精索静脉曲张大鼠睾丸iNOS的表达及生精冲剂对其表达影响的实验研究
摘要
目的：探讨精索静脉曲张大鼠睾丸中iNOS的表达及生精冲剂对精索静脉曲张大鼠iNOS表达的影响，从分子水平探讨及揭示精索静脉曲张不育的机制及生精冲剂治疗精索静脉曲张不育的作用机理。方法：选择雄性wistar大鼠(310+10g)60只，按时津和彦改良法制成精索静脉曲张大鼠模型，随机分为对照组20只(假手术组SOG)、模型组20只(精索静脉曲张VG)、治疗组20只(造模+中药VMG)。治疗组术后一周给生精冲剂(1.2g／300g)每日一次灌胃；模型组术后一周，给等剂量蒸馏水，每日一次灌胃。各组普食喂养三个月后，手术取睾丸组织观测睾丸重量、组织形态及iNOS表达。
结果：(1)VG组左睾重量明显低于SOG组(p＜0.001)，VMG组左睾重量高于VG组(p＜0.001)，VMG组与SOG组无明显差异。(2)VMG组病理组织学改变明显轻于VG组。(3)VG组睾丸组织iNOS表达量较SOG组与VMG组明显增强，VMG与SOG组间iNOS表达量稍有差异。结论：精索静脉曲张大鼠睾丸iNOS表达增强，生精冲剂对精索静脉曲张大鼠睾丸组织具有保护作用、能减少睾丸iNOS的表达。
Objective: To prove iNOS expression in varicocele rats testis.To discuss influence of Sengjing-soluable-granules to testicular iNOS expression in varicocele rats.Then discuss and reveal molecular level. M ethods:Experimental varicocele was created in male rats by partial ligation of left renal vein .The study were divided into 3 groups randomly:varicocele group(VG),shan operation group(SOG), treatment group(varicocele+chinese traditional medicine group VMG).The histology was observed at the 3rd postoperaative month.
Results:Et was found that(l) the left testicular weight in VG was far lower than that in SOG(p<0.001).(2) the pathologic changes in VG were severer than that in VMG.(3)Amount of iNOS positive leydig cell decreased in varicocele testicular tissue on the whole .most of leydig cell showed iNOS intensely negative in VMG rats testis. Conclusions: According to the experiment confirmed that soluble granules of Shengjing protected can protect the testicular tissues of varicocele rats and alleviate testicular pathologic damage ; The amount of iNOS expression decreased in varicocele rats testis.
结果：(1)VG组左睾重量明显低于SOG组(p＜0.001)，VMG组左睾重量高于VG组(p＜0.001)，VMG组与SOG组无明显差异。(2)VMG组病理组织学改变明显轻于VG组。(3)VG组睾丸组织iNOS表达量较SOG组与VMG组明显增强，VMG与SOG组间iNOS表达量稍有差异。结论：精索静脉曲张大鼠睾丸iNOS表达增强，生精冲剂对精索静脉曲张大鼠睾丸组织具有保护作用、能减少睾丸iNOS的表达。
Objective: To prove iNOS expression in varicocele rats testis.To discuss influence of Sengjing-soluable-granules to testicular iNOS expression in varicocele rats.Then discuss and reveal molecular level. M ethods:Experimental varicocele was created in male rats by partial ligation of left renal vein .The study were divided into 3 groups randomly:varicocele group(VG),shan operation group(SOG), treatment group(varicocele+chinese traditional medicine group VMG).The histology was observed at the 3rd postoperaative month.
Results:Et was found that(l) the left testicular weight in VG was far lower than that in SOG(p<0.001).(2) the pathologic changes in VG were severer than that in VMG.(3)Amount of iNOS positive leydig cell decreased in varicocele testicular tissue on the whole .most of leydig cell showed iNOS intensely negative in VMG rats testis. Conclusions: According to the experiment confirmed that soluble granules of Shengjing protected can protect the testicular tissues of varicocele rats and alleviate testicular pathologic damage ; The amount of iNOS expression decreased in varicocele rats testis.
