【作者】 王雅琴；

【导师】 朱益华；

【作者基本信息】 福建医科大学 ， 眼科学， 2011， 硕士

【摘要】 目的研究福建省一个大型原发性开角型青光眼(POAG)家系的小梁网糖皮质激素诱导反应蛋白基因(MYOC)突变以及该基因突变与表现型之间的关系。对象与方法收集到一个包括5代共144人的POAG家系,其中17人被确诊为POAG,1人因大杯盘比被诊断为可疑患者,其余126人无症状。共采集到12份血样,其中3人为POAG患者,1人为可疑患者,其余8人为正常人。3位患者均已行手术治疗,目前眼压控制不理想。方法:(1)抽取12位POAG患者和正常家系成员的外周血3ml。(2)使用Wizard Genomic DNA Purification试剂盒从12位家系成员外周静脉血中提取和纯化基因组DNA。(3)参照文献所报道的引物序列设计3对特异性引物。(4)对12位家系成员基因组DNA使用聚合酶链反应(PCR)技术分段扩增MYOC基因的3个编码外显子区域,然后将PCR产物纯化并进行正向和反向测序。(5)结合患者临床表现对该家系基因突变与临床表现型之间的关系进行分析。结果(1)在福建省一POAG家系中发现了MYOC基因突变4例,其中3人为POAG患者,1位疑似POAG患者,其余8人为正常人。(2)PCR产物测序发现MYOC基因突变c.G1099A,即Gly367Arg突变,该突变为首次在中国人中发现,导致第367位的甘氨酸突变为精氨酸,从而引起相应蛋白质发生结构与功能改变。结论该家系表现为常染色体显性遗传伴不完全外显。MYOC基因的Gly367Arg突变可能参与了这个大家系的POAG的发病过程,在这个家系中该突变的表型特点是高眼压、大杯盘比和对外科手术治疗不敏感。MYOC基因突变可引起相应蛋白质的结构及功能发生改变,导致POAG的发生。更多还原

【Abstract】 Purpose: To search for the MYOC mutation of primary open-angle glaucoma (POAG) in a large family in Fujian Province and to investigate the relationship of the genotype and the phenotype.Patients: We performed comprehensive ophthalmologic examinations for a primary open-angle glaucoma family of five generations which includes 144 members according to the diagnostic criteria of primary open-angle glaucoma. There are 17 members were confirmed to have POAG , 1 with high cup disc ratio was considered as POAG suspect, and the remaining 126 were asymptomatic. 12 available blood samples were obtained and 3 of them were confirmed to have POAG, 1 was considered as POAG suspect, and the remaining 8 were asymptomatic. All the 3 patients had already accepted surgery therapy and the result is not well.Methods:(1)3ml blood samples were collected from 12 family members including POAG patients and unaffected members.(2)Genomic DNA was extracted and purified from the peripheral leukocytes by using Wizard Genomic DNA Purification kit.(3)3 paires of specific primers were designed according to the references.(4)Genomic DNA of the 12 family members was amplified using polymerase chain reaction. Then the production was purified and sequenced forward and inverse. ( 5 )Investigate the relationship of the genotype and the phenotype according to the clinical symptoms of the patients.Results: (1)We identified one MYOC mutation in 4 of 12 family members and 3 of them were confirmed to have POAG, 1 was considered as POAG suspect, and the remaining 8 were asymptomatic.(2)The MYOC gene mutation c.G1099A , Gly367Arg, was detected by sequencing the PCR production. The transition at codon 367 produced the mutation of glycine to arginine and the structural and functional alteration of the corresponding protein.Conclusions: Gly367Arg mutation of MYOC is likely responsible for the etiology of POAG in this large family. The phenotype of this mutation is characterized by high IOP, high vertical cup-to-disc ratios and insensitivity to surgery. MYOC gene mutation could cause structural and functional alteration of the corresponding protein which is exactly one of the risk factors of POAG.更多还原