETTING: Laboratory of Molecular Genetics, Department of Medical Genetics, Shanghai Jiao Tong University. MATERIALS: Sixty 129Sv/C57BL6J male wild mice, SPF grade, 8–12 weeks old, weighing 20–25 g, were provided by Experimental Animal Center, Shanghai Jiao Tong University. All the surgery operation was performed according to the rules of Shanghai Jiaotong University Animal Committee. METHODS: The experiment was carried out in the Laboratory of Molecular Genetics (National Key Laboratory), Department of Medical Genetics, Shanghai Jiao Ttong University from March to August 2006. ① Thirty-two male mice were randomly divided into control group and drug treatment group with 16 mice in each group. Surgery was carried out and 6-OHDA was administrated to substantia nigra pars compacta (SNpc) and nigra-striatum pathway according to the different parameters with intoxication apparatus. Saline was injected to the other 16 mice according to the same paradigm. 1 mg/kg apomorphine was injected intraperitoneally 2 weeks later after surgery to induce the imbalanced rotation behavior for 40 minutes. ② Twenty-eight mice were randomly divided into 4 groups with 7 in each group, including low-dose, moderate-dose, high-dose groups and negative control group. Then, mice in the drug treatment group were injected intraperitoneally with 5, 10 and 15 mg/kg MPTP for 9 successive days. In addition, mice in the control group were injected with the same volume of saline for 9 days. Pole test and stride length test were utilized to detect coordinative behavioral dysfunction. Mice were sacrificed 20 days after MPTP treatment, and histochemical staining of tyrosine hydroxynase (TH) was used to observe the loss of dopaminergic neuron in SNpc. MAIN OUTCOME MEASURES: ① Success ratio of each model establishment method; ② inducible asymmetric cycle behavior test 2 weeks after 6-OHDA injection; ③ behavioral dysfunction in pole test and stride length, morphological changes in brain tissue. RESULTS: Totally sixty mice were used in this experiment and 3 mice were excluded because of the hypersensitivity or the clumsy reaction in motor behavioral detection before MPTP treatment, therefore, data was analyzed with the rest 57 mice. ① Lethal ratio: Three out of 16 mice died in striatum injection group and 5 out of 16 mice died in nigro-striatal pathway group. No mouse died in MPTP treatment groups. ② Locomotor behavior: No dysfunction of locomotor was found in 6-OHDA treatment groups. However, several motor behavioral dysfunction were start to present at the 4th day of MPTP injection. ③ Asymmetric cycle behavior: No asymmetric cycle was induced successfully two weeks after 6-OHDA surgery. Mice show hypersensitive behavior 10 minutes after apomorphine injection, which lasted for about 20 minutes. ④ Pole test: From the 4th day of MPTP treatment, mice started to display coordinate dysfunction, such as climbing down along the pole in spiral, moving slowly with hesitation. Some mice could not grab the pole and slide down along the pole at 4th day post injection. Comparing with 0 dose control group, all the three drug treatment groups show significant different dysfunction from the 4th day to the 20th day post injection (P < 0.01). ⑤ stride length test: Mice's stride length decreased, when treated with MPTP, and the mice in the high dose group displayed obviously. ⑥ Dopaminergic neuron stained with TH in nigra pars compacta: The results indicated that administrated MPTP (from low dose to high dose) by intraperitoneal cause chronic lesions on the dopaminergic neuron in the SNpc. CONCLUSION: PD mice models induced with 6-OHDA show high mortality ratio and no asymmetric cycle was found after apomorphine injection. However, injection of MPTP intraperitoneally can simulate the chronic pathway of PD, typical histological changes are found and stable motor behavioral dysfunctions are displayed.     