引文
[1]Hwnang Choi,et al.The effect of experimental uaricocele oj thetestes of adolescant rats.Jrol 112(8):499 1990
[2]吴在德主编，外科学(第五版)2002．1．773
[3]徐开宏，等．精索静脉曲张研究进展．临床泌尿外科杂志1995．10(3)：184-187
[4]邱建宏，等．精索静脉曲张与男性生殖功能．男性学杂志1997，11(3)：176-177
[5]曾进，等．精索静脉曲张不育症患者睾丸的病理改变及机理探讨，男性学杂志，1993；7(1)：187-190
[6]Takihara H,Sakatoku J,Cocktt ATK.The pathophysi01ogy of varixoxele in male infertileity.Feritil steril 1991;55:861
[7]袁咏裳主编，临床泌尿外科诊疗手册，2000．3．1 35
[8]岩成浩，等．精索静脉曲张患者局部静脉血气分析，中华泌尿外科杂志，1989；27(1)：37
[9]Chakra borty,J,Hilim A P and jhuni hunwala,J S stagnation of blood in the rmcrocieoulatory usesels in the testes of men with varicocele j Androl 6:117 1985
[10]Nakamura M,Hall P Eet al.The mechanism by uhich body temperative inhibits protein biosynthesis in speronatids of rat testes.JBiol chem. 1980;255:2907
[11]Ross,J [12]Maclod J:Seminol ctytology in the presence oj varicoocele Fertil steril 1996:16:753
[13]Mohamed M,Wishahi,Anatomy oj the spermatic venous plexuss (pamainiform plexuss)in men with and uithout varocele interaoperative venographic study J urol 1992;147:1285
[14]王全兵，等．精索静脉曲张患者肾上腺代谢返流的研究，生殖与避孕药
1993；13(5)：396397
[15]蔡晓晴等．五羟色胺对精索静脉曲张患者睾丸生精功能的影响，中华泌尿外科杂志，195；16：1
[16]李汛，等．精索静脉旁路术肾素-血管紧张素测定研究，中华纤维外科杂志，1995；8(18)：3
[17]Wang R, Kiling Varicocde in the rat:Anew, ecperimentalmodel Effect on histology ultrastucture and temperature of the testis and epididymis Vrol Res1991;22(41):276
[18]Mc Cluere RD,Crawford VA,marrero ra et al subdinval varicocele:the effectiveness oj varicoceletomy J Urol 1991;145:789
[19]姜永光，等．精索静脉曲张不育患者精浆．糖甙酶的检测意义，临床泌尿外科杂志，1995；10(1)20-22
[20]姜永光，等．精索静脉曲张不育患者精子核蛋白，精子膜麦芽凝集素受体的检测和意义，中华尿外科杂志，1994；12：15(6)456-457
[21]Golomb J,vardinon N,Homonnai ZT et al.Ddemonstration of antispermatozoal antibodies in baricocele-dated infertility with an enzyme-linked.immunosorbentassay(elisa)fertile steril, 1986;45(3):397
[22]Nagao k r et al.lomparison oj gondel function between fertile and infertile men with variocdes fertile steril, 1998;49(2): 199
[23]庄乾元．精索静脉曲张不育患者血清生殖激素水平分析，同济医科大学，1993；22(41)：276
[24]Mitopoulos-D Delilonstantions-G;Nitric oxide synthase and xanthine oxidese activities in the spermatic vein of patients with varicolcdes a potential role for notric oxide and peroxynitite in sperm dysfunetions.J uol 156(6):1952-8' 1996
[25]息金波，等。精索静脉曲张大鼠睾丸LPO与SOD含量及ACE活力的变化。生殖与避孕。1992 12(4)63-67
[26]Plante M.Lanirande E and Gagnon c.Reactive exygen species released by activated neutroplnle,but not by defficent spermatozoa are sufficient to affect normal sperm notility fertile steril 1994;62:387
[27]Krauz,G,Mills C,rogers s,Tan land Aitken R.et al.J Stimulation of oxidation gen?ration by human sperm suspend sions using phorbol esters and formy peptides relationship with motility and fertitizaion in c=vito.Futil steril 1994;62:599
