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目的:通过比较帕金森病与特发性震颤、头痛患者脑脊液蛋白质谱,筛选和鉴定与帕金森病密切相关的疾病特异性蛋白(disease-specific proteins,DSPs),寻找帕金森病的生物学标志物。方法:以固相pH梯度等电聚焦(IPG-IEF)为第一向,垂直平板十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)为第二向进行脑脊液蛋白质分离;用图像分析软件PDQuest8.0分析电泳图谱,寻找有意义的差异蛋白点;运用MALDI-TOF-MS质谱鉴定,并对其中差异显著蛋白采用酶联免疫吸附实验(ELISA)进行鉴定。结果:帕金森病与特发性震颤患者及头痛对照组患者脑脊液蛋白质谱的比较,发现3个有意义的差异蛋白质点,其中载脂蛋白E(apolipoprotein E,ApoE)差异显著。ELISA鉴定结果表明,与特发性震颤[(3.59±1.07)mg/L]、头痛患者[(3.67±0.71)mg/L]相比,PD组患者脑脊液ApoE水平[(2.47±1.27)mg/L]明显下调,差异有统计学意义(P<0.05)。结论:帕金森病患者与特发性震颤患者、头痛对照组患者脑脊液蛋白表达存在差异,这些差异蛋白可能是帕金森病的DSPs,并有可能成为帕金森病诊断的分子标志物。
OBJECTIVE: To screen and identify disease-specific proteins (DSPs) that are closely related to Parkinson’s disease by comparing the protein profiles of cerebrospinal fluid in patients with Parkinson’s disease and essential tremor and headache, and to search for biomarkers of Parkinson’s disease Things. Methods: IPG-IEF was the first direction, and the vertical plate sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) The image analysis software PDQuest8.0 was used to analyze the electropherogram and search for meaningful differential protein spots. The proteins were identified by MALDI-TOF-MS and the differential proteins were identified by enzyme-linked immunosorbent assay (ELISA). Results: Comparison of cerebrospinal fluid protein profiles between Parkinson’s disease and patients with idiopathic tremor and headache control found three significant differences in protein spots, of which apolipoprotein E (ApoE) was significantly different. The results of ELISA showed that the level of ApoE in cerebrospinal fluid of patients with PD was (2.47 ± 1.27) mg / L compared with [(3.67 ± 0.71) mg / L] patients with idiopathic tremor [(3.59 ± 1.07) mg / L] was significantly decreased, the difference was statistically significant (P <0.05). Conclusions: There are differences in cerebrospinal fluid protein expression between patients with Parkinson’s disease and patients with idiopathic tremor and headache control. These differential proteins may be DSPs of Parkinson’s disease and may become molecular markers for the diagnosis of Parkinson’s disease.