先天性心脏病与MTHFR基因多态性及环境因素关系的研究
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摘要
目的：(1)探讨5,10-亚甲基四氢叶酸还原酶(MTHFR)基因C677T基因位点突变的分布情况以及突变分布与先天性心脏病(CHD)的关系。(2)了解母亲(MTHFR)基因C677T基因位点突变的分布与子女基因位点突变关联。(3)了解环境因素与CHD的关系,找出其危险因素,为今后预防CHD探讨有效的工作方式和预防模式。
方法：(1)调查100例CHD患者与200例对照,按照1：2配比进行病例对照研究(按照性别、年龄、居住地进行匹配,病例通过临床及彩超进行诊断)。通过聚合酶链反应——限制性片段长度多态性(polymerase chain reaction restricted fragment length polymorphism, PCR-RFLP)并酶切研究儿童及其母亲的MTHFR C677T基因型分布及其差异,探讨MTHFR基因C677T突变是否与CHD有关,C-T等位基因是否会增加儿童CHD的危险性,同时调查父母孕期及孕前所接触的环境因素。(2)设计《幼儿出生质量影响因素调查表》,对CHD儿童及对照儿童父母用采访的方式进行调查,以获取他们是否暴露于相关环境危险因素的资料。(3)对28个CHD的可能的危险因素进行单因素条件Logistic回归分析,并对单因素条件Logistic回归分析中初筛出的16个与CHD有关联的危险因素进行多因素条件Logistic回归分析。并同时进行因子分析,按照因子贡献率的大小,找出公因子。
结果：(1)在CHD病例组MTHFR基因677C→T的突变高于对照组；(2)母亲MTHFR基因677TT型与子代CHD的发生有一定关联。(3)多因素条件Logistic回归分析可以得到的危险因素分别为：妊高症、解热镇痛药、农药、孕早期膳食、蔬菜摄入、负性生活事件、饮酒、儿童基因型,其OR值分别为：29.76、10.194、15.156、0.414、4.69、6.302、2.333、3.474。(4)因子分析可以得到以下公因子：膳食营养因子、负性生活事件因子、感冒相关因子、烟酒因子、保胎相关因子、农药与蔬菜因子、妊娠合并症因子、基因相关因子。
结论：(1)母亲MTHFR基因与子代CHD有一定关联。(2)CHD患者MTHFR基因677C→T的突变与CHD的发生有一定的关联。(3)环境因素与CHD的发生有一定的关系,通过改善环境因素,可能达到预防CHD发生的目的。
Objectives:(1) To investigate the correlationship between 5,10-methylenetetra hydrofolate reductase(MTHFR)'s C677T genetic polymorphism and environmental factors. (2) To study the correlationship between parents'mutations of MTHFR C677T gene and their children's. (3) To assess the environment risk factors of CHDand to discuss the methods to prevent CHD.
Methods:(1)A paired case-control study (case=100 control=200) was used. Cases and controls were paired according to age, sex and living area. Cases were correctly diagnosed by clinical and colored ultrasonic.To study MTHFR C677T genotype distributing and difference between children and his mother by polymerase chain reaction restricted fragment length polymorphism (PCR-RFLP) and enzymatic cleavage.To analyze the chance of mother transmit two alleles of gene C677T of MTHFR to their children. To detective if the change of MTHFR gene C677T C to T is higher than control group. (2)Design The investigate questionnaire of new born baby, Investigate the parents of cases and controls by visitation and interview to obtain the material whether they have contacted the relevant risk factors or not before and during pregnant period。(3)Use simple Conditional Logistic regression analysis to analyze 28 possible risk factors for CHD,and multi-conditional Logistic regression analysis was used to analyze the 16 selected risk factors associated with CHD.In the same time, we use factor analysis, according the proportion of the factor contribution to find the common factors.
Results:(1) In case group, The change of C to T of MTHFR gene's C677T gene is higher than control group; (2)Mother's C677T gene has some relation with children; (3) We can find risk factors from multi-conditional Logistic regression analysis, such as:Pregnancy-induced hypertension syndrome, Antipyretics, Pesticide exposure condition, Early pregnancy meal quality,the amount of vegetable intake,Negative life events,the amount of alcohol taking, the genotype of children. The odds ratio are:29.76、10.194、15.156、0.414、4.69、6.302、2.333、3.474。(4) We can find common factors by factor analysis, they are:Dietary factors, Negative life events factor,i Cold-related factor, smoking and alcohol drinking factors, Baotai relevant factor, pesticide and vegatable factors, pregnancy complications factors, genetic factors.
