【作者】 陈岳明； 王莉； 裘春宁； 屠巧凤； 梅瑾； 张闻； 钱霞； 王贤军； 罗丽；

【机构】 杭州市第一人民医院浙江省杭州市共建医学重点学科； 温州医科大学；

【摘要】 目的探讨叶酸代谢主要酶编码基因MTHFR 677C>T,MTHFR 1298A>C,MTR 2756A>G,MTRR 66A>G和RFC-1 80A>G多态性与复发性流产(RPL)的关联。方法随机选取杭州地区125例有≥2次RPL史孕妇为病例组,135例生过≥1个健康婴儿且无流产史及其它明显疾病的育龄妇女为对照组。MTHFR 677C>T,MTHFR 1298A>C,MTR 2756A>G和RFC-1 80A>G基因型及等位基因频率分布采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测,并对部分结果行测序验证;MTRR 66A>G多态性采用PCR后直接测序法检测。比较病例组与对照组间基因型、等位基因频率分布及单倍型情况,分析叶酸代谢主要酶编码基因多态性与RPL的关联。结果 MTHFR 677C>T基因型和等位基因频率在病例组分布分别为CC(32.0%)、CT(56.0%)、TT(12.0%)、C(60%)和T(40%),在对照组分别为CC(44.5%)、CT(48.1%)、TT(7.4%)、C(68.5%)和T(31.5%);等位基因C、T在两组间分布频率差异有统计学意义,病例组T等位基因分布频率显著高于对照组(OR=1.451,95%CI=1.012~2.081,P=0.043),病例组(CT+TT)基因型分布频率显著高于对照组(OR=1.700,95%CI=1.024~2.821,P=0.039),两组CC、CT、TT各基因型分布无统计学意义。MTR2756G为RPL的保护因素(OR=0.509,95%CI=0.272~0.952,P=0.032)。两组间MTHFR 1298A>C,MTRR 66A>G,RFC-1 80A>G基因型和等位基因频率分布差异无统计学意义。单倍型分析结果显示677C/1298A/2756A/66A/80A降低RPL发生风险(OR=0.237,95%CI=0.156~0.361,P<0.001),而677T/1298A/2756A/66A/80G增加RPL发生风险(OR=5.749,95%CI=2.997~11.029,P<0.001)。结论 MTHFR 677T和单倍型677T/1298A/2756A/66A/80G为RPL的危险因素,而MTR 2756G和单倍型677C/1298A/2756A/66A/80A为RPL的保护因素。更多还原

【Abstract】 Objective:To explore genes polymorphisms of major enzymes in folate metabolic pathway,including MTHFR 677C>T,MTHFR 1298A>C,MTR 2756A>G,MTRR 66A>G and RFC-1 80A>G associated with recurrent pregnancy loss(RPL).Methods:We selected 125 cases with a history of ≥ 2 times RPL,and 135 healthy fertile women with a history of ≥ 1 times successful pregnancy outcome as control.Polymerase chain reaction- restriction fragment length polymorphism(PCR-RFLP) was used to detect the distributions of genotypes and alleles in MTHFR,MTR and RFC-1,some of which were selected randomly by sequencing to validate.The polymorphism of MTRR was determined by sequencing.The differences in the frequencies of genotypes and alleles of 5 loci between case and control were measured by Chi-square test.In addition,haplotypes were analyzed by SHEsis,one of online biological software.Results:The frequencies of genotypes and alleles of MTHFR 677C>T were CC(32.0%),CT(56.0%),TT(12.0%),C(60.0%),T(40.0%) in case group and CC(44.5%),CT(48.1%),TT(7.4%),C(68.5%),T(31.5%) in control group,respectively;Comparing allele C with T between the two groups,the prevalence of allele T was obviously higher in case group than in control group(OR=1.451,95%CI =1.012 ~ 2.081,P=0.043);the frequency of genotype(CT + TT) was significantly higher in case group than that in control group(OR=1.700,95%CI =1.024 ~ 2.821,P=0.039),whereas there showed no difference among CC,CT and TT genotypes of MTHFR 677C>T.MTR 2756 G showed a protective effect on RPL(OR=0.509,95%CI=0.272 ~ 0.952,P=0.032).In addition,no differences were found between case and control in the frequencies of alleles and genotypes of MTHFR 1298A>C,MTRR 66A>G and RFC-1 80A>G.Haplotype analysis revealed 677T/1298A/2756A/66A/80 G could increase the risk of RPL(OR=5.749,95%CI=2.997 ~ 11.029,P<0.001).Conclusion:MTHFR 677 T and haplotype 677T/1298A/2756A/66A/80 G was a risk factor for RPL,MTR2756 G and haplotype 677C/1298A/2756A/66A/80 A provided protect effect on susceptibility to RPL in fertile women in Hangzhou,China.更多还原