[28]Mancini-A.et al.Relatinship between sperm cell ubiquinonevand seminal parameters in subjects wirh and uithoust varicocele.Andrologia 1998 FebMar:1998;30(1);1-4
[29]Jones R,mann jand sherins R.et al.J peroxidative break down of phospholimpids in uman spermatozoa spermicidal propertyes of faffy acid peroxides and protective action of seminal plasma.Fertil steril.1979;31:531
[30]Aitken Rlrvine D,and wu F.C.et al.prospective analysis of spermocyte fustion and reactive oxygen species generation as criteria for te diagnosis of infertileity.Amer.jobst gynec 1991; 164:542
[31]Alvarez JG,touchstone JG.et al.Spontaneous lipid peroxidation and production of hyerogen peroxide and superoxide in human spermatozoa superoxide dismutase as major inzyme protectant against oxygen toxitoity.J.Androl. 1987;8:338
[32]Lenzi A,picardo M.et al.Glutathione treatment of duspermia effect on the lipoperoxidation prodess Hum reprod. 1994;9:2044
[33]Weinberg J,bdoty E.et al.Nitricoxide inhibition of human sperm motility.Fertil Steril. 1995;64:408
[34]姜永光等。精索静脉曲张不育患者精子核蛋白精子膜麦芽凝集素受体的检测和意义，中华泌尿外科杂志，1994；12：15(6)456-457
[35]钟慈声，孙安阳 一氧化氮的生物医学[M]上海：上海医科大学出版社，1997
[36]Burnett AL,Maguire MP,Chamness,Sletal Characterizationand localization of nitricoxidesynthase in the human prostate[J]Urology,1995,45(3):435～439
[37]Syder SH.Nature,1995;377:196
[38]ZiniA,BryanMK, Magid Ms et al.Immunohistoche mical localization of
endothelial nitric oxide synthase in human testis,epididymis,and vas deferens suggests a possible role for nitric oxid in spermatogenesis,sperm maturation and programmed cell death[J] Biol Reprod,1996,55(5):935-941
[39]Sabanegh E JR,Dewire D.Fertil Steril 1994;S9
[40]田溪泉，潘天明，郭应禄，一氧化氮对精子运动功能的影响，中华泌尿外科杂志，1999，20(8)：496
[41]Snyder SH,Bredt Ds.Biological roles of nitric oxide.scientific Americian, 1992,226:28
[42]Rossell;M,Dubey Rk,Imthurn B etal Effects of nitric odide on human spermatozoa :evidence that nitric oxide decreases sperm motility and induces sperm toxicity[J] Hum Reprod,1995,10(7):1786-1790
[43]Noel AA,Fallek SR,Hobson Rw,et al.Inhibition of nitric oxide synthase attenuates primed microvascular permeability in the in vivo microcirculation[J].J Vasc Surg, 1995,22(6):661-670
[44]Ochsendorf Fr,Thiele J, Fuchs J et al.Chemiluminescence in semen of infertile men [J]Andrologia,1994,26(5):289-293
[45]陈国武，郑松，吴裕伦等。精液白细胞与精子功能间关系的研究[J]男性学杂志，1998，12(2)：82-85
[46]Selley ML,Lacey MJ,Bartlett MR et al,Content of significant amounts of a cytotoxic end -product of lipid peroxidation in human semen [J]J Reprod Fertil, 1991,92(1):291-298
[47]郑荣梁．自由基对人精子功能的调控[J]，中国工业医学杂志，1999，12(2)：115
[48]Yeoman RR,Jones wD,Rizk BM,Evidence for nitric oxide regulation of hamster sperm hyperactivation [J]J Androl, 1998,19(1):58-64
[49]Yeoman RR,Hazelton JM,,Aksel s et al.Inhibition of nitric of nitric oxide (no)differentially effects hyperactivation and acrosome reaction in live hamster sperm[J],Fertil, 1994,S10