Conclusion:(1)The mother's MTHFR Gene has some relation to their children. (2)The change of MTHFR C677T Gene is correlated to CHD. (3)There are some correlations between Environmental factors (such as Dietary, infection, negative life events, smoking and alcohol drinking, pesticide exposure, and pregnancy complications) and CHD. It is possible to preventing CHD by improving environmental factors.
方法：(1)调查100例CHD患者与200例对照,按照1：2配比进行病例对照研究(按照性别、年龄、居住地进行匹配,病例通过临床及彩超进行诊断)。通过聚合酶链反应——限制性片段长度多态性(polymerase chain reaction restricted fragment length polymorphism, PCR-RFLP)并酶切研究儿童及其母亲的MTHFR C677T基因型分布及其差异,探讨MTHFR基因C677T突变是否与CHD有关,C-T等位基因是否会增加儿童CHD的危险性,同时调查父母孕期及孕前所接触的环境因素。(2)设计《幼儿出生质量影响因素调查表》,对CHD儿童及对照儿童父母用采访的方式进行调查,以获取他们是否暴露于相关环境危险因素的资料。(3)对28个CHD的可能的危险因素进行单因素条件Logistic回归分析,并对单因素条件Logistic回归分析中初筛出的16个与CHD有关联的危险因素进行多因素条件Logistic回归分析。并同时进行因子分析,按照因子贡献率的大小,找出公因子。
结果：(1)在CHD病例组MTHFR基因677C→T的突变高于对照组；(2)母亲MTHFR基因677TT型与子代CHD的发生有一定关联。(3)多因素条件Logistic回归分析可以得到的危险因素分别为：妊高症、解热镇痛药、农药、孕早期膳食、蔬菜摄入、负性生活事件、饮酒、儿童基因型,其OR值分别为：29.76、10.194、15.156、0.414、4.69、6.302、2.333、3.474。(4)因子分析可以得到以下公因子：膳食营养因子、负性生活事件因子、感冒相关因子、烟酒因子、保胎相关因子、农药与蔬菜因子、妊娠合并症因子、基因相关因子。
结论：(1)母亲MTHFR基因与子代CHD有一定关联。(2)CHD患者MTHFR基因677C→T的突变与CHD的发生有一定的关联。(3)环境因素与CHD的发生有一定的关系,通过改善环境因素,可能达到预防CHD发生的目的。
Objectives:(1) To investigate the correlationship between 5,10-methylenetetra hydrofolate reductase(MTHFR)'s C677T genetic polymorphism and environmental factors. (2) To study the correlationship between parents'mutations of MTHFR C677T gene and their children's. (3) To assess the environment risk factors of CHDand to discuss the methods to prevent CHD.
Methods:(1)A paired case-control study (case=100 control=200) was used. Cases and controls were paired according to age, sex and living area. Cases were correctly diagnosed by clinical and colored ultrasonic.To study MTHFR C677T genotype distributing and difference between children and his mother by polymerase chain reaction restricted fragment length polymorphism (PCR-RFLP) and enzymatic cleavage.To analyze the chance of mother transmit two alleles of gene C677T of MTHFR to their children. To detective if the change of MTHFR gene C677T C to T is higher than control group. (2)Design The investigate questionnaire of new born baby, Investigate the parents of cases and controls by visitation and interview to obtain the material whether they have contacted the relevant risk factors or not before and during pregnant period。(3)Use simple Conditional Logistic regression analysis to analyze 28 possible risk factors for CHD,and multi-conditional Logistic regression analysis was used to analyze the 16 selected risk factors associated with CHD.In the same time, we use factor analysis, according the proportion of the factor contribution to find the common factors.
