中国全科医学2017，Vol.20Issue(3)：331-336,6.DOI:10.3969/j.issn.1007-9572.2017.03.013
视神经脊髓炎患者水通道蛋白4基因单核苷酸多态性位点的研究
Single Nucleotide Polymorphic Site of Aquaporin-4 Gene in Patients with Neuromyelitis Optica
摘要
Abstract
Objective To study the single nucleotide polymorphic ( SNP ) site in exon region of aquaporin ―4 (AQP4)gene and its possible effects on the pathogenesis of neuromyelitis optica(NMO)based on gene level. Methods 72 patients with NMO who were treated in Xiangyang Hospital Affiliated to Hubei University of Medicine from March 2010 to June 2012 were selected as NOM group,and 80 patients with multiple sclerosis admitted to this hospital during the same period were treated as multiple sclerosis group. The SNP site analysis of AQP4 was made to the whole blood DNA, and site ― directed mutagenesis was conducted to the recombinant plasmid of plasmid enhanced green florescence protein ( pEGFP ) that carried AQP4 gene expression sequence by specific site ― directed mutagenesis. The corresponding mutated plasmids were generated according to mutation of relevant SNP sites,and lipofection transfection method was used to build anti―AQP4 antibodies of the corresponding plasmid transfected cells so as to detect the cell line. The difference of serum antibody titers level of anti―AQP4 between six mutated cell lines and original cell lines in patients with NMO and positive rate of antibody titer of serum anti―AQP4 of different cell lines between NMO and multiple sclerosis groups were both analyzed. Results Six SNP sites of AQP4 gene were found in NMO group, which were located in exon 2 and 5, they were R108T, I110N, E280R, D281R, P295R and E317M. SNP sites of AQP4 gene were not found in exon 1,3 and 4,and no corresponding SNP sites of AQP4 gene was found in the multiple sclerosis group. Builded the R108T,I110N,R108T/I110N,E280R/D281R,P295R,E317M cell lines. There was significant difference in the antibody titer level of serum anti―AQP4 between the R108T/I110N,E280R/D281R,E317M and the original cell lines(p〈0. 05). There was significant difference in the positive rate of serum anti―AQP4 antibodies of the original cell lines and cell lines R108T,I110N,R108T/I110N,E280R/D281R,P295R,E317M between NMO group and multiple sclerosis group(p〈0. 05). Conclusion The SNP sites of AQP4 gene in patients with NMO may lead to changes in the primary structure of expressing protein and can change its antigenicity. The different intensity of the antigen―antibody reaction may be the reason of the difference in titers between mutation groups.关键词
视神经脊髓炎/水通道蛋白质4/多态性,单核苷酸Key words
Neuromyelitis optica/Aquaporin 4/Polymorphism,single nucleotide分类
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王青松,戴庆箐,王浩,徐竹,张艺凡,楚兰..视神经脊髓炎患者水通道蛋白4基因单核苷酸多态性位点的研究[J].中国全科医学,2017,20(3):331-336,6.基金项目
贵州省科技厅科技计划项目 （）
