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四氢叶酸甲酯还原酶 C677T

标题、摘要和全文为机器翻译。 英文原文

  • Papitha P Anand1
  • Vijaya Anand Arumugam1
  • Chandra R Samuel2
  1. Department of Human Genetics and Molecular Biology, Bharathiar University, India ROR
  2. Department of Genetics, University of Madras, Taramani, Chennai, India ROR
下载 PDF (218.1 KB) 引用方式 doi:10.5455/umj.20170402052343

摘要

背景:甲基四氢叶酸还原酶是一种重要的酶,在叶酸代谢和DNA甲基化方面起着主要作用. 这项研究的目的是分析不育男子的MTHFR C677T多态性. 研究设计:从58名不育男子中采集血液样本,并采集了55个肥沃的控制对象。 C677T突变分析是通过聚氨酯链式反应-限制片段长度多态化(PCR-RFLP)方法进行的. 结果: 本研究报告显示,在不育男子体内发现了23种同性共聚物、28种异性共聚物、7种TT,而在控制中,异性共聚物变化为38种同性共聚物、17种异性共聚物,但没有发现异性共聚物。 结论: 因此,在不育病人和控制中发现的基因类型和亚麻的频率,但变异亚麻的频率只在不育男子中发现. 只有在不育动物体内才观察到异性激素TT,而在寡光体和异性激素科中都发现了异性激素CT。 研究必须用大量的样本来扩展.

关键词

全文

全文为机器翻译；以英文原文为准。 英文原文

导言

叶酸代谢对基因组的完整性和稳定性具有重要意义,因为它在错误的自由DNA合成,DNA甲基化模式和修复中起着主要作用. (1)因此,在叶酸盐途径的酶中,叶酸盐摄入量或多态性方面的缺陷可能导致异常DNA合成和甲基化。 甲基四氢叶酸还原酶(MTHFR)是甲硫酮和叶酸代谢的重要酶,对DNA合成和甲基化至关重要. MTHFR将5,10-甲基四氢叶酸还原为5-甲基四氢叶酸,使甲基组能与同基基苯乙烯形成甲硫酮. 甲基安非他明最终被转化为S-乙酰甲基安非他明,后者作为DNA甲基化的 " 甲基 " 捐赠者 (2)亚甲苯戊酸氢folate 参与DNA的合成 通过将尿素转化为胸腺 (3)在小鼠的成年睾丸中,MTHFR的活性比其他器官高,代表它在精子生成中的作用 (4, 5)最近的研究确定,几种基因的遗传调控在精子产生和生育方面发挥着重要作用。 (6)因此,MTHFR基因的改变可以改变精子生成过程,使携带者倾向于不孕. (7, 8)。 。 。

人类MTHFR基因的长度为20 kb(20,336 bp),位于1p36.3(OMIM 607093),有11 exons. (9)在MTHFR中已确认的多态性有40多种,但最严重和临床上最显著的常见变体是exon 4中的C677T,在exon 7中的A1298C,但最常见的是MTHFR基因的C677T,已确认影响这种酶的活性. (10–15). C677T变体用马铃薯(A222V, rs1801133)取代阿兰素,它是MTHFR的最常见和最重的形态. (16–21)这种替代导致MTHFR特定活性减少和可热性增加 (22)。 。 。

Homozygous T677T 变体有30%的剩余活性,与C677C 变体相比,Heterozygous C677T 变体有70%的剩余活性。 (23)这种替代还导致同位素和低等离子叶酸水平升高。 C677T多态化的频率因环境和种族而异 (24). 有人建议,由于DNA和RNA分子的合成改变,与MTHFR多态性有关的低水平叶酸盐将成为不孕的原因。 (4, 15)。 。 。

精子的浓度可以通过叶酸和硫酸锌补充而增加,这进一步说明这种途径在精子生成中的重要性。 (14)5-aza脱氧核糖核酸催化低甲基化的研究也支持了这一点,该研究抑制了在肌肉模型中精子细胞与精子细胞的区别。 (25)因此,对C677T多态性的分析对于不育男子来说非常重要,以阐明亚麻黄变种与男性不育之间的联系。

方法

基因组DNA是从患者的外围血液中提取的,使用酚-氯形式-异胺方法控制样品 (26, 27)在热混合器的帮助下,隔离DNA在1x(工作)TE缓冲中溶解. DNA的量化和质量检查是通过对DNA进行分光测量(Nanodrop方法)进行的。 通过使用纳米滴频谱仪(德语:Thermo Science,德国)对核酸样本进行了260nm和280nm的分析.