[50]安崇臣，余朋干主编。中医男科症治备要。科学技术出版社。1992；8
[51]陈晓平，等。中医药治疗精索静脉曲张94例。新中医。1998；8：27-28
[52]戚广崇等。通精煎治疗精索静脉曲张不育症102例临床观察。1988：8(10)626-627
[53]安立文，等。手术配合中药治疗精索静脉曲张男性不育症。中医药学报。1992：1：33
[54]安立文，等。生精冲剂对精索静脉曲张不育症患者术后精子形态学的影响。中医杂志，1998：39(6)：349-351
[55]夏明歧，等。中药内服外洗治疗精索静脉曲张[J]．四川中医，1999，17(7)：41．
[56]Rageth JC Long-tem fesults of Varicocelectomy Urol int 1992；48(3)：327
[57]薄焦杰，等。精索内静脉高位结扎术治疗精索静脉曲张不育症随机对照研究，男性学杂志，1996：10：3：141-145
[58]吴宏飞，等。精索筋膜肌管折叠术治疗精索静脉曲张及远期随访，中华泌尿外科杂志，1995；1(16)：135-137
[59]黄红卫，等。腹壁下静脉转流术治疗精索静脉曲张，江西医学院学报2000；40(1)：110～111
[60]赵斌，傅毅敏，刘桂兰．当归注射液治疗精索静脉曲张30例[J]．针灸临床杂志，1999，15(2)：36．
[61]W．布卢姆D．W．福西特主编组织学。科学出版社，1984。5：787-801
[62]AitkenRJ,BuckinghamD,Harkiss,Detal Use of axanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa[J] JReprod Fertil, 1993,97(2):441～450
[63]D Agatar,Vicarie,Moncadamletal Generation of reactive oxygen species in subguoups of in fertilemen[J] Intj And rol, 1990,13(5):344～351
[64]Iwasakia,Gagnonc Formation of reactive oxygen species in sper matozoaof in fertil epatients[J]Fertil Steril, 1992,57(2):409～416
[65]Delamirande,Gagnonc Reactive oxygen species and human spermatozoa Ⅱ Depletion of adenoslnetrlphosphate plays an important role in the inhibition of spermmotility[J]Jandrol, 1992,13(5):379～386
[66]Aitken RJ,Harkissd,Buckingham Detal Relationship between iron
catalysed lipid peroxidation potential and human sperm function[J] JReprodFertil, 1993,98(1):257～265
[67]Huszarg,Voguel Correlation between the rate of lipid peroxidation and cell ularmaturity as measured by creat inekinase activity in human spermatozoa[J]Jandrol, 1994,15(1):71～77
[68]OchsendorfFR,ThieleJ,Fuchs Jetal Chemiluminescenceinse men of infertile men[J]And rologia, 1994,26(5):289～29
[69]陈国武，郑松，吴裕伦等．精液白细胞与精子功能间关系的研究[J]男性学杂志，1998，12(2)：82～85
[70]Selley.ML,laceyMJ,Bartlerr.MRetal.Contento fsignificantamounts of acytotoxicend product of lipid peroxidation in humanse men[J]JReprodFertil,1991,92(1):291～298
[71] 郑荣梁 自由基对人精子功能的调控[J]中国工业医学杂志，1999，12(2)：115
[72]ZhangH,zheng RL possible role of nitricoxide on fertile and as the nozoo spermic in fertile human sperm function[J]Free radio Res, 1996,25(4):347～353
[73]LewisSE,DonnellyET,Sterl ingesetal Nitricoxide synthase and nitrite production in human spermatozoa,evidence that end ogenous nitricoxide is beneficial to sperm motility[J]MolHumRerod, 1996,2(11):873～878
[74]Grisham MB Interaction between nitricoxide and super oxide:Role in modulating leukocy teadhesion in the postischemicmicrovasculature[J]TransplantProc 1995,27(5):2842～2843
[75] 凌亦凌，黄善生，谷振勇过氧亚硝基阴离子的细胞代谢及病理损伤作用[J]生理科学进展，1999，30(1)：71～73．
[76]Mitropoluos. D, DeliconstantinosG, Zervas. Aedal. Nitricoxidesynth aseandxanthine oxidase activitiesinthe spermaticvein of patients with varicoceleapodential role for nitricoxideand peroxynitritein sperm dysfunction[J] JUrol, 1996,156(6):1952～1958
