肺动脉高压儿童BMPR2基因突变的研究

发布时间：2018-02-13 13:25

  本文关键词： 肺动脉高压 先天性心脏病 家族性肺动脉高压 骨形成蛋白Ⅱ型受体 基因 突变　出处：《山东大学》2012年硕士论文　论文类型：学位论文

【摘要】：骨形成蛋白Ⅱ型受体(BMPR2)是家族性肺动脉高压(FPAH)(?)口特发性肺动脉高压(IPAH)的重要致病因素。国外研究报道家族性、特发性肺动脉高压及先天性心脏病肺动脉高压患者(CHD/PAH)存在BMPR2基因突变。其中,家族性肺动脉高压BMPR2基因突变类型有140余种。我国先天性心脏病合并肺动脉高压患儿是否也存在BMPR2基因外显子突变及国内家族性肺动脉高压患儿是否存在BMPR2基因新的突变位点,罕见报道,为此,本研究旨在探讨先天性心脏病合并肺动脉高压患儿和家族性肺动脉高压患儿是否存在BMPR2基因突变。 第一部分先天性心脏病合并肺动脉高压患儿BMPR2基因新突变 目的：探讨先天性心脏病合并肺动脉高压患儿是否存在BMPR2基因突变。 研究对象和方法 1.研究对象：本研究收集自2011年2月至2011年8月在山东省立医院小儿心脏科及心外科住院的66例先天性心脏病合并肺动脉高压的患儿作为病例组,包括室间隔缺损25例、房间隔缺损5例、动脉导管未闭16例、复杂先天性心脏病20例。其中男39例,女27例,年龄在29天～18岁之间。66名种族、性别、年龄相匹配的健康儿童作为对照组。所有病例和对照均行超声心动图检测,且病例组中有19例经心导管检测证实。 2.方法 (1)采集66例无血缘关系的先天性心脏病合并肺动脉高压的患儿和66名健康儿童外周静脉血,提取基因组DNA,PCR方法扩增BMPR2基因的1-13个外显子编码序列后,对扩增产物纯化后进行测序,测序峰图用Sequencher4.7Demo软件进行分析,与Gene Bank人BMPR2的基因序列进行比对。 (2)BMPR2基因多态性分析：通过χ2检验比较BMPR2基因多态位点等位基因及其基因型在病例组与对照组间频率分布有无统计学意义。 结果： (1)在66例先天性心脏病合并肺动脉高压患儿中检测出1例部分性房室隔缺损并二尖瓣前叶裂、肺动脉高压患儿BMPR2基因外显子8发生错义突变,而其余的病例和对照者均未发现相同的位点发生碱基突变。 (2) BMPR2基因外显子8发生错义突变的患儿为女性,3岁4个月,活动后口唇紫绀1年半；心脏听诊为胸骨左缘2、3肋间可闻及3/6级喷射样收缩期杂音,P2亢进、固定分裂；胸片示心影增大呈梨形,主动脉结缩小,肺动脉段突出；超声心动图检查显示左房内径2.65cm,右房(横径)3.61cm,右室内径2.48cm,主肺动脉内径2.55cm,房室间隔缺损、二尖瓣前叶裂并大量反流,肺动脉收缩压力为38mmHg,诊断为部分性房室隔缺损并二尖瓣前叶裂、肺动脉高压。 (3)在66例先天性心脏病合并肺动脉高压患儿中发现9名患儿和10名健康儿童BMPR2基因编码区有一个单核苷酸多态位点,即c.2811GA,经统计学分析,此多态位点在两组之间差异无统计学意义。 结论： 1.1例部分性房室隔缺损并二尖瓣前叶裂、肺动脉高压患儿BMPR2基因外显子8发生错义突变(p.Val348Ile, c.1042GA),提示BMPR2基因突变可能和先天性心脏病与肺动脉高压形成有关。本研究为国内首次对先天性心脏病合并肺动脉高压患儿进行BMPR2基因突变筛查,且发现存在BMPR2基因突变,该突变位点为一新的突变位点。 2.9名先天性心脏病合并肺动脉高压患儿和10名健康儿童BMPR2基因编码区均发现有一单核苷酸多态位点(c.2811GA),此多态位点可能与先天性心脏病、肺动脉高压形成无关。 第二部分家族性肺动脉高压儿童BMPR2基因新突变 目的：探讨中国汉族家族性肺动脉高压患儿是否存在BMPR2基因突变。研究对象和方法 1.研究对象：本研究收集2011年8月在山东省立医院小儿心脏科住院的一个家族性肺动脉高压家系,家系中成员共8人,男5例,女3例,其中患者1人。先证者经实验室检查、心电图、胸片及胸部CT以排除引起继发性肺动脉高压的疾病。先证者家系成员经体检、心电图、胸片及超声心动图检查,均未见异常。87名健康儿童作为对照组。 2.方法 采集家系中8名成员和87名健康儿童外周静脉血,提取基因组DNA,用PCR方法扩增BMPR2基因13个外显子编码序列后,对扩增产物纯化后进行测序,和GeneBank人的BMPR2基因序列进行比对。 结果： 1.BMPR2基因C347R错义突变可能是与家族性肺动脉高压发生有关的一个新突变位点。 2.先证者的外祖父及其母亲的BMPR2基因第8外显子的第347位密码子发生T→C的置换,使半胱氨酸变为精氨酸,即p.Cys347Arg,二者无肺动脉高压的临床表型。 3.家系其他的成员和对照组BMPR2基因1-13号外显子基因测序结果均未发现异常改变。 4.先证者,男,3岁8月,活动后乏力2月；口唇紫绀,眼睑浮肿,颈静脉怒张,心尖搏动弥散,可触及抬举样搏动。心音低钝,可闻及奔马律,P2亢进。肝肋下4cm,剑下4cm,质韧,双下肢轻度凹陷性水肿。胸片示肺动脉段突出,右房大。胸部CT示肺动脉主干及左右肺动脉增粗,横径约2cm,右心室增大,心包内可见少量液体密度。超声心动图示右心房内径3.37cm,右心室内经2.88cm,右室后壁0.38cm,主肺动脉内径2.46cm,三尖瓣环扩张,并探及中度反流信号,估测肺动脉收缩压86mmHg,临床诊断为原发性肺动脉高压(重度),右心功能衰竭,心功能Ⅳ级。 结论：(?)BMPR2基因C347R错义突变可能是与家族性肺动脉高压发生有关的个新突变位点。
