密度梯度离心联合上游法对精子DNA损伤的影响

Effects of Density Gradient Centrifugation Combined withSwim-Up on Sperm DNA Damage

目的探讨不同精液优化处理方法对精子DNA碎片指数(DNA fragmentation index,DFI)和高DNA着色性(high DNA stainability,HDS)的影响。方法收集30例来本生殖中心就诊男性患者的新鲜精液样本,采用上游法、密度梯度离心法和密度梯度离心联合上游法3种方法优化精液,通过精子染色质结构分析检测新鲜精液及3种优化方法处理后的精子DFI和HDS水平。结果新鲜精液的精子DFI水平为(15.35±1.33)%,HDS水平为(2.69±0.27)%,前向运动精子百分比为(34.87±2.27)%,处理后3组的精子DFI和HDS水平均显著降低(P<0.05),前向运动精子百分比均显著提高(P<0.05)。密度梯度离心联合上游组的精子DFI水平最低,前向运动精子百分比最高,且与上游组和密度梯度离心组间差异具有统计学意义(P<0.05)。上游组和密度梯度离心组的精子DFI水平和前向运动精子百分比差异无统计学意义(P>0.05)。密度梯度离心联合上游组和上游组的HDS水平显著低于密度梯度离心组(P<0.05),两组间差异无统计学意义(P>0.05)。结论密度梯度离心联合上游处理可极大程度降低精液中精子DFI和HDS水平,获得活力更好的精子,是一种更为理想的精液优选方法。

Objective To investigate effects of different semen optimizing methods on sperm DNA fragmentation index(DFI)and high DNA stainability(HDS).Methods Fresh semen samples of 30 male patients were collected at the Reproductive Medicine Center of Zhuozhou City Hospital.Three methods were used to optimize semen,including swim-up,density gradient centrifugation and density gradient centrifugation combined with swim-up.We detected DFI and HDS levels in fresh semen and sperm treated with three optimized methods by analyzing sperm chromatin structure.Results DFI level of fresh semen was(15.35±1.33)%,while HDS level was(2.69±0.27)%,and the percentage of PR was(34.87±2.27)%.DFI and HDS levels of sperm in all three groups after treatment were significantly decreased(P<0.05),and PR sperm was significantly increased(P<0.05).DFI level of sperm in the density gradient centrifugation combined with swim-up group was the lowest,and PR sperm was the highest,which was significantly different from the swim-up group and density gradient centrifugation group(P<0.05).There were no significant differences in DFI level and percentage of PR sperm between the swim-up group and density gradient centrifugation group(P>0.05).HDS level of the swim-up group and the density gradient centrifugation combined with swim-up group was significantly lower than that of the density gradient centrifugation group(P<0.05),and there was no significant difference between the two groups(P>0.05).Conclusion Density gradient centrifugation combined with swim-up can greatly reduce the level of DFI and HDS in sperm and obtain sperm with a better motility,which is a more applicable method for semen optimization.