Results:(1) In case group, The change of C to T of MTHFR gene's C677T gene is higher than control group; (2)Mother's C677T gene has some relation with children; (3) We can find risk factors from multi-conditional Logistic regression analysis, such as:Pregnancy-induced hypertension syndrome, Antipyretics, Pesticide exposure condition, Early pregnancy meal quality,the amount of vegetable intake,Negative life events,the amount of alcohol taking, the genotype of children. The odds ratio are:29.76、10.194、15.156、0.414、4.69、6.302、2.333、3.474。(4) We can find common factors by factor analysis, they are:Dietary factors, Negative life events factor,i Cold-related factor, smoking and alcohol drinking factors, Baotai relevant factor, pesticide and vegatable factors, pregnancy complications factors, genetic factors.
Conclusion:(1)The mother's MTHFR Gene has some relation to their children. (2)The change of MTHFR C677T Gene is correlated to CHD. (3)There are some correlations between Environmental factors (such as Dietary, infection, negative life events, smoking and alcohol drinking, pesticide exposure, and pregnancy complications) and CHD. It is possible to preventing CHD by improving environmental factors.
引文
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10赖杰,韩力苏,宋平.流行性感冒与先天性心脏病发病关系的探讨.预防医学情报杂志.2003,9(2)：148.
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7王惠珊,袁雪,奚一生,等.北京市3岁以下儿童先天性心脏病患病率的抽样调查.中国围产医学杂志,2002,5(3)：217-218.
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12赖杰,韩力苏,宋平.流行性感冒与先天性心脏病发病关系的探讨.预防医学情报杂志.2003,9(2)：148.
13仇小强,覃益敏,谢晓宇,等.1286例先天性心脏病患儿致病因素的调查研究[J].同济医科大学学报,1999,128(5)：399-401.
14王晓明,张国成.B19病毒对先天性心脏病先天感染的探证及电镜观察.第四军医大学学报.2001,22(4)：340-343.
15钱莹莹,陈燕玉,孙玉琴,等.儿童先天性心脏病相关因素分析[J].宁波医学,2000,12(12)：533-555.
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36 Molloy AM, Mills JL, McPartlin J, et al. Maternal and fetal plasma homocysteine concentrations at birth:the influence of folate, vitamin B12, and the 5,10-methylenetetra-hydrofolate reductase 677C-->T variant. Am J Obstet Gynecol.2002,Mar;186(3):499-503.
37 Konrad C, Muller GA, Langer C,et al. Plasma homocysteine, MTHFR C677T, CBS 844ins68bp, and MTHFR G1958A polymorphisms in spontaneous cervical artery dissections. J Neurol.2004 Oct;251(10):1242-1248.
38朱慧萍.5,10-亚甲基四氢叶酸还原酶与神经管畸形国外医学卫生学分册1998.25(5)：264-266.
39闫丽盈李勇,MTHFRC677T基因多态性与先天性心脏病的关系北京大学学报(医学版)2003,35(4)：448-449.
40闫丽盈,李书琴,赵汉杰,等.父母MTHFR基因型与子代先天性心脏病的关系中国疾病控制.2003,7(2)：94-97.
41 Frosst P,Blom HJ, Milos R et al. A candidate genetic risk factor for vascular disease:a common mutation in methylenetetrahydrofolate reductase. Nat Genet.1995 May;10(1):111-113.
42 Molloy AM, Mills JL, Kirke Pn et al. w blood folates in NTD pregnancies are only partly explained by thermolabile 5,10-methylenetetrahydrofolate reductase:low folate status alone may be the critical factor. Am J Med Genet.1998 Jun 30;78(2):155-159.
43 Christensen B, Frosst P, Lussier-Cacan S,et al. Correlation of a common mutation in the methylenetetrahydrofolate reductase gene with plasma homocysteine in patients with premature coronary artery disease. Arterioscler Thromb Vasc Biol.1997 Mar; 17(3):569-573.
44 Kluijtmans LA, van den Heuvel LP,Boers GH,et al. Molecular genetic analysis in mild hyperhomocysteinemia:a common mutation in the methylenetetrahydrofolate reductase gene is a genetic risk factor for cardiovascular disease. Am J Hum Genet.1996 Jan;58(1):35-41.
45 Li D,Pickell L,Liu Y, Wu Q,et al.Maternal methylenetetrahydrofolate reductase deficiency and low dietary folate lead to adverse reproductive outcomes and congenital heart defects in mice. Am J Clin Nutr.2005 Jul;82(1):188-195.
