散发性多系统萎缩与COQ2基因变异相关性报道

发布时间：2018-05-05 17:41

本文选题：多系统萎缩 + COQ2基因　；参考：《大连医科大学》2016年硕士论文

【摘要】：研究背景与目的:多系统萎缩是一种进展性的神经系统变性病,致残、致死率较高,目前发病原因尚不明确,且无可延缓疾病进展的有效治疗方法。近年来越来越多的研究表明多系统萎缩的发病存在遗传相关性,日本的一项大样本研究证明COQ2基因的变异V393A能够增加多系统萎缩患病风险。本实验旨在分析散发性多系统萎缩患者的发病与COQ2基因变异的关系,探讨亚洲人群多系统萎缩病变的病因及治疗方法,提高对多系统萎缩病变的认识。方法:实验分为病例组和对照组,病例组为133例散发性多系统萎缩患者,对照组为200例非神经系统变性病患者(包括51例周围神经病,25例中枢神经系统感染性疾病,23例累及中枢的结缔组织病,19例脑梗死,18例肌肉疾病,12例颈椎、腰椎椎关节强硬,8例神经衰弱症,7例癫痫,5例头痛以及32例其他非神经系统变性病)。实验样本为从上述患者的外周血中提取的DNA样本。我们对病例组COQ2基因外显子1、2、6、7进行聚合酶链式反应扩增,提纯相应的扩增产物,然后对相应外显子进行测序并通过与正常人类COQ2基因序列的对比发现基因多态性核苷酸。运用限制性内切酶片段长度多态性分析以及等位基因特异性聚合酶链式反应对病例组中已发现的变异进行二次检测以确保测序结果的准确性,同时以此两种方法检测对照组中的基因变异。统计学分析方法运用四格表检验比较两组间基因突变类型、例数与多系统萎缩的发病的相关性。结果:在病例组中发现基因变异G21S(c.61G-A)1例,L25V(c.73T-G)7例,P157S(c.469C-T)1例,V393A(c.1178T-C)7例,X422K(c.1264T-A)1例。对照组中发现基因变异G21S(c.61G-A)1例,L25V(c.73T-G)4例,P157S(c.469C-T)0例,V393A(c.1178T-C)11例,X422K(c.1264T-A)0例。病例组中所有变异经限制性内切酶片段长度多态性分析以及等位基因特异性聚合酶链式反应证实是确定存在的。Tsuji S等人曾报道变异V393A为多发于亚洲人种中的基因变异,并且是散发多系统萎缩C型的发病危险因素,可引起线粒内辅酶Q10的合成减少。Sharma M,Jeon BS和Chen YP等人曾报道变异V393A在欧洲、韩国和中国人群的病例对照研究中与多系统萎缩的发病不相关。在我们的实验中,变异V393A在实验组和对照组中无明显差异,不能证明与多系统萎缩的发病相关。Tsuji S等人曾报道在日本散发多系统萎缩患者中发现变异P157S 1例,在我们的病例组中也发现1例,对照组中0例,统计学无明显差异。G21S、L25V、X422K是从未被报道过的变异。G21S和X422K在病例组和对照组中无统计学差异,不能证明与多系统萎缩的发病相关。L25V在病例组和对照组中无统计学差异,而在病例组OPCA患者与对照组中有统计学差异(p=0.04)。结论:定位于COQ2基因上的变异L25V是一个在日本人群中新发现的橄榄脑桥小脑萎缩的遗传易感因素,而之前报道过的变异V393A不能增加多系统萎缩的患病风险。
[Abstract]:Background & AIM: Multi-system atrophy is a progressive neurodegenerative venereal disease with high mortality. At present, the cause of the disease is not clear, and there is no effective treatment to delay the progress of the disease. In recent years, more and more studies have shown that there is a genetic correlation in the pathogenesis of multi-system atrophy. A large sample study in Japan has shown that the mutation of COQ2 gene V393A can increase the risk of multi-system atrophy. The purpose of this study was to analyze the relationship between the pathogenesis of sporadic multisystem atrophy and the variation of COQ2 gene, to explore the etiology and treatment of multisystem atrophy in Asian population, and to improve the understanding of multisystem atrophy. Methods: the experiment was divided into two groups: the case group was 133 patients with sporadic multi-system atrophy. The control group consisted of 200 patients with non-neurotic diseases (including 51 peripheral neuropathy 25 central nervous system infectious diseases 23 central nervous system connective tissue diseases 19 cerebral infarction 18 muscle diseases 12 cervical vertebrae). There were 8 cases of neurasthenia in lumbar vertebrae and 7 cases of epilepsy, 5 cases of headache and 32 cases of other non-neurotic diseases. The experimental samples were DNA samples extracted from the peripheral blood of the patients mentioned above. We amplified the exon 1 of COQ2 gene by polymerase chain reaction and purified the corresponding product. Then we sequenced the corresponding exon and found the polymorphic nucleotides by comparing with the normal human COQ2 gene sequence. Restriction endonuclease fragment length polymorphism (RFLP) and allele-specific polymerase chain reaction (allele-specific polymerase chain reaction) were used to re-detect the mutations found in the case group in order to ensure the accuracy of the sequencing results. At the same time, two methods were used to detect the gene variation in the control group. Statistical analysis method was used to compare the relationship between the type of gene mutation, the number of cases and the incidence of multiple system atrophy between the two groups. Results: in the case group, there were 7 cases of G21S(c.61G-A)1 gene mutation, 7 cases of P157S, c. 469C-TU, 1 case of V393An, c. 1178T-C, 7 cases of X422KN c. 1264T-An, 1 case of P157S, c. 469C-TU, 7 cases of X422KU c. 1264T-An. In the control group, there were 4 cases of G21S(c.61G-A)1 with gene mutation (P157SU, c.469C-TU) and 0 cases of V393Anc. 1178T-C (n = 11) with X422KU c. 1264T-An (n = 0). The results were as follows: (1) in the control group, there were 4 cases of P157SX c. 469C-TU. All mutations in the case group were confirmed by restriction endonuclease fragment length polymorphism analysis and allele-specific polymerase chain reaction. Tsuji S et al reported that the mutation V393A was a gene mutation in Asian species. It is also a risk factor for sporadic multisystem atrophy type C, which may cause a decrease in the synthesis of coenzyme Q10. Sharma M Jeon BS and Chen YP have reported that variant V393A is present in Europe. Case-control studies in Korean and Chinese populations were not associated with multiple-system atrophy. In our experiment, there was no significant difference between the experimental group and the control group. Tsuji S et al reported that one case of variant P157S was found in patients with sporadic multisystem atrophy in Japan. In our case group, one case was also found, and there was no significant difference between control group and control group. There was no significant difference between the two groups. There was no statistical difference between the case group and the control group, and there was no statistical difference between the case group and the control group. There was no significant difference in the incidence of multisystem atrophy. L25V was not significantly different between the case group and the control group, but there was a significant difference between the case group and the control group in the OPCA patients and the control group. Conclusion: the mutation L25V located on the COQ2 gene is a new genetic susceptibility factor for olivary pontine cerebellar atrophy in Japanese population, while the previously reported mutation V393A does not increase the risk of multiple system atrophy.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R741

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