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

发布时间：2018-05-28 17:29

本文选题：视神经脊髓炎 + 水通道蛋白质　；参考：《中国全科医学》2017年03期

【摘要】：目的从基因水平研究水通道蛋白4(AQP4)基因外显子区域单核苷酸多态性(SNP)位点以及其在视神经脊髓炎发病机制中的可能作用。方法选取2010年3月—2012年6月湖北医药学院附属襄阳医院收治的视神经脊髓炎患者72例为视神经脊髓炎组,同时期本院收治的多发性硬化患者80例为多发性硬化组,取全血DNA进行AQP4基因的SNP位点分析,并通过特异性的定点诱变技术对携带AQP4基因表达序列的绿色荧光蛋白表达载体(p EGFP)重组质粒进行定点诱变,根据相应SNP位点突变生成相应突变质粒,脂质体转染法将相应质粒转染细胞建立抗AQP4抗体检测细胞株。分析视神经脊髓炎患者6种突变细胞株与原始细胞株血清抗AQP4抗体滴度水平及视神经脊髓炎组与多发性硬化组不同细胞株血清抗AQP4抗体滴度阳性率的差异。结果视神经脊髓炎组共发现6个AQP4基因SNP位点,位于2号和5号外显子,分别为R108T、I110N、E280R、D281R、P295R、E317M。1、3、4号外显子区域未发现AQP4基因SNP位点,多发性硬化组未发现相应的AQP4基因SNP位点。构建R108T、I110N、R108T/I110N、E280R/D281R、P295R、E317M细胞株,R108T/I110N、E280R/D281R、E317M细胞株与原始细胞株血清抗AQP4抗体滴度水平比较,差异有统计学意义(P0.05)。视神经脊髓炎组与多发性硬化组中原始细胞株及R108T、I110N、R108T/I110N、E280R/D281R、P295R、E317M细胞株血清抗AQP4抗体滴度阳性率比较,差异均有统计学意义(P0.05)。结论视神经脊髓炎患者AQP4基因SNP位点可能会导致表达蛋白一级结构发生变化,可以改变其抗原性,抗原抗体反应的强度不同可能是导致不同突变组间滴度差异的原因。
[Abstract]:Objective to study the single nucleotide polymorphisms (SNPs) in the exon of aquaporin 4AQP4 gene and its possible role in the pathogenesis of optic neuromyelitis. Methods from March 2010 to June 2012, 72 patients with optic neuromyelitis and 80 patients with multiple sclerosis treated in Xiangyang Hospital of Hubei Medical College were selected. The whole blood DNA was used to analyze the SNP locus of AQP4 gene, and the recombinant plasmid carrying the expression sequence of AQP4 gene was mutated by specific site-directed mutagenesis technique. The corresponding plasmids were generated according to the mutation of the corresponding SNP sites, and the corresponding plasmids were transfected into the cells by liposome transfection method to establish the anti AQP4 antibody detection cell lines. The titer of anti AQP4 antibody in serum of 6 kinds of mutant cell lines and primordial cell lines in patients with optic neuromyelitis and the titer of anti AQP4 antibody in different cell lines of optic neuromyelitis group and multiple sclerosis group were analyzed. Results six SNP loci of AQP4 gene were found in optic neuromyelitis group, located at exon 2 and exon 5, respectively, which were R108TTHI110NU E280RD280RP295RP295RP295RP295RP317M.1t3, AQP4 gene SNP locus was not found in exon 4, and AQP4 gene SNP locus was not found in multiple sclerosis group. The titer of serum anti AQP4 antibody of R108T / I110NNtr / I110NCU E280R / D281RP295RP317M cell line was significantly higher than that of the original cell line. The titer of serum anti AQP4 antibody of R108T / I110NNC280R / D281RP317M cell line was significantly higher than that of the original cell line (P0. 05, P < 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05). There were significant differences in the titer of serum AQP4 antibodies between the optic neuromyelitis group and the multiple sclerosis cell lines, R108T / I108Tr / I10NP280R / D281RP295RP317M cell lines. Conclusion the AQP4 gene SNP site may lead to the change of the primary structure of the expressed protein in patients with optic neuromyelitis and may change its antigenicity. The different intensity of antigen-antibody reaction may be the cause of the difference of titer among different mutation groups.
【作者单位】：湖北医药学院附属襄阳医院神经内科;贵州医科大学附属医院神经内科;
【基金】：贵州省科技厅科技计划项目(黔科合LG字[2011]004号)
【分类号】：R744.52

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