弱活力精子AKAP82mRNA的表达及意义
摘要
目的:通过测定A激酶锚定蛋白AKAP82mRNA在弱精子症患者及健康成年男性精子中的差异,了解其AKAP82mRNA是否存在异常,探讨其在弱精子症发病机理中可能的作用。
方法:采用计算机辅助精液分析系统检测59例弱精子症患者(实验组)和28例健康成年男性(对照组)精液中精子的运动参数,用ELISA方法检测实验组患者血清抗精子抗体(AsAb)。使用低渗肿胀(HOS)试验对两组精子进行分析,通过RT-PCR方法对两组精子中AKAP82mRNA进行检测,实验组患者经提高精子活力活率、抗体专阴治疗后再用RT-PCR方法对其精子AKAP82mRNA进行检测和分析。
结果:①59例实验组(17例AsAb阳性,42例AsAb阴性)中有15例标本在RT-PCR检测中有发现有AKAP82mRNA表达缺失,28例对照组中发现2例有AKAP82mRNA表达缺失。有统计学意义(P＜0.05)。其中AsAb阳性组中有4例AKAP82mRNA表达缺失;AsAb阴性组中有11例AKAP82mRNA表达阴性,并无显著性差异(P＞0.05)。②低渗肿胀(HOS)试验中实验组精子尾部总肿胀率,g型精子率均低于对照组(P＜0.05);实验组中15例RT-PCR检测AKAP82mRNA缺失标本精子尾部总肿胀率,g型精子率低于实验组中AKAP82mRNA未缺失标本(P＜0.05)。③15例AKAP82mRNA缺失的患者经提高精子活力活率、抗体专阴治疗后RT-PCR方法检测发现AKAP82mRNA仍然缺失。
结论:弱精子症患者AKAP82mRNA表达缺失高于健康成年男性;AsAb阳性患者与阴性组AKAP82mRNA表达无显著差异;弱精子症患者低渗肿胀(HOS)试验精子尾部总肿胀率g型精子率低于健康成年男性;在弱精子症患者中AKAP82mRNA表达缺失患者HOS精子尾部总肿胀率g型精子率低于AKAP82mRNA表达阳性患者。
部分弱精子症患者精子的AKAP82异常可能是精子活力低下的原因之一。AKAP82mRNA缺失是造成AKAP82异常表达的原因。
Objective:AKAP82(A-kinase anchor proteins82) is one of A-kinase anchor proteins in sperm.In order to know the AKAP82mRNA levels of the patients with asthenospermia,we examinated the expressions of AKAP82mRNA in the patients and the healthy fertile men.
Methods:Sperm samples were obtained from 59 patients with asthenospermia (experimental group),and the normal sperms were obtained from 28 healthy fertile men(control group).And we estimated motility parameters with a computer-assisted semen analysis(CASA) technique.AsAb(antisperm antibody) of blood serum in the experimental group's was measured by ELISA(enzyme-linked immunosorbentassay). We measured the two groups AKAP82mRNA by reverse transcriptase PCR.At last, after the experimental group was treated,we measured the levels of AKAP82mRNA by reverse transcriptase PCR.
Results:Among the 59 patients,15 patients had been found with AKAP82mRNA microdeletion,and among the 28 healthy fertile men,2 had been found with AKAP82mRNA microdeletion,the percentage of g type sperm and the total tail swelling rate were significantly different(P＜0.05) between control group and experimental group.
Conclusion:Abnormality of AKAP82 expression in sperm may be one of the etiological factors in asthenospermia.Abnormality of AKAP82mRNA results in the abnormality of AKAP82 expression.