46 Schwahn B, Rozen R. Polymorphisms in the methylenetetrahydrofolate reductase gene:clinical consequences. Am J Pharmacogenomics.2001;1(3):189-201.
47 McLean RR, Karasik D, Selhub J,et al. Association of a common polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene with bone phenotypes depends on plasma folate status.J Bone Miner Res.2004 Mar;19(3):410-418.
48 Allagher PM, Meleady R, Shields DC,et al. Homocysteine and risk of premature coronary heart disease. Evidence for a common gene mutation. Circulation.1996 Nov 1;94(9):2154-2158.
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52 Clarke R. Homocysteine-lowering trials for prevention of heart disease and stroke. Semin Vasc Med.2005 May;5(2):215-222.
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37 Konrad C, Muller GA, Langer C,et al. Plasma homocysteine, MTHFR C677T, CBS 844ins68bp, and MTHFR G1958A polymorphisms in spontaneous cervical artery dissections. J Neurol.2004 Oct;251(10):1242-1248.
38朱慧萍.5,10-亚甲基四氢叶酸还原酶与神经管畸形国外医学卫生学分册1998.25(5)：264-266.
39闫丽盈李勇,MTHFRC677T基因多态性与先天性心脏病的关系北京大学学报(医学版)2003,35(4)：448-449.
40闫丽盈,李书琴,赵汉杰,等.父母MTHFR基因型与子代先天性心脏病的关系中国疾病控制.2003,7(2)：94-97.
41 Frosst P,Blom HJ, Milos R et al. A candidate genetic risk factor for vascular disease:a common mutation in methylenetetrahydrofolate reductase. Nat Genet.1995 May;10(1):111-113.
42 Molloy AM, Mills JL, Kirke Pn et al. w blood folates in NTD pregnancies are only partly explained by thermolabile 5,10-methylenetetrahydrofolate reductase:low folate status alone may be the critical factor. Am J Med Genet.1998 Jun 30;78(2):155-159.
43 Christensen B, Frosst P, Lussier-Cacan S,et al. Correlation of a common mutation in the methylenetetrahydrofolate reductase gene with plasma homocysteine in patients with premature coronary artery disease. Arterioscler Thromb Vasc Biol.1997 Mar; 17(3):569-573.
44 Kluijtmans LA, van den Heuvel LP,Boers GH,et al. Molecular genetic analysis in mild hyperhomocysteinemia:a common mutation in the methylenetetrahydrofolate reductase gene is a genetic risk factor for cardiovascular disease. Am J Hum Genet.1996 Jan;58(1):35-41.
45 Li D,Pickell L,Liu Y, Wu Q,et al.Maternal methylenetetrahydrofolate reductase deficiency and low dietary folate lead to adverse reproductive outcomes and congenital heart defects in mice. Am J Clin Nutr.2005 Jul;82(1):188-195.
46 Schwahn B, Rozen R. Polymorphisms in the methylenetetrahydrofolate reductase gene:clinical consequences. Am J Pharmacogenomics.2001;1(3):189-201.
47 McLean RR, Karasik D, Selhub J,et al. Association of a common polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene with bone phenotypes depends on plasma folate status.J Bone Miner Res.2004 Mar;19(3):410-418.
48 Allagher PM, Meleady R, Shields DC,et al. Homocysteine and risk of premature coronary heart disease. Evidence for a common gene mutation. Circulation.1996 Nov 1;94(9):2154-2158.
49 Strandhagen E, Landaas S, Thelle DS.Folic acid supplement decreases the homoc-ysteine increasing effect of filtered coffee. A randomised placebo-controlled study. Eur J Clin Nutr.2003 Nov;57(11):1411-7.
50 Brown KS, Kluijtmans LA, Young IS,et al.The 5,10-methylenetetrahydrofolate reductase C677T polymorphism interacts with smoking to increase homocysteine.Atheros-clerosis.2004 Jun;174(2):315-322.
51 Christensen B,Frosst P,Lussier-Cacan S et al.Correlation of a common mutation in the methylenetetrahydrofolate reductase gene with plasma homocysteine in patients with premature coronary artery disease. Arterioscler Thromb Vasc Biol.1997 Mar;17(3):569-573.
52 Clarke R. Homocysteine-lowering trials for prevention of heart disease and stroke. Semin Vasc Med.2005 May;5(2):215-222.