免疫相关性血细胞减少症患者外周血淋巴细胞线粒体DNA D-LOOP区多态性的研究  

Single Nucleotide Polymorphism of Mitochondrial DNA D-LOOP Region in Peripheral Blood Lymphocytes of Immuno-related Pancytopenia Patients

作　　者：周秋帆 徐淑梅[2] 王化泉[1] 邢莉民[1] 付蓉[1] 邵宗鸿[1] 

机构地区：[1]天津医科大学总医院血液科,天津300052 [2]天津医科大学生理学教研室,天津300070

出　　处：《中国实验血液学杂志》2017年第1期186-191,共6页Journal of Experimental Hematology

基　　金：国家自然基金项目(81370607;81570111;81570106);天津市自然科学基金(14JCYBJC27200)

摘　　要：目的:研究免疫相关性血细胞减少症(IRP)患者外周血淋巴细胞线粒体DNA(mtDNA)D-LOOP区多态性的意义及其与免疫指标的相关性。方法:提取43例初治IRP患者外周血淋巴细胞mtDNA,对其D-LOOP区序列进行PCR扩增,通过正反向直接测序法对PCR扩增产物基因序列进行检测,将检测结果与人线粒体DNA文库记录的剑桥序列以及mtDB数据库的Polmorphic Sites子数据库的相关数据库进行对比。结果:43例IRP患者的外周血淋巴细胞mtDNA D-LOOP区中共有110个变异位点,其中62个为多态位点(SNP),48个为突变位点,其中14个位点为数据库中未提及的新突变位点。在110个变异位点中共发现516个碱基改变,最主要的改变是410个碱基置换,常见的碱基置换为T/C(184/410)和A/G(113/410);110个变异位点中高频变异位点73和263,其变异率为43/43,位点311的变异率为32/43,位点310和16 224的变异率为27/43,16 519的变异率为25/43,位点489和16 362为24/43。对IRP患者外周血淋巴细胞mtDNA D-LOOP区多态性与患者骨髓单个核细胞的相关性统计发现,以年龄18岁为界的35例成年组中淋巴细胞mtDNA D-LOOP区多态性与患者骨髓单个核细胞的CD15 IgM、GLYCoA^+细胞IgM、CD34^+细胞IgG、CD34^+细胞IgM存在显著相关性。结论:IRP患者外周血淋巴细胞的mtDNA D-LOOP区存在高频突变,且在成年组(年龄≥18岁)中与骨髓单个核细胞膜结合自身抗体显著相关,这可能是免疫相关性血细胞减少症发病机制中的重要环节。Objective;To explore the single nucleotide polymorphism（SNP） of mitochondrial DNA（mfDNA） DLOOP region in peripheral blood lymphocytes of immuno-related pancytopenia（IRP） patients and its correlation with immune parameters.Methods;The D-LOOP region in mitochondrial DNA of lymphocytes in peripheral blood mononuclear cells from 43 patients with untreated IRP was detected by polymerase chain reaction（PCR）.The PCR products were sequenced by the pros and cons direct sequencing methods.The sequencing results were compared with the revised Cambridge reference sequence（rCRS） and the Polymorphic Sites of Human Mitochondrial Genome Database.Results：Among total of 110 variant positions of D-LOOP region in 43 patients,62 was SNP sites and 48 was mutation sites,of which 14 were the new mutation sites not yet registered in the database,516 base variations were observed at 110 positions,the most common variations were base substitutions,among them T/C and A/G was 184/410 and 113/410 respectively.In the 110 variant positions,the high frequency variation sites were 73 and 263 for 43/43,311 for 32/43,310 and 16 224 for 27/43,16 519 for 25/43,489 and 16 362 for 24/43.By the analysis of mitochondrial DNA D-LOOP polymorphism and related clinical immunology indicators of the patient＇s lymphocytes,it was found that D-loop in adult patients（age≥ 18 years old） significantly correlated with CD 15 IgM,GLYCoA＋ Cells IgM,CD34＋ CellsIgG,CD34 ＋Cells IgM correlation.Conclusion：The high frequency of polymorphism exists in mitochondrial DNA D-loop region of lymphocytes in IRPA patients,and was significantly correlates with the autoantibodies in bone marrow mononuclear cells in adult patients,which may be associated with the IRP occurrence.

关 键 词：免疫相关血细胞减少症 线粒体DNA(mtDNA) D-LOOP区 多态性 

分 类 号：R552[医药卫生—血液循环系统疾病] R593[医药卫生—内科学]