方法:采用计算机辅助精液分析系统检测59例弱精子症患者(实验组)和28例健康成年男性(对照组)精液中精子的运动参数,用ELISA方法检测实验组患者血清抗精子抗体(AsAb)。使用低渗肿胀(HOS)试验对两组精子进行分析,通过RT-PCR方法对两组精子中AKAP82mRNA进行检测,实验组患者经提高精子活力活率、抗体专阴治疗后再用RT-PCR方法对其精子AKAP82mRNA进行检测和分析。
结果:①59例实验组(17例AsAb阳性,42例AsAb阴性)中有15例标本在RT-PCR检测中有发现有AKAP82mRNA表达缺失,28例对照组中发现2例有AKAP82mRNA表达缺失。有统计学意义(P＜0.05)。其中AsAb阳性组中有4例AKAP82mRNA表达缺失;AsAb阴性组中有11例AKAP82mRNA表达阴性,并无显著性差异(P＞0.05)。②低渗肿胀(HOS)试验中实验组精子尾部总肿胀率,g型精子率均低于对照组(P＜0.05);实验组中15例RT-PCR检测AKAP82mRNA缺失标本精子尾部总肿胀率,g型精子率低于实验组中AKAP82mRNA未缺失标本(P＜0.05)。③15例AKAP82mRNA缺失的患者经提高精子活力活率、抗体专阴治疗后RT-PCR方法检测发现AKAP82mRNA仍然缺失。
结论:弱精子症患者AKAP82mRNA表达缺失高于健康成年男性;AsAb阳性患者与阴性组AKAP82mRNA表达无显著差异;弱精子症患者低渗肿胀(HOS)试验精子尾部总肿胀率g型精子率低于健康成年男性;在弱精子症患者中AKAP82mRNA表达缺失患者HOS精子尾部总肿胀率g型精子率低于AKAP82mRNA表达阳性患者。
部分弱精子症患者精子的AKAP82异常可能是精子活力低下的原因之一。AKAP82mRNA缺失是造成AKAP82异常表达的原因。
Objective:AKAP82(A-kinase anchor proteins82) is one of A-kinase anchor proteins in sperm.In order to know the AKAP82mRNA levels of the patients with asthenospermia,we examinated the expressions of AKAP82mRNA in the patients and the healthy fertile men.
Methods:Sperm samples were obtained from 59 patients with asthenospermia (experimental group),and the normal sperms were obtained from 28 healthy fertile men(control group).And we estimated motility parameters with a computer-assisted semen analysis(CASA) technique.AsAb(antisperm antibody) of blood serum in the experimental group's was measured by ELISA(enzyme-linked immunosorbentassay). We measured the two groups AKAP82mRNA by reverse transcriptase PCR.At last, after the experimental group was treated,we measured the levels of AKAP82mRNA by reverse transcriptase PCR.
Results:Among the 59 patients,15 patients had been found with AKAP82mRNA microdeletion,and among the 28 healthy fertile men,2 had been found with AKAP82mRNA microdeletion,the percentage of g type sperm and the total tail swelling rate were significantly different(P＜0.05) between control group and experimental group.
Conclusion:Abnormality of AKAP82 expression in sperm may be one of the etiological factors in asthenospermia.Abnormality of AKAP82mRNA results in the abnormality of AKAP82 expression.
引文
[1]郭应禄,胡礼泉.男科学.北京:人民卫生出版社,2004,11-264.
[2]郭应禄,胡礼泉.男科学.第一版.北京:人民卫生出版社,2004,93
[3]The ESHRE Cap ri Workshop Group.Male infertility update.Hum Rep rod,1998,13(7):2025-2032.
[4]郭应禄,李宏军.男性不育症.北京:人民军医出版社,2003,36-106
[5]World Health Organization.The global burden of reproductive health[J].Progress in human reproduction research,1997,42:2-3.
[6]Niederberger C.The clinical conundrum of complete asthenospermia[J].J Androl,2004,25(5):679-680.
[7]曾春花,熊承良,官黄涛,等.少、弱精子症患者精浆、精子和血清中锌及性激素水平的相关分析.中国计划生育学杂志2007;15(6):354-358
[8]李畅,刘继红.舣厂酰环磷酸腺苷埘人精子运动功能的影响[J].中华男科学,2002,8(6):398-400.
