精子过量残留胞质的特性研究

A study on the characteristics of excess residual cytoplasm in human spermatozoa

目的观察精子过量残留胞质(ERC)结构在体外的稳定性,探讨ERC的适宜检测方法及其临床意义。方法监测精子在干、湿片检测状态下精子ERC率与精液离体时间、精子活动力、活性氧类物质(ROS)以及精子成熟度指标的关系。结果 (1)精液射出体外0～120 min时间段内,无论精子处于干、湿状态,其ERC率均随精液标本离体时间延长而持续下降(P<0.001);各时间段内湿片活动精子ERC率均明显高于不活动精子(P<0.001),且ERC保有率明显高于干片(P<0.001)。(2)精液标本经上泳分离后,上泳精子湿片ERC率显著高于底层精子(P<0.001),而干片ERC水平则明显低于底层精子(P<0.001),同层精子湿片法ERC率明显高于干片法(P<0.001)。(3)湿片状态下精子ERC率与精子前向运动率、活动率无相关性(P均>0.05),但干片状态下呈显著负相关(r=-0.472、r=-0.430,P=0.009、P=0.018);(4)湿片状态下精子ERC率与ROS、核蛋白不成熟度(湿染法)、DNA碎片率(湿染法)、精子-透明质酸结合率(HBA)均无显著相关性(P>0.05);干片状态下精子ERC率与ROS(r=0.784,P<0.001)、DNA碎片(干片法)(r=0.574,P=0.001)、HBA(r=-0.336,P=0.036)显著相关,与核蛋白不成熟度(干片法)无显著相关性(P>0.05),但标本内部ERC精子较非ERC精子的核蛋白不成熟率更高(P<0.001)。结论精子ERC结构随精液离体时间延长呈进行性崩解趋势,精子在经历自然干燥后(干片法)其ERC保有率更具临床意义,干片法ERC水平与精子活动能力、ROS水平及成熟度显著相关,ERC结构是不良质量精子的标志物。

ObjectiveTo observe the stability of human spermatozoa excess residual cytoplasm （ERC） in vitro and to explore the suitable detection method of ERC and its clinical significance. MethodsThe relationship between sperm ERC rate and time in vitro of semen, sperm motility, reactive oxygen species （ROS） and sperm maturation index levels were measured by dry and wet smear detection. Results1） The ERC rate significantly decreased with the prolongation of spermatozoa in vitro for 0-120 min （P〈0.001）. The ERC rate of motile spermatozoa was significantly higher than that of immotile spermatozoa （P〈0.001）, and the ERC rate was significantly higher by wet-smear detection than by dry-smear detection （P〈0.001）. 2） After spermatozoa was selected by swim-up technique, the ERC rate of swim-up spermatozoa was significantly higher than that of underlying spermatozoa （P〈0.001） for the wet-smear detection, while it was significantly lower than that of the underlying spermatozoa （P〈0.001） for the dry-smear detection. Compared with the same layer spermatozoa, the ERC rate of wet-smear detection was higher than that of dry-smear detection （P〈0.001）. 3） There was no significant correlation between sperm ERC percentage and sperm progressive and total motility rate by wet-smear detection. But there was a significant negative correlation between sperm ERC rate and sperm progressive and total motility rate by dry-smear detection （r=-0.472, P=0.009; r=-0.430, P=0.018, respectively）. 4） There was no significant correlation between the ERC rate and ROS, immature sperm nucleoprotein （wet staining）, DNA fragmentation （wet staining） as well as the sperm-hyaluronic acid binding assay （HBA） score by wet-smear detection （P〉0.05）. However, underwent dry sperm, there were significant correlations between the ERC rate and ROS （r= 0.784, P〈0.001）, DNA fragment （dry staining） （r=0.574, P=0.001）, HBA （r=-0.336, P=0.036）, except for immature sperm nucleoprotein （dry staining）. The immature sperm nucleoprotein rate of ERC spermatozoa was higher than that of non-ERC spermatozoa after drying process （P〈0.001）. ConclusionThe ERC of spermatozoa has a tendency of progressive disintegration with the prolongation of semen in vitro. And compared with wet-smear method, the ERC retention rate of spermatozoa after natural drying （dry-smear method） has more clinical significance. The ERC level by dry-smear detection was significantly correlated with sperm motility, ROS levels and maturation. Moreover, the ERC might be a marker of poor sperm quality.