[77]李育浩，等。中药药理与临床，1992；8(3)：4-
[2]吴在德主编，外科学(第五版)2002．1．773
[3]徐开宏，等．精索静脉曲张研究进展．临床泌尿外科杂志1995．10(3)：184-187
[4]邱建宏，等．精索静脉曲张与男性生殖功能．男性学杂志1997，11(3)：176-177
[5]曾进，等．精索静脉曲张不育症患者睾丸的病理改变及机理探讨，男性学杂志，1993；7(1)：187-190
[6]Takihara H,Sakatoku J,Cocktt ATK.The pathophysi01ogy of varixoxele in male infertileity.Feritil steril 1991;55:861
[7]袁咏裳主编，临床泌尿外科诊疗手册，2000．3．1 35
[8]岩成浩，等．精索静脉曲张患者局部静脉血气分析，中华泌尿外科杂志，1989；27(1)：37
[9]Chakra borty,J,Hilim A P and jhuni hunwala,J S stagnation of blood in the rmcrocieoulatory usesels in the testes of men with varicocele j Androl 6:117 1985
[10]Nakamura M,Hall P Eet al.The mechanism by uhich body temperative inhibits protein biosynthesis in speronatids of rat testes.JBiol chem. 1980;255:2907
[11]Ross,J
[13]Mohamed M,Wishahi,Anatomy oj the spermatic venous plexuss (pamainiform plexuss)in men with and uithout varocele interaoperative venographic study J urol 1992;147:1285
[14]王全兵，等．精索静脉曲张患者肾上腺代谢返流的研究，生殖与避孕药
1993；13(5)：396397
[15]蔡晓晴等．五羟色胺对精索静脉曲张患者睾丸生精功能的影响，中华泌尿外科杂志，195；16：1
[16]李汛，等．精索静脉旁路术肾素-血管紧张素测定研究，中华纤维外科杂志，1995；8(18)：3
[17]Wang R, Kiling Varicocde in the rat:Anew, ecperimentalmodel Effect on histology ultrastucture and temperature of the testis and epididymis Vrol Res1991;22(41):276
[18]Mc Cluere RD,Crawford VA,marrero ra et al subdinval varicocele:the effectiveness oj varicoceletomy J Urol 1991;145:789
[19]姜永光，等．精索静脉曲张不育患者精浆．糖甙酶的检测意义，临床泌尿外科杂志，1995；10(1)20-22
[20]姜永光，等．精索静脉曲张不育患者精子核蛋白，精子膜麦芽凝集素受体的检测和意义，中华尿外科杂志，1994；12：15(6)456-457
[21]Golomb J,vardinon N,Homonnai ZT et al.Ddemonstration of antispermatozoal antibodies in baricocele-dated infertility with an enzyme-linked.immunosorbentassay(elisa)fertile steril, 1986;45(3):397
[22]Nagao k r et al.lomparison oj gondel function between fertile and infertile men with variocdes fertile steril, 1998;49(2): 199
[23]庄乾元．精索静脉曲张不育患者血清生殖激素水平分析，同济医科大学，1993；22(41)：276
[24]Mitopoulos-D Delilonstantions-G;Nitric oxide synthase and xanthine oxidese activities in the spermatic vein of patients with varicolcdes a potential role for notric oxide and peroxynitite in sperm dysfunetions.J uol 156(6):1952-8' 1996
[25]息金波，等。精索静脉曲张大鼠睾丸LPO与SOD含量及ACE活力的变化。生殖与避孕。1992 12(4)63-67
[26]Plante M.Lanirande E and Gagnon c.Reactive exygen species released by activated neutroplnle,but not by defficent spermatozoa are sufficient to affect normal sperm notility fertile steril 1994;62:387
[27]Krauz,G,Mills C,rogers s,Tan land Aitken R.et al.J Stimulation of oxidation gen?ration by human sperm suspend sions using phorbol esters and formy peptides relationship with motility and fertitizaion in c=vito.Futil steril 1994;62:599
[28]Mancini-A.et al.Relatinship between sperm cell ubiquinonevand seminal parameters in subjects wirh and uithoust varicocele.Andrologia 1998 FebMar:1998;30(1);1-4