[9]Johnson LR,Foster JA,Haig-Lade wig L,et al.Assembly of AKAP82,a protein kinase A anchor protein,into the fibrous heath of mouse sperm[J].Dev Bio(?),1997,192(2):340-350.
[10]Guraya SS Cellular and molecular biology of eapaeitation and acrosonle reaction in spermatozoa[J].(?)nt Rev Cytol,2000,199:1.64
[11]李畅,刘继红.弱精子症患者精子 A 激酶锚定蛋白 AKAP82异常表达的研究.中华男科学,2005,12(11):907-910.
[12]郭应禄,胡礼泉.男科学[M].北京:人民卫生出版社,2004,11-264;253-254.
[13]熊承良,吴明章,刘继红,等.人类精子学.武汉:湖北科学技术出版社,2002.133-137
[14]刘永杰,黄宇烽,邵永.复方玄驹胶囊改善精液质量的临床观察[J].中华男科杂志,2007,13(4):364-366.
[15]高章圈,张婉萍,彭玉梅.玄驹口服液治疗少、弱精子症的临床研究[J].中国计划生育学杂志,2006,124(2):103-105.
[16]李勇.浅谈五子衍宗丸治疗男子不育症[J].河南中医学院学报,2004,19(2):48-49.
[17]Carrera A,Gerton GL,Moss SB.The major fibrous sheathpolypeptide of mouse sperm:structural and functional similarities to the A-kinase anchoring proteins[J].Dev Biol,1994,165(1):272-284.
[18]Carrera A,Gerton GL,Moss SB.The major fibrous sheathpolypeptide of mouse sperm:structural and functional similarities to the A-kinase anchoring proteins[J].Dev Biol,1994,165(1):272-284.
[19]Johnson LR,Foster JA,Haig-Ladewig L,et al.Assembly of AKAP82,a protein kinase A anchor protein,into the fibrous sheath of mouse sperm[J].Dev Biol,1997,192(2):340-350.
[20]Turner RM,Eriksson RL,Gerton GL,et al.Relationship between sperm motility and the processing and tyrosine phosphorylation of two human sperm fibrous sheath proteins,prohAKAP82 and hAKAP82[J].Mol Hum Reprod,1999,5(9):816-824.
[21]Turner RM,Foster JA,Gerton GL,et al.Molecular evaluation of two major human sperm fibrous sheath proteins,prohAKAP82 and hAKAP82,in stump tail sperm[J].Fertil Steril,2001,76(2):267-274.
[22]熊承良,黄勋彬,沈继云,等.弱精子症患者精子中 ATP 和琥珀酸脱氢酶的含量与精子活率的关系.同济医科大学学报 1999;28(4):284-291
[1]金龙金,张春玲,楼哲丰,等.细胞生物学杂志 2007;29(4):573-575
[2]曾春花,熊承良,官黄涛,等.中国计划生育学杂志 2007;15(6):354-358
[3]熊承良,吴明章,刘继红,等.精子学.武汉:湖北科技出版社,2001:61-62,150-151,278-285
[4]金保方,杨晓玉,卞廷松,等.中华男科学杂志 2007;13(1):87-90
[5]Balercia G,Moretti S,Vignini A,et al.J Androl 2004;25(2):245-249
[6]Buffone MG,Calamera JC,Verstraeten SV,et al.Reproduction 2005;129(6):697-705
[7]Sanocka D,Miesel R,Jedrzejczak P,et al.J Androl1996;17(4):449-454
[8]Miesel R,Jedrzejczak P,Sanocka D,et al.Andrologia1997;29(2):77-83
[9]熊承良,黄勋彬,沈继云,等.同济医科大学学报 1999;28(4):284-291
[10]Matsushita K,Kitagawa K,Matsuyama T,et al.Brain Res 1996;743(1-2);362-365
[11]曾春花,熊承良,官黄涛,等.中国计划生育学杂志 2007;15(6):354-358
[12]Guraya SS.Cellular and molecular biology of capacitation and acrosome reaction in spermatozoa[J].Int Rev Cytol,2000,199(1):1-64.