纯DNA为A260/A280 1.8至2. <1.8的值表示蛋白质和苯酚等离子体. >2的值表示RNA缩合法的可能性. 此外,基因组DNA的完整性是通过在0.8%的甘油胶上运行来评估的. PCR在由DNA 2μl,ADW(自主蒸馏水)7.6μl,前导底物(59 CATC CCTATTGGCAGGTACCC39)组成的薄壁管中进行的总反应体积为20μl,反转0.2μl(59 GGGAGAGAACCAGCGAAC TCAG39) (28, 29) 0.2μl,主调10μl. PCR循环在罗克梯度PCR中进行. PCR放大条件包括:在95°C时饱和度达10分钟,然后在95°C时达到35个饱和度周期30秒,在63°C时达到30秒,在72°C时达到40秒聚合,在72°C时达到7分钟聚合的最后阶段. 放大后,将0.5μl的Hinf I作为限制酶加入0.5μl的放大产物,保存4小时. 4小时后立即将瓶子转移到80°C的水上浴场,并保存20分钟. 最终该产品在3%的甘油胶电泳中运行. 凝胶图像是使用紫外线转光仪查看的。

结果

23 在不育男子体内发现了同性共聚物(Homozygous CC),28种异性共聚物(Hetozygous CT),7种共聚物(TT),而在控制样本中则没有发现同性共聚物(Homozygous CC),17种异性共聚物(Hetozygous CT)的异性共聚物(Homozygous TT). 在属于Azoospermia和oligozoospermia类的7个不育标本中发现了同位素TT的异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异构异

图1:显示RFLP的Gel电泳. C1和C2是缩合物;MI1到MI10是雄性不育样;M-50bp分子标记.
Figure 1. Gel电波显示RFLP. C1和C2是缩合物;MI1到MI10是雄性不育样;M-50bp分子标记.

这些结果也得到了以前进行的研究的元分析的支持。 我们的研究的观察尤其从前两次关于印度人口的研究中引出了有趣之处,这些研究报告了朱和同事在元分析研究中报告的自相矛盾的结果。 调查结果显示,MTHFR C677T多态论与男性不孕症之间存在着联系。 此外,MTHFR C677T与每个基因模型中相当高的异骨化症风险有关。 同时,在大多数遗传模型中没有发现明显增加的寡头动物(OAT)风险。 (30)另一项元分析研究涉及26项关于MTHFR C677T多态论与男性不孕症相关联的研究,研究结论是,MTHFR C677T多态论与男性不孕症风险增加密切相关。 (31)此外,一项研究涉及360名无阻碍性不育男子,并报告了MTHFR C677T与Azoospermia的关系。 (32)。 。 。

结论

基因类型和阿列尔频率被认为与正常对象相比更高. 由于MTHFR基因在对精子起源的调控中起着主要作用,所以alle变体会扰动导致男性不孕的过程. 然而,需要大量样本、设计良好和高质量的流行病学研究,以确认MTHFR C677T多态性和男性不孕症之间的联系。

声明

作者贡献（CRediT）
所有作者对全部 14 项 CRediT 角色贡献均等（Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Software, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing）。
资助

Funding information was not reported in the published version of this article.

利益冲突
A conflict of interest statement was not reported in the published version of this article.
伦理批准
Ethics approval details were not reported in the published version of this article.
数据可用性
A data availability statement was not reported in the published version of this article.

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Anand PP, Arumugam VA, Samuel CR. Methylene Tetrahydrofolate Reductase C677T Gene Polymorphism In Male Infertility. Ulutas Med J. 2017;3(1):1-4. doi:10.5455/umj.20170402052343

Anand, P. P., Arumugam, V. A., & Samuel, C. R. (2017). Methylene Tetrahydrofolate Reductase C677T Gene Polymorphism In Male Infertility. The Ulutas Medical Journal, 3(1), 1–4. https://doi.org/10.5455/umj.20170402052343

Anand, P.P., Arumugam, V.A. and Samuel, C.R. (2017) 'Methylene Tetrahydrofolate Reductase C677T Gene Polymorphism In Male Infertility', The Ulutas Medical Journal, 3(1), pp. 1–4. doi: 10.5455/umj.20170402052343.

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