男性不育患者精子鱼精蛋白表达的研究
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摘要
目的采用荧光定量聚合酶链反应(FQ-PCR)检测成都地区男性不育患者精子鱼精蛋白1(PRM1)和鱼精蛋白2(PRM2)的mRNA表达水平,探讨鱼精蛋白的表达对男性不育症的影响。方法以GAPDH为内对照,采用FQ-PCR检测30例体检健康组、83例少精子症组、61例畸形精子症组患者的精子PRM1和PRM2mRNA表达水平。结果少精子症组的PRM1和PRM2mRNA表达水平显著低于体检健康组和畸形精子症组,差异有统计学意义(P<0.05);畸形精子症组与体检健康组的PRM1和PRM2mRNA表达率比较,差异无统计学意义(P>0.05)。联合检测PRM1和PRM2,可提高对少精子症诊断的灵敏度和阴性预测值。结论 FQ-PCR检测精子PRM1和PRM2的mRNA表达水平,具有操作简便、价格便宜、标本来源容易等优点,联合检测PRM1和PRM2,可提高对少精子症诊断的灵敏度和阴性预测值,成都地区少精子症PRM1和PRM2的mRNA表达量降低,提示与精子发生存在密切关系,有助于临床对男性不育患者进行基因检测、诊断和个体化治疗,以及进行较大规模的流行病学调查。
Objective To detect the express of PRM1 and PRM2 in sperm of the male infertility patients in Chengdu area by fluorescence quantitative polymerase chain reaction and to investigate the correlation between mRNA transcription level of protamines(PRM1 and PRM2)and the male infertility patients.Methods Using GAPDH as internal control,the expression levels of PRM1 and PRM2 in sperm of 30 healthy subjects,83 oligospermia patients and 61 malformed spermatozoa patients were detected by FQ-PCR.Results The express of PRM1 and PRM2 in oligospermia group was significantly lower than physical health group and teratozoospermia group,the difference was statistically significant(P<0.05).There was no difference in the express of PRM1 and PRM2 between teratozoospermia group and physical health group(P>0.05).The sensitivity and negative predictive value were improved by combined detection of PRM1 and PRM2 to the diagnosis of oligozoospermia.Conclusion The sensitivity and negative predictive value were improved by combined detection of PRM1 and PRM2 to the diagnosis of oligozoospermia.It also can be effective in diagnosis of oligozoospermia though the advantages of simple operation,low price and easy sample source.There are low expression of PRM1 and PRM2 in oligozoospermia in Chengdu,suggesting a close relationship with spermatogenesis,which is helpful for clinical detection,diagnosis and individualized treatment of male infertility patients,as well as large-scale epidemiology survey.
Objective To detect the express of PRM1 and PRM2 in sperm of the male infertility patients in Chengdu area by fluorescence quantitative polymerase chain reaction and to investigate the correlation between mRNA transcription level of protamines(PRM1 and PRM2)and the male infertility patients.Methods Using GAPDH as internal control,the expression levels of PRM1 and PRM2 in sperm of 30 healthy subjects,83 oligospermia patients and 61 malformed spermatozoa patients were detected by FQ-PCR.Results The express of PRM1 and PRM2 in oligospermia group was significantly lower than physical health group and teratozoospermia group,the difference was statistically significant(P<0.05).There was no difference in the express of PRM1 and PRM2 between teratozoospermia group and physical health group(P>0.05).The sensitivity and negative predictive value were improved by combined detection of PRM1 and PRM2 to the diagnosis of oligozoospermia.Conclusion The sensitivity and negative predictive value were improved by combined detection of PRM1 and PRM2 to the diagnosis of oligozoospermia.It also can be effective in diagnosis of oligozoospermia though the advantages of simple operation,low price and easy sample source.There are low expression of PRM1 and PRM2 in oligozoospermia in Chengdu,suggesting a close relationship with spermatogenesis,which is helpful for clinical detection,diagnosis and individualized treatment of male infertility patients,as well as large-scale epidemiology survey.