[13]Carrera A,Gerton GL,Moss SB.The major fibrous sheath polypeptide of mouse sperm:structural and functional similarities to the Akinase anchoring proteins[J].Dev Biol,1994,165(1):272-284.
[14]Johnson LR,Foster JA,Haig-Ladewig L,et al.Assembly of AKAP82,a protein kinase A anchor protein,into the fibrous sheath of mousesperm[J].Dev Biol,1997,192(2):340-350.
[15]Turner RM,Johnson LR,Haig-Ladewig L,et al.An X-linked gene encodes a major human sperm fibrous sheath protein,hAKAP82.Genomic organization,protein kinase A-RII binding,and distribution of the precursor in the sperm tail[J].J Biol Chem,1998,273(48):32135-32141.
[16]Moss SB,Turner RM,Burkert KL,et al.Conservation and function of a bovine sperm A2kinase anchor protein homologous to mouse AKAP82[J].Biol Reprod,1999,61(2):335-342.
[17]J ha KN,Shivaji S.Identification of the major tyrosine phosphorrylated protein of capacitated hamster spermatozoa as a homologue of mammalian sperm a kinase anchoring protein[J].MolReprod Dev,2002,61 (2):258-270.
[18]Turner RM,Eriksson RL,Gerton GL,et al.Relationship between sperm motility and the processing and tyrosine phosphorylation of two human sperm fibrous sheath proteins,pro-hAKAP82 and hAKAP82[J].Mol Hum Reprod,1999,5 (9):816-824.
[19]Turner RM,Foster JA,Gerton GL,et al.Molecular evaluation of two major human sperm fibrous sheath proteins,pro-hAKAP82 and hAKAP82,in stump tail sperm[J].Fertil Steril,2001,76 (2):267-274.
[20]Lin RY,Moss SB,Rubin CS.Characterization of S-AKAP84,anovel developmentally regulated A kinase anchor protein of malegerm cells[J].J Biol Chem,1995,270 (46):27804-27811.
[21]Trendelenburg G,Hummel M,Riecken EO,et al.Molecular characterization of AKAP149,a novel A kinase anchor protein with a KH domain[J].Biochem Biophys Res Commun,1996,225 (1):313-319.
[22]Vijayaraghavan S,Liberty GA,Mohan J,et al.Isolation and molecular characterization of AKAPllO,a novel,sperm-specificprotein kinase A-anchoring protein[J].Mol Endocrinol,1999,13 (5):705-717.
[23]Feliciello A,Rubin CS,Avvedimento EV,et al.Expression of a kinase anchor protein 121 is regulated by hormones in thyroidand testicular germ cells[J].J Biol Chem,1998,273 (36):23361-23366.
[24]Cardone L,de Cristofaro T,Affaitati A,et al.A-kinase anchor protein 84/ 121 are targeted to mitochondria and mitotic spindles by overlapping aminoterminal motifs[J].J Mol Biol,2002,320 (3):663-675.
[25]Reinton N,Collas P,Haugen TB,et al.Localization of a novel human A-kinase-anchoring protein,hAKAP220,during spermatogenesis[J].Dev Biol,2000,223 (1):194-204.
[2]郭应禄,胡礼泉.男科学.第一版.北京:人民卫生出版社,2004,93
[3]The ESHRE Cap ri Workshop Group.Male infertility update.Hum Rep rod,1998,13(7):2025-2032.
[4]郭应禄,李宏军.男性不育症.北京:人民军医出版社,2003,36-106
[5]World Health Organization.The global burden of reproductive health[J].Progress in human reproduction research,1997,42:2-3.