Dyskinesias and motor symptoms onset in Parkinson disease

PurposeTo assess, using a longitudinal follow-up study design, the relationship between the body site of motor symptoms onset and that of dyskinesias (LID) onset in 70 PD patients in whom LID were absent at the baseline but appeared at one of the follow-up visits; to investigate the demographic and clinical features associated with different sites of LID onset.MethodsMotor symptoms onset was retrospectively determined by asking patients which body part had first been affected by motor impairment. The site of LID onset was determined objectively in one of the follow-up visits.ResultsMotor symptoms started in the limbs in all patients (unilaterally in 91.4% and bilaterally in 8.6% of the patients). LID started unilaterally in the limbs in 25.8%, bilaterally in the limbs in 7.1%, in the cranio-cervical–axial region in 40% and in both the cranio-cervical–axial region and limbs in 27.1% of the patients. No significant association emerged between the site of motor symptoms onset and that of LID onset; a correlation did emerge between the site of motor symptoms onset and that of LID onset in patients with unilateral onset of both motor symptoms and LID. No differences were detected when the subgroups of patients with LID onset in different body regions were compared.ConclusionsThe partial association between the body site of motor symptoms and of LID onset likely reflects pathophysiological mechanisms underlying LID.     

REM sleep behavior disorder in Parkinson disease: Association with abnormal ocular motor findings

BackgroundThe anatomical substrates associated with generalized muscle atonia during REM sleep are located on the pontine tegmentum and medial medulla oblongata. We examined whether patients with REM sleep behavior disorder (RBD) have abnormal ocular movements suggesting brainstem or cerebellar dysfunction in Parkinson's disease (PD).MethodsCross-sectional survey for the existence of RBD and abnormal ocular movements. Ocular movements were examined by video-oculography (VOG).ResultsA total of 202 patients were included in this study. One hundred and sixteen (57.4%) of the 202 patients have clinically probable RBD, and 28 (24.1%) of the 116 with clinically probable RBD patients had abnormal VOG findings suggesting brainstem or cerebellar dysfunction; whereas 86 of the 202 patients did not have clinically probable RBD, and only 7 (8.1%) of the 86 patients had abnormal VOG findings suggesting brainstem or cerebellar dysfunction (P = 0.001).ConclusionThis study suggests that the presence of RBD is associated with more severe or extensive brainstem pathology or different distribution of pathology in PD.     

缝隙连接蛋白36表达变化对帕金森病模型大鼠基底节环路功能的影响

目的 观察缝隙连接蛋白36(Cx36)在帕金森病模型大鼠纹状体和运动皮质区的表达变化,并探讨缝隙连接功能异常与帕金森病基底节环路功能紊乱间的关系。方法 采用6羟多巴胺注射法建立帕金森病动物模型,免疫组织化学染色及Westernblotting法检测纹状体及运动皮质区Cx36表达变化,免疫荧光双标染色进一步分析纹状体脑啡肽阳性传出神经元及Parvalbumin阳性中间神经元Cx36表达变化。结果 (1)免疫组织化学染色显示,帕金森病组大鼠右侧纹状体及运动皮质区Cx36表达水平高于正常对照组(均P<0.05)。(2)免疫荧光双标染色显示,纹状体脑啡肽阳性神经元数目和Cx36表达水平高于正常对照组(均P<0.05),而Parvalbumin阳性神经元数目和Cx36表达水平低于正常对照组(均P<0.05)。(3)Westernblotting法检测显示,帕金森病组大鼠右侧纹状体[(119.31±8.92)%]及运动皮质区[(138.20±17.88)%]Cx36表达水平高于正常对照组[(104.05±3.82)和(105.27±2.82)%,均P<0.05]。结论 帕金森病大鼠右侧纹状体及运动皮质区Cx36表达水平升高,纹状体脑啡肽阳性传出神经元Cx36表达上调,Parvalbumin阳性中间神经元Cx36表达下调。提示缝隙连接异常可能参与帕金森病皮质基底节皮质环路功能紊乱的发生机制。     

Trichloroethylene and Parkinson disease

Multiple genetic and environmental etiologies have been implicated in the pathogenesis of idiopathic Parkinson disease. Recent observations have suggested an association between chronic exposure to trichloroethylene (TCE) and development of clinical parkinsonism. Animal models of TCE exposure have shown nigrostriatal degeneration and the development of parkinsonian features. Animal and cell culture models indicate mitochondrial dysfunction as the probable mechanism, most likely mediated by TaClo, a potential TCE metabolite. These observations endorse the hypothesis that a variety of environmental risk factors may cause nigrostriatal degeneration and clinical parkinsonism in genetically predisposed individuals.     

