安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2013年
11期
1387-1390
,共4页
周浩%董云华%孙杨%李红玲%沈涛%严新民
週浩%董雲華%孫楊%李紅玲%瀋濤%嚴新民
주호%동운화%손양%리홍령%침도%엄신민
不育%精液RNA%实时定量PCR%Kif2a
不育%精液RNA%實時定量PCR%Kif2a
불육%정액RNA%실시정량PCR%Kif2a
infertile%sperm RNA%real-time quantitative PCR%Kif2a
目的检测驱动蛋白重链成员2A(Kif2a)在男性精液中的表达,探讨Kif2a在不育男性精液中表达的意义。方法收集72例新鲜精液样本液化,经过离心等预处理后用TRIzol试剂提取精液RNA并逆转录成cDNA。运用实时荧光定量PCR检测精液中总Kif2a mRNA的量,并选取管家基因甘油醛-3-磷酸脱氢酶(GAPDH)作为内参对照。样本依据精液临床检测指标分为异常组和正常组。结果实时定量PCR法检测所得结果显示Kif2a mRNA存在于人类精液中。对Kif2a mRNA进行相对定量分析显示,在精子存活率、精子密度、精子活动率(a+b+c级精子)方面,异常组中 Kif2a mRNA的表达量较正常组明显降低,差异有统计学意义(P<0.05)。结论结合Kif2a在男性异常和正常精液中的差异性表达,其在精子的生成和受孕方面是否存在潜在的影响值得进一步探索。
目的檢測驅動蛋白重鏈成員2A(Kif2a)在男性精液中的錶達,探討Kif2a在不育男性精液中錶達的意義。方法收集72例新鮮精液樣本液化,經過離心等預處理後用TRIzol試劑提取精液RNA併逆轉錄成cDNA。運用實時熒光定量PCR檢測精液中總Kif2a mRNA的量,併選取管傢基因甘油醛-3-燐痠脫氫酶(GAPDH)作為內參對照。樣本依據精液臨床檢測指標分為異常組和正常組。結果實時定量PCR法檢測所得結果顯示Kif2a mRNA存在于人類精液中。對Kif2a mRNA進行相對定量分析顯示,在精子存活率、精子密度、精子活動率(a+b+c級精子)方麵,異常組中 Kif2a mRNA的錶達量較正常組明顯降低,差異有統計學意義(P<0.05)。結論結閤Kif2a在男性異常和正常精液中的差異性錶達,其在精子的生成和受孕方麵是否存在潛在的影響值得進一步探索。
목적검측구동단백중련성원2A(Kif2a)재남성정액중적표체,탐토Kif2a재불육남성정액중표체적의의。방법수집72례신선정액양본액화,경과리심등예처리후용TRIzol시제제취정액RNA병역전록성cDNA。운용실시형광정량PCR검측정액중총Kif2a mRNA적량,병선취관가기인감유철-3-린산탈경매(GAPDH)작위내삼대조。양본의거정액림상검측지표분위이상조화정상조。결과실시정량PCR법검측소득결과현시Kif2a mRNA존재우인류정액중。대Kif2a mRNA진행상대정량분석현시,재정자존활솔、정자밀도、정자활동솔(a+b+c급정자)방면,이상조중 Kif2a mRNA적표체량교정상조명현강저,차이유통계학의의(P<0.05)。결론결합Kif2a재남성이상화정상정액중적차이성표체,기재정자적생성화수잉방면시부존재잠재적영향치득진일보탐색。
Objective To detect the expression of kinesin heavy chain member 2A (Kif2a) in human ejaculate and discuss the significance of Kif2a in ejaculate of infertile man. Methods Human ejaculate samples were obtained from 72 subjects, sperm RNA was extracted from ejaculate cells by using TRIzol reagent after liquefaction and cen-trifugation, and the RNA was used for reverse transcription into cDNA. The amount of Kif2a mRNA was quantified by utilizing real-time quantitative PCR (RT-PCR);Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was se-lected as housekeeping gene. The subjects were grouped as abnormal and normal based on the clinical diagnosis of the ejaculate. Results The results of RT-PCR showed that the Kif2a mRNA is present in human ejaculates. Con-junction with clinical data of sperm vitality, concentration and total motility (progressive+nonprogressive), the ex-pression of Kif2a mRNA in abnormal groups was lower than that in normal groups, with the difference between them being statistically significant(P<0.05). Conclusion According to the distinct expression of Kif2a mRNA in ab-normal and normal ejaculate, whether the difference has potential impact on spermatogenesis and fertilization should be studied further.