线粒体DNA缺失与线粒体病临床表型复杂程度的关系分析  

Analysis of the relationship between mitochondrial DNA deletion and clinical complexity of mitochondrial disease

作　　者：张彦春[1] 马祎楠[2] 卜定方[2] 刘辉[3] 夏昌宇[4] 张英[2] 朱赛楠[5] 郑雪飞[2] 裴珮[2] 王松涛[2] 许玉凤[2] 潘虹[2] 戚豫[2] 

机构地区：[1]首都医科大学附属北京妇产医院北京妇幼保健院围产保健科,100026 [2]北京大学第一医院中心实验室 [3]北京儿童医院呼吸科 [4]北京大学第一医院检验科 [5]北京大学第一医院统计室

出　　处：《中华医学杂志》2015年第42期3449-3453,共5页National Medical Journal of China

摘　　要：目的 探讨线粒体DNA（mtDNA）4 977 bp缺失（△mtDNA4977）突变比例和拷贝数与线粒体病临床表型复杂程度的关系,为进一步明确病因提供依据.方法 选取2003年12月至2013年12月北京大学第一医院160例线粒体病患者,同时选取101名健康体检者为健康对照组.收集两组患者的外周血DNA样品,经限制性片段长度多态性（RFLP）聚合酶链反应检测,未发现常见的热点突变.将所有患者按起病年龄分为低年龄组（＜10岁）和高年龄组（10岁≤年龄＜20岁）,运用实时定量聚合酶链反应检测缺失突变比例及拷贝数,利用内参核基因计算每个细胞中mtDNA的数量;采用独立t检验、方差分析和Spearman相关分析进行统计.结果 线粒体疾病组的△mtDNA4977突变比例在低年龄组为（2.66±0.63）％,在高年龄组为（3.09±0.74）％,均比健康对照组的同年龄层的突变比例高（t=8.57、4.38,P＜0.01）;平均每个细胞△mtDNA4977拷贝数在低年龄组为（2.79 ±0.50）拷贝,在高年龄组为（2.97±0.48）拷贝,均比健康对照组的同年龄层的突变拷贝数高（t=4.50、-3.67,P＜0.01）;△mtDNA4977突变比例与线粒体病的复杂程度呈正相关（低年龄组r =0.519,P＜0.01;高年龄组r=0.772,P＜0.01）;平均每个细胞的△mtDNA4977拷贝数与线粒体病的复杂程度呈正相关（低年龄组r =0.389,P＜0.01;高年龄组r=0.607,P＜0.05）;平均每个细胞的总mtDNA拷贝数与线粒体病的复杂程度呈负相关（低年龄组r=-0.260,P＜0.01;高年龄组r=-0.430,P＜0.05）.结论 △mtDNA4977突变比例和拷贝数、总mtDNA拷贝数与线粒体病的复杂程度有相关性,其中以△mtDNA4977突变比例最为密切.Objective To analyze the relationship between proportion of mitochondrial DNA 4 977 bp deletion （△mtDNA4977） or copy number in blood and the clinical complexity to find the pathogenesis of mitochondrial disease.Methods A total of 160 patients with mitochondrial disease and 101 healthy controls of Peking University First Hospital from December 2003 to December 2013 were collected in this study.Their peripheral blood showed no hot-point mutation which detected by polymerase chain reaction-restriction fragment length polymorphism.All the patients were divided into younger group （age 〈 10y） and elder group （1 0y ≤ age 〈 20y）.The incidence of △mtDNA4977 was detected by real-time quantitative PCR.Internal gene was used to calculate the number of mitochondrial DNA in each cell.Statistical analysis were carried out by the independent t-test,one-way ANOVA and Spearman＆#39; s bivariate correlation analysis.Results △mtDNA4977 proportion in the younger group was （2.66 ± 0.63） % and in the elder group was （3.09 ± 0.74） %,both of them were higher than that of healthy control group with the same age （the younger group：t =8.57,P 〈 0.01;the elder group：t =4.38,P 〈 0.01）;△mtDNA4977 copy number per cell in the younger group was （2.79 ± 0.50） copy and in the elder group was （2.97 ± 0.48） copy,both of them were higher than that of healthy control group with the same age （the younger group：t =4.50,P 〈 0.01;the elder group：t =-3.67,P 〈 0.01）.The △mtDNA4977 proportion was positively correlated with the complexity of the mitochondrial disease （the younger group：r =0.519,P 〈 0.01;the elder group：r =0.772,P 〈 0.01）.The △mtDNA4977 copy number per cell was positively correlated with the complexity of the mitochondrial disease （the younger group：r =0.389,P 〈 0.01;the elder group：r =0.607,P 〈 0.05）.However,the total mtDNA copy number per cell was negatively correlated with the complexity of the mitochondrial disease （the younger group：r =-0.

关 键 词：基因缺失 线粒体疾病 DNA拷贝数异常 突变 

分 类 号：R596[医药卫生—内科学]