Orthostatic hypotension predicts motor decline in early Parkinson disease

BackgroundOrthostatic hypotension is increasingly reported as a risk factor for development of late-stage disease features in Parkinson disease (PD). Less is known about its significance in individuals with early PD who are often targeted for neuroprotective trials.MethodsUsing data from the CALM-PD trial (n = 275), we explored whether early orthostatic hypotension predicts a decline in the Unified Parkinson's Disease Rating Scale (UPDRS) II (activities of daily living) or UDPRS III (motor) score after 102 weeks. We also explored risk factors for worsening orthostatic hypotension over a nearly 2-year period.ResultsAfter controlling for age, disease duration, gender, study drug, change in mini-mental status exam score, levodopa equivalent dose, and baseline UPDRS II or III score respectively, the degree of orthostatic hypotension at enrollment associated with a worsening in UPDRS motor score (t = 2.40, p = 0.017) at week 102 but not with UPDRS ADL score (t = 0.83, p = 0.409). Worsening in orthostatic hypotension during the study associated with longer disease duration (t = 2.37, p = 0.019) and lower body mass index (BMI) (t = −2.96, p = 0.003).ConclusionsBaseline orthostatic hypotension is a predictor of UPDRS motor decline in individuals with early PD and should be accounted for in clinical trial design. Low BMI may predict orthostatic hypotension in PD.     

Risk factors for Parkinson disease and the path analysis: One-to-one paired design

BACKGROUND: Parkinson disease (PD) results from the reduce of neurotransmitter dopamine that transmits intracellular information in brain caused by some reasons, then leads to the dynamic disequilibrium with another neurotransmitter of acetylcholine which is relatively hyperactive. The main causes for PD are still unclear. OBJECTIVE: To screen out the risk factors of PD by means of univariate analysis and multivariate Logistic regression analysis, and investigate the manner of actions between various factors and PD, so as to provide clues for the etiological study of PD. DESIGN: A paired design, Logistic regression analysis, path analysis. SETTING: Department of Scientific Research, Shandong Institute of Physical Education. PARTICIPANTS: Totally 157 PD patients were selected from the Department of Neurology, Qilu Hospital of Shandong University from November 2001 to October 2002. Inclusive criteria: PD was diagnosed according to the standard set by the Fourth National Seminar on Extrapyramidal Disease, Parkinsonian syndromes caused by stroke, carbon monoxide poisoning, encephalitis, drugs, etc. were excluded. Another 157 patients treated in the same department at the same period were selected as the control group, they were the same in sex as those in the patient group, within 3 years older or younger than those in the patient group, and without PD or other extrapyramidal diseases. METHODS: ① The general conditions were investigated in all the subjects, including general conditions, social behavioral factor, environmental factor, genetic factor, life events, and previous disease; There were 12 main variables, including educational level, family history, mental labour, contact to insecticides, living place before school-age, smoking index, drinking index, tea-drinking index, history of brain trauma, history of cardiovascular disease, history of diabetes mellitus, and history of depression. ② SAS6.12 software and SPSS 10.0 software were used in the conditional Logistic regression analysis and path analysis respectively. MAIN OUTCOME MEASURES: The results of 12-variable univariate and multivariate analyses; Correlation between main variables and PD; Effects of the factors. RESULTS: All the subjects were involved in the analysis of results. ① The results of Logistic regression analysis showed that family history, mental labour, insecticides, drinking index and history of depression all had significant positive correlations with PD (univariate analysis: OR=1.405–5.429, P < 0.05–0.01; multivariate analysis: OR=2.029–6.754, P < 0.05–0.01), whereas smoking had significant negative correlations with PD [univariate analysis: odd ratio (OR)=0.765, P < 0.05; multivariate analysis: OR =0.489, P < 0.01]. ② The path analysis showed that family history, mental labour, insecticides, smoking, drinking and history of depression had direct effects on PD occurrence [(path coefficient=–0.218 to 0.204, P < 0.05–0.01)]; Insecticides could cause PD indirectly on the basis of family history (genetic susceptibility) (path coefficient=0.946, P < 0.01); Insecticides could also cause PD by drinking (path coefficient=0.165, P < 0.01); Drinking could cause PD indirectly on the basis of family history (path coefficient=0.043, P < 0.01). CONCLUSION: The main risk factors of PD are family history, history of depression, drinking, mental labour and insecticides, whereas the protective factor is smoking. PD attack has genetic susceptibility, insecticides and drinking can cause PD on the basis of PD family history. The risk of PD can be decreased by reducing the occasion for contacting the environmental risk factors.     

