Zn缺乏对人精子VDAC mRNA的影响
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国家自然科学基金面上项目(30872575, 30973199)


Effects of intracellular zinc depletion on the abundance of VDAC mRNA in cultured human spermatozoa
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    摘要:

    目的:探讨Zn离子缺乏对成熟精子活动能力和电压依赖性离子通道(voltage-dependent anion channel,VDAC)的mRNA丰度的影响-方法:从3个正常生育者捐献所得的精液经非连续Percoll梯度离心获得精子样本,并将其分3组连续培养24 h:对照组;加入Zn离子螯合剂的TPEN组(2 -滋mol/L);同时加入Zn离子螯合剂N,N,N',N'-tetrakis(2-pyridylmethy1) Ethylenediamine (TPEN)和Zn离子(5 -滋mol/L)组-行CASA和荧光定量PCR(qPCR)分别分析不同时间的精子活动能力和24 h后各VDAC亚型mRNA丰度-结果:CASA显示:培养3 h,TPEN组与对照组比较,精子活力明显下降(P < 0.05)-qPCR结果显示:培养24 h后,TPEN组VDAC3 mRNA丰度明显低于对照组(P < 0.05)-而同时加入TPEN和Zn离子组与对照组比较,差异无统计学意义-结论:Zn离子缺乏使VDAC3 mRNA丰度降低,可能影响到精子的正常功能-

    Abstract:

    Objective:To investigate whether intracellular zinc depletion can actually change spermatozoa motility and the abundance of voltage-dependent anion channel(VDAC) mRNA in cultured human spermatozoa. Methods: Spermatozoa were separated from ejaculates of three donors using a discontinuous Percoll gradient centrifugation. Each spermatozoa specimen were divided into 3 groups,one was exposed to a cell membrane-permeable zinc chelator N,N,N',N'-tetrakis(2-pyridylmethy1) Ethylenediamine (TPEN) (2 -滋mol/L) for 24 hours,one was to TPEN plus zinc sulfate(5 -滋mol/L) for 24 hours,and one was as control. CASA and quantitative real-time PCR(qPCR) were performed to detect sperm motility characteristics,mRNA abundance and difference of the three VDAC subtypes between three different treatment groups respectively. Results:The results of CASA demonstrated that exposure of spermatozoa to TPEN for three hours significantly decreased sperm motility compared with the control group(P = 0.049). qPCR demonstrated that VDAC3 mRNA level of TPEN group were significantly lower than that of control group,while no significant difference was found between TPEN plus zinc sulfate treatment group and control group. Co-addition of zinc partly reversed TPEN-induced alterations of sperm motility and VDAC3 mRNA abundance. Conclusion:The results of the present study implicate that zinc depletion may induce VDAC3 mRNA abundance decreasing and affect the normal function of spermatozoa.

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赵 丹,吴 炜,章 轶,王增军,王心如. Zn缺乏对人精子VDAC mRNA的影响[J].南京医科大学学报(自然科学版),2012,(4):443-448

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  • 收稿日期:2011-12-09
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