脑脊液CystatinC改变与格林-巴利综合征关系的探讨  

Relation between alteration of Cystatin C in cerebrospinal fluid and Guillain-Barre′syndrome

作　　者：杨银荣[1] 刘师莲[1] 刘芙君[2] 秦延江[3] 柏淑美[1] 

机构地区：[1]山东大学医学院生物化学与分子生物学研究所,济南250012 [2]青岛大学医学院烟台毓璜顶医院中心实验室,山东烟台204000 [3]山东大学齐鲁医院神经内科,济南250012

出　　处：《山东大学学报（医学版）》2008年第1期22-26,共5页Journal of Shandong University:Health Sciences

摘　　要：目的对格林-巴利综合征(GBS)患者和对照组脑脊液进行蛋白质组学分析,寻找改变最明显的蛋白质,初步探讨其与GBS的关系。方法分别取GBS疾病组和对照组的新鲜脑脊液,冰丙酮沉淀法提取总蛋白,进行第一向等电聚焦电泳(IEF)和第二向十二烷基磺酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分离蛋白质组,Image Master 2D图像分析软件对分析胶进行比较分析,识别差异表达明显的蛋白质点。抠取制备胶上相应的蛋白质点,应用基质辅助激光解吸电离飞行时间质谱(MALDI/TOF/MS)得到肽质量指纹图谱(PMF),搜索SWISSPROT数据库鉴定蛋白质点。结果双向电泳图谱上有6个点在两组间存在明显差异,4个蛋白质点在GBS疾病组中表达明显上调,2个降低。其中Cystatin C表达下调,是改变最明显的蛋白质点,为本实验首次发现。随后的酶联免疫吸附实验(ELISA)也证实GBS患者的脑脊液中Cystatin C的浓度明显低于对照组[(2.79±0.65)mg/L,(3.65±0.82)mg/L,P<0.01]。结论Cystatin C浓度在GBS患者脑脊液中的浓度明显降低,可为研究格林-巴利综合征疾病机制提供线索。Objective Proteomic analysis was carried out in the cerebrospinal fluid （CSF） of patients with Guillain-Barre＇syn- drome（GBS） and of controls to find the protein presenting the most obvious changes and to investigate its relation with Guillain- Barre＇syndrome. Methods CSF was extracted from patients with GBS and controls, and proteins were precipitated with ice-cold acetone. Isoelectrie focusing electrophoresis（IEF） was employed as the fn＇st dimension and sodium dodecylsulfate-polyacrylamide gel electrophoresis （SDS-PAGE） was employed as the second dimension. The two-dimeusional analysis gel maps were analyzed by Imagemaster 2D software to find the obvious alteration proteins and attain the proteins in the prepared gels. The peptides mass fingerprint was obtained by MOLDI/TOF/MS, and differential proteins were identified in the SWISSPROT database. Results There were 6 protein spots that had significant differences in expression between the two groups. Expressions of 4 protein spots in the GBS group increased and 2 decreased. Cystatin C which was the most significantly decreased protein was firstly discovered in this study. The subsequent ELISA eonfu＇med that the concentration of Cystatin C was significantly lower in the GBS group than that in the control group [ （2.79± 0. 65） vs （3.65 ± 0. 82 ） mg/L, P 〈0. 01 ]. Conclusion That the level of Cystatin C significantly decreases in the CSF of GBS may offer clues for studies on the mechanisms of GBS.

关 键 词：格林-巴利综合征 脑脊髓液 蛋白质组 酶联免疫吸附测定 半胱氨酸蛋白酶抑制剂C 

分 类 号：R446.14[医药卫生—诊断学]