引文
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[12]OLIVA R.Protamines and male infertility[J].Hum Reprod Update,2006,12(4):417-435.
[13]刘小霞,焦海燕.鱼精蛋白与特发性男性不育的研究进展[J].宁夏医学杂志,2013,35(9):874-875.
[14]张亚亚,覃琼玉,林冲,等.亚慢性苯吸入暴露抑制小鼠单倍体精子核蛋白基因的表达[J].温州医科大学学报,2016,46(4):249-253.
[15]MILLER D,BRINKWORTH M,LLES D.Paternal DNA packaging inspermatozoa:More than the sum of its parts?DNA,histones,protamines and epigenetics[J].Reproduction,2010,139(2):287-301.
[16]李俊,刘芳,宋亚曼,等.人鱼精蛋白1多态性与汉族男性生育力的相关性分析[J].中国现代医学杂志,2016,26(13):48-51.
[17]洪丹,楼哲丰,郑九嘉,等.少精子症患者精子鱼精蛋白表达及染色质包装质量[J].温州医科大学学报,2018,48(4):271-274.
[2]杨洋,王树玉.男性不育影响因素的研究进展[J].中国优生与遗传杂志,2011,19(10):131-132.
[3]JIANG W J,SUN H,ZHANG J,et al.Polymorphisms in protamine 1and protamine 2predict the risk of male infertility:a meta-analysis[J].Sci Rep,2015,5(1):153-154.
[4]蒋卫军,张静,夏欣一,等.鱼精蛋白基因多态性与男性不育相关性研究进展[J].中华男科学杂志,2015,21(12):1134-1137.
[5] BRUNNER A M,NANNI P,MANSUY I M.Epigenetic marking of sperm by post-translational modification of histones and protamines[J].Epigenetics Chromatin,2014,7(1):2.
[6] BAO J Q,BEDFORD M T.Epigenetic regulation of the histone-to-protamine transition during spermiogenesis[J].Reproduction,2016,151(5):55-70.
[7]黄悦悦,薛林涛,何冰.精子DNA损伤在辅助生殖中的研究进展[J].中国临床新医学,2016,9(3):261-265.
[8] HA N L,MOZDARANI H,PELLESTOR F.Impact of DNA damage on the frequency of sperm chromosomal aneuploidy in normal and subfertile men[J].Iran Biomed J,2011,15(4):122-129.
[9]邓君,刘蔚,洪华,等.MUC1基因表达与其他肿瘤标志物在乳腺癌诊断中的应用[J].实用医院临床杂志,2016,13(6):23-25.
[10]王健,任华英,丁显平,等.多重荧光PCR在Y染色体微缺失检测中的应用[J].中国优生与遗传杂志,2015,23(8):5-7.
[11]倪梦霞.郅慧杰,刘帅妹等.TP53基因rs1042522单核苷酸多态性与男性不育相关性研究[J].中华男科学杂志,2017,23(2):142-146.
[12]OLIVA R.Protamines and male infertility[J].Hum Reprod Update,2006,12(4):417-435.
[13]刘小霞,焦海燕.鱼精蛋白与特发性男性不育的研究进展[J].宁夏医学杂志,2013,35(9):874-875.
[14]张亚亚,覃琼玉,林冲,等.亚慢性苯吸入暴露抑制小鼠单倍体精子核蛋白基因的表达[J].温州医科大学学报,2016,46(4):249-253.
[15]MILLER D,BRINKWORTH M,LLES D.Paternal DNA packaging inspermatozoa:More than the sum of its parts?DNA,histones,protamines and epigenetics[J].Reproduction,2010,139(2):287-301.
[16]李俊,刘芳,宋亚曼,等.人鱼精蛋白1多态性与汉族男性生育力的相关性分析[J].中国现代医学杂志,2016,26(13):48-51.
[17]洪丹,楼哲丰,郑九嘉,等.少精子症患者精子鱼精蛋白表达及染色质包装质量[J].温州医科大学学报,2018,48(4):271-274.