非连续密度梯度法对弱精子症患者精子参数及DNA完整性的影响

Effect of discontinuous density gradient method on semen quality parameters and sperm DNA integrity in semen specimen from the asthenospermia patients

目的通过非连续密度梯度法制备弱精子症患者精液,分析制备前后精液常规分析质量参数及精子DNA完整性,评估非连续密度梯度法在改善弱精子症患者精子质量方面的有效性。方法 26例弱精子症患者来源于本院门诊,禁欲2~7d后手淫取精,使用一次性精子计数玻片(goldcyto),采用SCA精子质量分析系统对精子制备前后精子浓度、精子总活力[前向运动精子(PR)+非前向运动精子(NP)]和PR百分率进行分析;采用改良巴氏染色法分析精子形态;同时运用精子吖啶橙染色方法对制备前后的精子DNA完整性进行检测。结果 26例精液标本制备前精液体积(4.22±1.54)mL,精子浓度(51.9±44.65)×10~6/mL,精子总活力(22.5±6.34)%,PR百分率为(17.3±5.5)%,正常形态精子百分率(6.58±2.50)%,精子DNA完整率(64.96±13.33)%;制备后精子悬液体积统一调至0.5m L,精子浓度(18.91±24.61)×10~6/mL,精子总活力(31.77±21.15)%,PR百分率(26.2±20.00)%,正常形态精子百分率(7.77±3.14)%,精子DNA完整率(72.23±12.43)%。统计结果提示精子制备前后精子浓度、精子总活力、PR百分率、正常形态精子百分率和精子DNA完整率均存在显著性差异(P<0.05)。结论非连续密度梯度法制备精子能显著提高弱精子症患者精子总活力、PR百分率、正常形态精子百分率和精子DNA完整率,有效改善制备后的精子质量,为后续实施人类辅助生殖技术提供保障。

Objective To investigate the influence of discontinuous density gradient method on semen quality parameters and sperm DNA integrity in semen samples from the patients with asthenospermia, and evaluate its effectiveness in improving semen quality. Metlaoda Total of 26 male infertility patients from the outpatient department were enrolled in the study. Their semen sample were collected after a minimum of two days and a maximum of seven days of sexual abstinence. Sperm concentration, total motility （progressive motility（PR） ＋ non-progressive motility（NP）, progressive motility（PR） and sperm morphology （normal forms, %） were analyzed by SCA sperm quality analysis system before and after sperm preparation. Acridine orange staining of sperm DNA was used to detect the integrity of spermatozoa. Results The semen volume, sperm concentration, total motility（PR＋NP）, progressive motility （PR）, sperm morphology （normal forms, %） and sperm DNA integrity rate before sperm preparation were （4.22±1.54）mL, （51.9±44.65）×10^6/ mL, （22.5±6.34）%, （17.3±5.5）%, （6.58±2.50）% and （64.96±13.33）%, respectively. After sperm preparation, the volume of sperm suspension was changed into 0.5mL. Sperm concentration, total motility（PR＋NP）, progressive motility （PR）, sperm morphology （normal forms, %） and sperm DNA integrity rate were （18.91±24.61）×10^6/mL, （31.77±21.15）% （26.2±20.00）%, （7.77±3.14）% and （72.23±12.43）%, respectively. The statistical results showed that there were significant differences in sperm concentration, total motility （PR＋NP）, progressive motility （PR）, sperm morphology （normal forms, %） and sperm DNA integrity between before and after sperm preparation. Conclusion The discontinuous density gradient method can effectively improve the quality of semen from asthenospermia patients, such as total motility （PR＋NP）, progressive motility （PR）, sperm morphology （normal forms, %） and DNA integrity, which will increase the success rate of human assisted reproductive technology.