[6]Niederberger C.The clinical conundrum of complete asthenospermia[J].J Androl,2004,25(5):679-680.
[7]曾春花,熊承良,官黄涛,等.少、弱精子症患者精浆、精子和血清中锌及性激素水平的相关分析.中国计划生育学杂志2007;15(6):354-358
[8]李畅,刘继红.舣厂酰环磷酸腺苷埘人精子运动功能的影响[J].中华男科学,2002,8(6):398-400.
[9]Johnson LR,Foster JA,Haig-Lade wig L,et al.Assembly of AKAP82,a protein kinase A anchor protein,into the fibrous heath of mouse sperm[J].Dev Bio(?),1997,192(2):340-350.
[10]Guraya SS Cellular and molecular biology of eapaeitation and acrosonle reaction in spermatozoa[J].(?)nt Rev Cytol,2000,199:1.64
[11]李畅,刘继红.弱精子症患者精子 A 激酶锚定蛋白 AKAP82异常表达的研究.中华男科学,2005,12(11):907-910.
[12]郭应禄,胡礼泉.男科学[M].北京:人民卫生出版社,2004,11-264;253-254.
[13]熊承良,吴明章,刘继红,等.人类精子学.武汉:湖北科学技术出版社,2002.133-137
[14]刘永杰,黄宇烽,邵永.复方玄驹胶囊改善精液质量的临床观察[J].中华男科杂志,2007,13(4):364-366.
[15]高章圈,张婉萍,彭玉梅.玄驹口服液治疗少、弱精子症的临床研究[J].中国计划生育学杂志,2006,124(2):103-105.
[16]李勇.浅谈五子衍宗丸治疗男子不育症[J].河南中医学院学报,2004,19(2):48-49.
[17]Carrera A,Gerton GL,Moss SB.The major fibrous sheathpolypeptide of mouse sperm:structural and functional similarities to the A-kinase anchoring proteins[J].Dev Biol,1994,165(1):272-284.
[18]Carrera A,Gerton GL,Moss SB.The major fibrous sheathpolypeptide of mouse sperm:structural and functional similarities to the A-kinase anchoring proteins[J].Dev Biol,1994,165(1):272-284.
[19]Johnson LR,Foster JA,Haig-Ladewig L,et al.Assembly of AKAP82,a protein kinase A anchor protein,into the fibrous sheath of mouse sperm[J].Dev Biol,1997,192(2):340-350.
[20]Turner RM,Eriksson RL,Gerton GL,et al.Relationship between sperm motility and the processing and tyrosine phosphorylation of two human sperm fibrous sheath proteins,prohAKAP82 and hAKAP82[J].Mol Hum Reprod,1999,5(9):816-824.
[21]Turner RM,Foster JA,Gerton GL,et al.Molecular evaluation of two major human sperm fibrous sheath proteins,prohAKAP82 and hAKAP82,in stump tail sperm[J].Fertil Steril,2001,76(2):267-274.
[22]熊承良,黄勋彬,沈继云,等.弱精子症患者精子中 ATP 和琥珀酸脱氢酶的含量与精子活率的关系.同济医科大学学报 1999;28(4):284-291
[1]金龙金,张春玲,楼哲丰,等.细胞生物学杂志 2007;29(4):573-575
[2]曾春花,熊承良,官黄涛,等.中国计划生育学杂志 2007;15(6):354-358
[3]熊承良,吴明章,刘继红,等.精子学.武汉:湖北科技出版社,2001:61-62,150-151,278-285
[4]金保方,杨晓玉,卞廷松,等.中华男科学杂志 2007;13(1):87-90
[5]Balercia G,Moretti S,Vignini A,et al.J Androl 2004;25(2):245-249
[6]Buffone MG,Calamera JC,Verstraeten SV,et al.Reproduction 2005;129(6):697-705
[7]Sanocka D,Miesel R,Jedrzejczak P,et al.J Androl1996;17(4):449-454
[8]Miesel R,Jedrzejczak P,Sanocka D,et al.Andrologia1997;29(2):77-83
[9]熊承良,黄勋彬,沈继云,等.同济医科大学学报 1999;28(4):284-291
[10]Matsushita K,Kitagawa K,Matsuyama T,et al.Brain Res 1996;743(1-2);362-365
[11]曾春花,熊承良,官黄涛,等.中国计划生育学杂志 2007;15(6):354-358
[12]Guraya SS.Cellular and molecular biology of capacitation and acrosome reaction in spermatozoa[J].Int Rev Cytol,2000,199(1):1-64.
