CAJ | 학술논문

目的：探讨抗氧化基因NRF2启动子上游CpG岛内408~994 bp位置处的甲基化水平与男性不育之间的关系。方法酚氯仿法手提精子DNA，建立重亚硫酸盐修饰精子DNA后克隆测序法（即 BSP-克隆测序法），选择正常男性及不育中少、弱、畸精症男性患者每份精液样本 DNA的抗氧化基因NRF2的三个片段，每个片段选择15～20个正确的克隆结果并比较其甲基化水平。结果不育男性少、弱精组和正常精液组其抗氧化基因NRF2启动子上游408～994 bp处99%甲基化位点均为非甲基化状态，正常男性精子 DNA 第7个甲基化位点缺失率（18.14%）明显比少弱精组（24.9%）缺失率低（P＜0.05）。结论虽然正常生育男性及不育男性少、弱精患者抗氧化基因NRF2启动子上游408～994 bp区均呈现非甲基化状态但其第7个甲基化位点缺失可能与精子成熟障碍有关。
목적：탐토항양화기인NRF2계동자상유CpG도내408~994 bp위치처적갑기화수평여남성불육지간적관계。방법분록방법수제정자DNA，건립중아류산염수식정자DNA후극륭측서법（즉 BSP-극륭측서법），선택정상남성급불육중소、약、기정증남성환자매빈정액양본 DNA적항양화기인NRF2적삼개편단，매개편단선택15～20개정학적극륭결과병비교기갑기화수평。결과불육남성소、약정조화정상정액조기항양화기인NRF2계동자상유408～994 bp처99%갑기화위점균위비갑기화상태，정상남성정자 DNA 제7개갑기화위점결실솔（18.14%）명현비소약정조（24.9%）결실솔저（P＜0.05）。결론수연정상생육남성급불육남성소、약정환자항양화기인NRF2계동자상유408～994 bp구균정현비갑기화상태단기제7개갑기화위점결실가능여정자성숙장애유관。
Objective To compare the methylation levels of NRF2 gene promoter in sperm cells between the normozoospermia males and oligoasthenoteratozoospermia males. Methods The sperm DNA was extracted by proteinase-K-chloroform method. The bisulfite mix kit was used to process sperm DNA and 15-20 positive clones were selected and analyzed by PCR for sequencing analysis. Results Although the final statistical analysis found that more than 99%of the CPG loci were unmethylated, while loss rate of the seventh methylation loci in normal sperm(18.14%) was much lower than oligoasthenoteratozoospermia group(24.9%), P<0.05. Conclusions Both normozoospermia men and oligoasthenoteratozoospermia patients had largely unmethylated loci in the upstream 408 bp to 994 bp of NRF2 antioxidant gene promoter regions. However, the seventh methylation sites of sperm may be associated with sperm maturation.