中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2010年
2期
362-367
,共6页
李传连%楼哲丰%黄学锋%吴永根%张李雅%吕建新%金龙金
李傳連%樓哲豐%黃學鋒%吳永根%張李雅%呂建新%金龍金
리전련%루철봉%황학봉%오영근%장리아%려건신%금룡금
弱精子症%基因%ND3%基因%ND4L%核苷酸变异%单体型
弱精子癥%基因%ND3%基因%ND4L%覈苷痠變異%單體型
약정자증%기인%ND3%기인%ND4L%핵감산변이%단체형
Asthenospermia%Genes%ND3%Genes%ND4L%Nucleotide variation%Haplogroup
目的:对弱精子症(AST) 精子线粒体DNA ND3、ND4L基因突变检测和分析,探索弱精子症致病的分子机制.方法:收集弱精子症患者50例,年龄匹配的对照42例,用密度梯度离心将弱精子症患者和对照组的不同活力精子进行分离,扩增线粒体ND3、ND4L基因,测序和比对,比较弱精子症组和对照组线粒体ND3、ND4L基因核苷酸变异和单体型差异.结果:在弱精子症组和对照组中共检测出22个变异位点,其中A10157G和A10313C未见报道,G10320A、A10398G、T10609C为错义突变.A10398G和C10400T核苷酸变异率在弱精子症组显著低于对照组(P<0.05),G10310A核苷酸变异率在弱精子症组显著高于对照组(P<0.05);单体型分析可见弱精子症组单体型N百分率(33/50)显著高于对照组(14/42)(P<0.05),单体型R9在弱精子症组(15/50)的比例显著高于对照组(4/42)(P<0.05);单体型F1、F2和R9的前向运动精子百分率显著低于单体型M和M rest(P<0.05).对弱精子症同一病例不同活力精子进行检测,有2例不同活力的精子的线粒体单体型存在差异,活力好的精子为单体型M,而活力差的精子为单体型N;在50例弱精子症中有2例活力中等和活力差的精子标本中检测出G10310A异质性突变,而在活力好的精子标本中无G10310A突变.结论:线粒体单体型与精子活力可能存在一定的相关性;线粒体DNA 10398G-10400T多态性可能是精子活力的有益因素,线粒体DNA G10310A突变可能是精子活力的有害因素.
目的:對弱精子癥(AST) 精子線粒體DNA ND3、ND4L基因突變檢測和分析,探索弱精子癥緻病的分子機製.方法:收集弱精子癥患者50例,年齡匹配的對照42例,用密度梯度離心將弱精子癥患者和對照組的不同活力精子進行分離,擴增線粒體ND3、ND4L基因,測序和比對,比較弱精子癥組和對照組線粒體ND3、ND4L基因覈苷痠變異和單體型差異.結果:在弱精子癥組和對照組中共檢測齣22箇變異位點,其中A10157G和A10313C未見報道,G10320A、A10398G、T10609C為錯義突變.A10398G和C10400T覈苷痠變異率在弱精子癥組顯著低于對照組(P<0.05),G10310A覈苷痠變異率在弱精子癥組顯著高于對照組(P<0.05);單體型分析可見弱精子癥組單體型N百分率(33/50)顯著高于對照組(14/42)(P<0.05),單體型R9在弱精子癥組(15/50)的比例顯著高于對照組(4/42)(P<0.05);單體型F1、F2和R9的前嚮運動精子百分率顯著低于單體型M和M rest(P<0.05).對弱精子癥同一病例不同活力精子進行檢測,有2例不同活力的精子的線粒體單體型存在差異,活力好的精子為單體型M,而活力差的精子為單體型N;在50例弱精子癥中有2例活力中等和活力差的精子標本中檢測齣G10310A異質性突變,而在活力好的精子標本中無G10310A突變.結論:線粒體單體型與精子活力可能存在一定的相關性;線粒體DNA 10398G-10400T多態性可能是精子活力的有益因素,線粒體DNA G10310A突變可能是精子活力的有害因素.
목적:대약정자증(AST) 정자선립체DNA ND3、ND4L기인돌변검측화분석,탐색약정자증치병적분자궤제.방법:수집약정자증환자50례,년령필배적대조42례,용밀도제도리심장약정자증환자화대조조적불동활력정자진행분리,확증선립체ND3、ND4L기인,측서화비대,비교약정자증조화대조조선립체ND3、ND4L기인핵감산변이화단체형차이.결과:재약정자증조화대조조중공검측출22개변이위점,기중A10157G화A10313C미견보도,G10320A、A10398G、T10609C위착의돌변.A10398G화C10400T핵감산변이솔재약정자증조현저저우대조조(P<0.05),G10310A핵감산변이솔재약정자증조현저고우대조조(P<0.05);단체형분석가견약정자증조단체형N백분솔(33/50)현저고우대조조(14/42)(P<0.05),단체형R9재약정자증조(15/50)적비례현저고우대조조(4/42)(P<0.05);단체형F1、F2화R9적전향운동정자백분솔현저저우단체형M화M rest(P<0.05).대약정자증동일병례불동활력정자진행검측,유2례불동활력적정자적선립체단체형존재차이,활력호적정자위단체형M,이활력차적정자위단체형N;재50례약정자증중유2례활력중등화활력차적정자표본중검측출G10310A이질성돌변,이재활력호적정자표본중무G10310A돌변.결론:선립체단체형여정자활력가능존재일정적상관성;선립체DNA 10398G-10400T다태성가능시정자활력적유익인소,선립체DNA G10310A돌변가능시정자활력적유해인소.
AIM: To explore the molecular mechanism of asthenospermia(AST) by preliminary screening of nucleotide sequences from the ND3 and ND4L genes of mitochondrial DNA(mtDNA). METHODS: Samples from 50 AST patients and 42 age-matched normal controls were collected according to the WHO criteria. Density gradient centrifugation was applied to separate spermatozoa with different vigor. The ND3 and ND4L genes of mtDNA were amplified and sequenced directly from the extracted genomic DNA from AST patients and normal controls. The sequences were compared with revised Cambridge Reference Sequence(rCRS) to analyze the variants. RESULTS: A total of 22 nucleotide variations were found in ND3 and ND4L genes of mtDNA in asthenospermia group and control group. G10320A, A10398G and T10609C were missense mutations, while A10157G and A10313C were the reported for the first time in this study. Haplotype N in patients with AST(33/50) was higher than that in control group(14/42, P<0.05), and haplotype R9 in patients with AST(15/50) was also higher than that in control group(4/42, P<0.05) through genetic testing of ND3 gene. Rates of sperm progressive motility of haplotype F1, F2 and R9 were significantly lower than those of haplotype M and M rest. Two haplotype differences, haplotype M and N, were found in the same AST patient's spermatozoas which had different vigor. Haplotype M had stronger vigor, while haplotype N had lower vigor. By sequencing ND3 gene of mtDNA from 50 AST patients, we detected G10310A heteroplasmic mutation in 2 specimens of asthenospermia with poor and moderate motility spermatozoa, respectively. No mutation occurred in good motility spermatozoa. CONCLUSION: Haplotype of mitochondrial may have some correlation with sperm motility. The nt10398G-10400T polymorphisms may have benefit for sperm motility, whereas the mutation in nt10310A may impair sperm motility.