[Abstract]:Bone morphogenetic protein receptor (BMPR2) is a familial pulmonary arterial hypertension (FPAH) (?) and idiopathic pulmonary arterial hypertension (IPAH) is an important pathogenic factor. It was reported that familial idiopathic pulmonary arterial hypertension and congenital heart disease and pulmonary arterial hypertension (CHD/PAH) mutation in BMPR2 deposit genes. Among them, familial pulmonary hypertension BMPR2 gene mutation type. In our country there are more than 140 kinds of congenital heart disease with pulmonary hypertension in children with whether there is BMPR2 gene exon mutation and the familial pulmonary hypertension in children is a novel BMPR2 gene mutation sites, are rarely reported, therefore, this study aims at study of children with congenital heart disease with pulmonary hypertension in children with familial pulmonary hypertension and the presence of BMPR2 gene mutations.
Part 1 new mutation of BMPR2 gene in children with congenital heart disease combined with pulmonary hypertension
Objective: To investigate whether there is a BMPR2 gene mutation in children with congenital heart disease and pulmonary hypertension.
Research objects and methods
1. research subjects: This study collected from February 2011 to August 2011 in Shangdong Province-owned Hospital pediatric cardiology and cardiac surgery, 66 cases of congenital heart disease with pulmonary hypertension were selected as the case group, including 25 cases of ventricular septal defect, atrial septal defect in 5 cases, 16 cases of patent ductus arteriosus, 20 cases of complex congenital heart disease. There were 39 male and 27 female patients, aged 29 days to 18 years between.66 race, gender, age matched healthy children as control group. All cases and controls were measured by echocardiography, and patients in 19 cases confirmed by cardiac catheterization.
2. method
(1) venous blood were collected from 66 unrelated congenital heart disease with pulmonary hypertension in children and 66 healthy children peripheral, genomic DNA, PCR amplification of BMPR2 gene exon 1-13 encoding sequence, the PCR products were sequenced after purification, analysis of the Sequencher4.7Demo software in the sequence figure Bank gene sequence and Gene BMPR2 were compared.
(2) BMPR2 gene polymorphism analysis: Chi square test was used to compare the allele frequencies and alleles of BMPR2 polymorphisms between the case group and the control group, and whether there was statistical significance in the frequency distribution of the polymorphisms of the BMPR2 gene.
Result锛，

本文编号：1508265