[29]Jones R,mann jand sherins R.et al.J peroxidative break down of phospholimpids in uman spermatozoa spermicidal propertyes of faffy acid peroxides and protective action of seminal plasma.Fertil steril.1979;31:531
[30]Aitken Rlrvine D,and wu F.C.et al.prospective analysis of spermocyte fustion and reactive oxygen species generation as criteria for te diagnosis of infertileity.Amer.jobst gynec 1991; 164:542
[31]Alvarez JG,touchstone JG.et al.Spontaneous lipid peroxidation and production of hyerogen peroxide and superoxide in human spermatozoa superoxide dismutase as major inzyme protectant against oxygen toxitoity.J.Androl. 1987;8:338
[32]Lenzi A,picardo M.et al.Glutathione treatment of duspermia effect on the lipoperoxidation prodess Hum reprod. 1994;9:2044
[33]Weinberg J,bdoty E.et al.Nitricoxide inhibition of human sperm motility.Fertil Steril. 1995;64:408
[34]姜永光等。精索静脉曲张不育患者精子核蛋白精子膜麦芽凝集素受体的检测和意义，中华泌尿外科杂志，1994；12：15(6)456-457
[35]钟慈声，孙安阳 一氧化氮的生物医学[M]上海：上海医科大学出版社，1997
[36]Burnett AL,Maguire MP,Chamness,Sletal Characterizationand localization of nitricoxidesynthase in the human prostate[J]Urology,1995,45(3):435～439
[37]Syder SH.Nature,1995;377:196
[38]ZiniA,BryanMK, Magid Ms et al.Immunohistoche mical localization of
endothelial nitric oxide synthase in human testis,epididymis,and vas deferens suggests a possible role for nitric oxid in spermatogenesis,sperm maturation and programmed cell death[J] Biol Reprod,1996,55(5):935-941
[39]Sabanegh E JR,Dewire D.Fertil Steril 1994;S9
[40]田溪泉，潘天明，郭应禄，一氧化氮对精子运动功能的影响，中华泌尿外科杂志，1999，20(8)：496
[41]Snyder SH,Bredt Ds.Biological roles of nitric oxide.scientific Americian, 1992,226:28
[42]Rossell;M,Dubey Rk,Imthurn B etal Effects of nitric odide on human spermatozoa :evidence that nitric oxide decreases sperm motility and induces sperm toxicity[J] Hum Reprod,1995,10(7):1786-1790
[43]Noel AA,Fallek SR,Hobson Rw,et al.Inhibition of nitric oxide synthase attenuates primed microvascular permeability in the in vivo microcirculation[J].J Vasc Surg, 1995,22(6):661-670
[44]Ochsendorf Fr,Thiele J, Fuchs J et al.Chemiluminescence in semen of infertile men [J]Andrologia,1994,26(5):289-293
[45]陈国武，郑松，吴裕伦等。精液白细胞与精子功能间关系的研究[J]男性学杂志，1998，12(2)：82-85
[46]Selley ML,Lacey MJ,Bartlett MR et al,Content of significant amounts of a cytotoxic end -product of lipid peroxidation in human semen [J]J Reprod Fertil, 1991,92(1):291-298
[47]郑荣梁．自由基对人精子功能的调控[J]，中国工业医学杂志，1999，12(2)：115
[48]Yeoman RR,Jones wD,Rizk BM,Evidence for nitric oxide regulation of hamster sperm hyperactivation [J]J Androl, 1998,19(1):58-64
[49]Yeoman RR,Hazelton JM,,Aksel s et al.Inhibition of nitric of nitric oxide (no)differentially effects hyperactivation and acrosome reaction in live hamster sperm[J],Fertil, 1994,S10
[50]安崇臣，余朋干主编。中医男科症治备要。科学技术出版社。1992；8
