精子发生障碍患者Y染色体AZF区拷贝数变异的分析

Analysis of copy number variation in AZF region of Y chromosome in patients with spermatogenic failure

目的探讨深圳地区精子发生障碍患者的Y染色体无精子因子(AZF)区域内的拷贝数变异(CNV)特征。方法选取2016年1月至2022年10月就诊于深圳市人民医院的123例精子发生障碍患者(73例无精子症患者,50例少精子症患者)及100例精液正常的男性为研究对象。应用多重连接探针扩增(MLPA)技术检测其AZF区,利用χ2检验或者Fisher确切概率法统计分析AZF区的CNV与男性精子发生障碍的关系。结果223例样本中共检出19种CNV,合计53例,其中无精子症中检出20例(27.40%,20/73),少精子症中检出19例(38%,19/50),正常对照组中检出14例(14%,14/100)。无精子症组、少精子症组及正常对照组中,AZFa区(含AZFab区与AZFabc区)相关的CNV检出率分别为5.48%(4/73)、2.00%(1/50)与0(0/100);AZFb区(含AZFbc区)的检出率分别为6.85%(5/73)、0(0/50)与0(0/100);AZFc区gr/gr缺失检出率分别为2.74%(2/73)、6.00%(3/50)、9.00%(9/100);AZFc区b2/b4缺失检出率分别为2.74%(2/73)、10.00%(5/50)和0(0/100);AZFc区复杂重排检出率分别为6.85%(5/73)、18.00%(9/50)和3.00%(3/100);统计学分析显示,gr/gr缺失在三组间检出率差异无统计学意义(Fisher′s Exact Test值=2.712,P=0.249);b2/b4缺失在三组间检出率差异有统计学意义(Fisher′s Exact Test值=9.489,P=0.002),AZFc区复杂重排在三组间检出率差异有统计学意义(Fisher′s Exact Test值=9.493,P=0.006)。本研究还检出1例罕见的AZFa区ARSLP1基因缺失(涉及SY86缺失)的少精子症患者。结论AZFa区与AZFb区的CNV对男性的精子发生有严重影响,但AZFa区部分缺失(ARSLP1基因缺失)对男性的精子发生影响较小。AZFc区的b2/b4缺失与复杂重排可能为男性不育的风险因子。gr/gr缺失可能无法作为深圳地区男性不育的风险因子。

Objective To explore the characteristics of copy number variation(CNV)within the Y chromosome azoospermia factor(AZF)region in patients with spermatogenesis disorders in the Shenzhen area.Methods A total of 123 patients with spermatogenesis disorders who had visited Shenzhen People′s Hospital from January 2016 to October 2022(including 73 patients with azoospermia and 50 patients with oligozoospermia)and 100 normal semen males were selected as the study subjects.The AZF region was detected with multiplex ligation-dependent probe amplification(MLPA),and the correlation between the CNV in the AZF region and spermatogenesis disorders was analyzed using the chi-square test or Fisher′s exact test.Results 19 CNV were detected among 53 patients from the 223 samples,including 20 cases(27.40%,20/73)from the azoospermia group,19 cases(38%,19/50)from the oligozoospermia group,and 14 cases(14%,14/100)from the normal control group.In the azoospermia,oligozoospermia,and normal control groups,the detection rates for CNV related to the AZFa region(including AZFab and AZFabc)were 5.48%(4/73),2.00%(1/50),and 0(0/100),respectively.The detection rates for the AZFb region(including the AZFbc region)were 6.85%(5/73),0(0/50),and 0(0/100),respectively.The detection rates for gr/gr deletions in the AZFc region were 2.74%(2/73),6.00%(3/50),and 9.00%(9/100),respectively,and those for b2/b4 deletions in the AZFc region were 2.74%(2/73),10.00%(5/50),and 0(0/100),respectively.The detection rates for complex rearrangements in the AZFc region were 6.85%(5/73),18.00%(9/50),and 3.00%(3/100),respectively.Statistical analysis showed no significant difference in the detection rate of gr/gr deletions between the three groups(Fisher′s Exact Test value=2.712,P=0.249);the differences in the detection rate of b2/b4 deletions between the three groups were statistically significant(Fisher′s Exact Test value=9.489,P=0.002);the differences in the detection rate of complex rearrangements in the AZFc region between the three groups were statistically significant(Fisher′s Exact Test value=9.493,P=0.006).In this study,a rare AZFa region ARSLP1 gene deletion(involving SY86 deletion)was detected in a patient with oligozoospermia.Conclusion CNV in the AZFa and AZFb regions have a severe impact on spermatogenesis,but partial deletion in the AZFa region(ARSLP1 gene deletion)has a minor impact on spermatogenesis.The b2/b4 deletion and complex rearrangement in the AZFc region may be risk factors for male infertility.The gr/gr deletion may not serve as a risk factor for male infertility in the Shenzhen area.