Risk factors for Parkinson disease and the path analysis： One-to-one paired design

BACKGROUND： Parkinson disease （PD） results from the reduce of neurotransmitter dopamine that transmits intracellular information in brain caused by some reasons, then leads to the dynamic disequilibrium with another neurotransmitter of acetylcholine which is relatively hyperactive. The main causes for PD are still unclear. OBJECTIVE： To screen out the risk factors of PD by means of univariate analysis and multivariate Logistic regression analysis, and investigate the manner of actions between various factors and PD, so as to provide clues for the etiological study of PD. DESIGN： A paired design, Logistic regression analysis, path analysis. SETTING： Department of Scientific Research, Shandong Institute of Physical Education. PARTICIPANTS： Totally 157 PD patients were selected from the Department of Neurology, Qilu Hospital of Shandong University from November 2001 to October 2002. Inclusive criteria： PD was diagnosed according to the standard set by the Fourth National Seminar on Extrapyramidal Disease, Parkinsonian syndromes caused by stroke, carbon monoxide poisoning, encephalitis, drugs, etc. were excluded. Another 157 patients treated in the same department at the same period were selected as the control group, they were the same in sex as those in the patient group, within 3 years older or younger than those in the patient group, and without PD or other extrapyramidal diseases. METHODS： （1） The general conditions were investigated in all the subjects, including general conditions, social behavioral factor, environmental factor, genetic factor, life events, and previous disease; There were 12 main variables, including educational level, family history, mental labour, contact to insecticides, living place before school-age, smoking index, drinking index, tea-drinking index, history of brain trauma, history of cardiovascular disease, history of diabetes mellitus, and history of depression. （2） SAS6.12 software and SPSS 10.0 software were used in the conditional Logistic regression analysis and path analysis respectively. MAIN OUTCOME MEASURES： The results of 12-variable univariate and multivariate analyses; Correlation between main variables and PD; Effects of the factors. RESULTS： All the subjects were involved in the analysis of results. （1） The results of Logistic regression analysis showed that family history, mental labour, insecticides, drinking index and history of depression all had significant positive correlations with PD （univariate analysis： OR=1.405- 5.429, P 〈 0.05- 0.01; multivariate analysis： OR=2.029- 6.754, P 〈 0.05- 0.01）, whereas smoking had significant negative correlations with PD [univariate analysis： odd ratio （OR）=0.765, P 〈 0.05; multivariate analysis： OR =0.489, P 〈 0.01]. （2） The path analysis showed that family history, mental labour, insecticides, smoking, drinking and history of depression had direct effects on PD occurrence [（path coefficient= - 0.218 to 0.204, P 〈 0.05 -0.01）]; Insecticides could cause PD indirectly on the basis of family history （genetic susceptibility） （path coefficient=0.946, P 〈 0.01）; Insecticides could also cause PD by drinking （path coefficient=0.165, P 〈 0.01） Drinking could cause PD indirectly on the basis of family history （path coefficient=0.043, P 〈 0.01 ）. CONCLUSION： The main risk factors of PD are family history, history of depression, drinking, mental labour and insecticides, whereas the protective factor is smoking. PD attack has genetic susceptibility, insecticides and drinking can cause PD on the basis of PD family history. The risk of PD can be decreased by reducing the occasion for contacting the environmental risk factors.