[13]Carrera A,Gerton GL,Moss SB.The major fibrous sheath polypeptide of mouse sperm:structural and functional similarities to the Akinase anchoring proteins[J].Dev Biol,1994,165(1):272-284.
[14]Johnson LR,Foster JA,Haig-Ladewig L,et al.Assembly of AKAP82,a protein kinase A anchor protein,into the fibrous sheath of mousesperm[J].Dev Biol,1997,192(2):340-350.
[15]Turner RM,Johnson LR,Haig-Ladewig L,et al.An X-linked gene encodes a major human sperm fibrous sheath protein,hAKAP82.Genomic organization,protein kinase A-RII binding,and distribution of the precursor in the sperm tail[J].J Biol Chem,1998,273(48):32135-32141.
[16]Moss SB,Turner RM,Burkert KL,et al.Conservation and function of a bovine sperm A2kinase anchor protein homologous to mouse AKAP82[J].Biol Reprod,1999,61(2):335-342.
[17]J ha KN,Shivaji S.Identification of the major tyrosine phosphorrylated protein of capacitated hamster spermatozoa as a homologue of mammalian sperm a kinase anchoring protein[J].MolReprod Dev,2002,61 (2):258-270.
[18]Turner RM,Eriksson RL,Gerton GL,et al.Relationship between sperm motility and the processing and tyrosine phosphorylation of two human sperm fibrous sheath proteins,pro-hAKAP82 and hAKAP82[J].Mol Hum Reprod,1999,5 (9):816-824.
[19]Turner RM,Foster JA,Gerton GL,et al.Molecular evaluation of two major human sperm fibrous sheath proteins,pro-hAKAP82 and hAKAP82,in stump tail sperm[J].Fertil Steril,2001,76 (2):267-274.
[20]Lin RY,Moss SB,Rubin CS.Characterization of S-AKAP84,anovel developmentally regulated A kinase anchor protein of malegerm cells[J].J Biol Chem,1995,270 (46):27804-27811.
[21]Trendelenburg G,Hummel M,Riecken EO,et al.Molecular characterization of AKAP149,a novel A kinase anchor protein with a KH domain[J].Biochem Biophys Res Commun,1996,225 (1):313-319.
[22]Vijayaraghavan S,Liberty GA,Mohan J,et al.Isolation and molecular characterization of AKAPllO,a novel,sperm-specificprotein kinase A-anchoring protein[J].Mol Endocrinol,1999,13 (5):705-717.
[23]Feliciello A,Rubin CS,Avvedimento EV,et al.Expression of a kinase anchor protein 121 is regulated by hormones in thyroidand testicular germ cells[J].J Biol Chem,1998,273 (36):23361-23366.
[24]Cardone L,de Cristofaro T,Affaitati A,et al.A-kinase anchor protein 84/ 121 are targeted to mitochondria and mitotic spindles by overlapping aminoterminal motifs[J].J Mol Biol,2002,320 (3):663-675.
[25]Reinton N,Collas P,Haugen TB,et al.Localization of a novel human A-kinase-anchoring protein,hAKAP220,during spermatogenesis[J].Dev Biol,2000,223 (1):194-204.