[51]陈晓平，等。中医药治疗精索静脉曲张94例。新中医。1998；8：27-28
[52]戚广崇等。通精煎治疗精索静脉曲张不育症102例临床观察。1988：8(10)626-627
[53]安立文，等。手术配合中药治疗精索静脉曲张男性不育症。中医药学报。1992：1：33
[54]安立文，等。生精冲剂对精索静脉曲张不育症患者术后精子形态学的影响。中医杂志，1998：39(6)：349-351
[55]夏明歧，等。中药内服外洗治疗精索静脉曲张[J]．四川中医，1999，17(7)：41．
[56]Rageth JC Long-tem fesults of Varicocelectomy Urol int 1992；48(3)：327
[57]薄焦杰，等。精索内静脉高位结扎术治疗精索静脉曲张不育症随机对照研究，男性学杂志，1996：10：3：141-145
[58]吴宏飞，等。精索筋膜肌管折叠术治疗精索静脉曲张及远期随访，中华泌尿外科杂志，1995；1(16)：135-137
[59]黄红卫，等。腹壁下静脉转流术治疗精索静脉曲张，江西医学院学报2000；40(1)：110～111
[60]赵斌，傅毅敏，刘桂兰．当归注射液治疗精索静脉曲张30例[J]．针灸临床杂志，1999，15(2)：36．
[61]W．布卢姆D．W．福西特主编组织学。科学出版社，1984。5：787-801
[62]AitkenRJ,BuckinghamD,Harkiss,Detal Use of axanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa[J] JReprod Fertil, 1993,97(2):441～450
[63]D Agatar,Vicarie,Moncadamletal Generation of reactive oxygen species in subguoups of in fertilemen[J] Intj And rol, 1990,13(5):344～351
[64]Iwasakia,Gagnonc Formation of reactive oxygen species in sper matozoaof in fertil epatients[J]Fertil Steril, 1992,57(2):409～416
[65]Delamirande,Gagnonc Reactive oxygen species and human spermatozoa Ⅱ Depletion of adenoslnetrlphosphate plays an important role in the inhibition of spermmotility[J]Jandrol, 1992,13(5):379～386
[66]Aitken RJ,Harkissd,Buckingham Detal Relationship between iron
catalysed lipid peroxidation potential and human sperm function[J] JReprodFertil, 1993,98(1):257～265
[67]Huszarg,Voguel Correlation between the rate of lipid peroxidation and cell ularmaturity as measured by creat inekinase activity in human spermatozoa[J]Jandrol, 1994,15(1):71～77
[68]OchsendorfFR,ThieleJ,Fuchs Jetal Chemiluminescenceinse men of infertile men[J]And rologia, 1994,26(5):289～29
[69]陈国武，郑松，吴裕伦等．精液白细胞与精子功能间关系的研究[J]男性学杂志，1998，12(2)：82～85
[70]Selley.ML,laceyMJ,Bartlerr.MRetal.Contento fsignificantamounts of acytotoxicend product of lipid peroxidation in humanse men[J]JReprodFertil,1991,92(1):291～298
[71] 郑荣梁 自由基对人精子功能的调控[J]中国工业医学杂志，1999，12(2)：115
[72]ZhangH,zheng RL possible role of nitricoxide on fertile and as the nozoo spermic in fertile human sperm function[J]Free radio Res, 1996,25(4):347～353
[73]LewisSE,DonnellyET,Sterl ingesetal Nitricoxide synthase and nitrite production in human spermatozoa,evidence that end ogenous nitricoxide is beneficial to sperm motility[J]MolHumRerod, 1996,2(11):873～878
[74]Grisham MB Interaction between nitricoxide and super oxide:Role in modulating leukocy teadhesion in the postischemicmicrovasculature[J]TransplantProc 1995,27(5):2842～2843
[75] 凌亦凌，黄善生，谷振勇过氧亚硝基阴离子的细胞代谢及病理损伤作用[J]生理科学进展，1999，30(1)：71～73．
[76]Mitropoluos. D, DeliconstantinosG, Zervas. Aedal. Nitricoxidesynth aseandxanthine oxidase activitiesinthe spermaticvein of patients with varicoceleapodential role for nitricoxideand peroxynitritein sperm dysfunction[J] JUrol, 1996,156(6):1952～1958
[77]李育浩，等。中药药理与临床，1992；8(3)：4-