不同浓度帕瑞昔布钠对大鼠离体精子运动功能、体外获能和顶体反应的影响

Effects of different concentrations of parecoxib sodium on rat sperm motility, capacitation and acrosome reaction in vitro

目的 评价不同浓度帕瑞昔布钠对大鼠离体精子运动功能、体外获能和顶体反应的影响.方法 通过扩散法收集SD大鼠附睾精子,采用随机数字表法分为4组（n=18）：对照组（C组）、100、500和1 000μmol/L帕瑞昔布钠组（P1-3组）.在37℃孵箱中培养5h后,采用计算机辅助精子分析系统评价精子运动功能,记录精子运动活力[（a＋b）％]、平均速度（VAP）、直线速度（VSL）、曲线速度（VCL）和侧摆幅度（ALH）,采用金霉素荧光染色技术评价精子体外获能情况,采用考马斯亮蓝染色法评价精子顶体反应情况.结果 C组、P1-3组精子VAP、VSL、VCL和精子获能率依次降低（P＜0.05）.与C组比较,P2.3组（a＋b）％和P2组ALH降低（P＜0.05）.各组精子顶体反应率比较差异无统计学意义（P＞0.05）.结论 帕瑞昔布钠呈浓度依赖性抑制大鼠离体精子运动功能和体外获能,对顶体反应无明显影响.

Objective To evaluate the effects of different concentrations of parecoxib sodium on the rat sperm motility,capacitation and acrosome reaction in vitro.Methods The sperm samples from Sprague-Dawley rat epididymis were collected by Klinefelter diffusion method and randomly divided into 4 groups （n =18 each） using a random number table：control group （group C）,and parecoxib sodium 100,500,1 000 μmol/L groups （P1-3 groups）.Parecoxib sodium with the final concentrations of 100,500 and 1 000 μmol/L was added to the culture medium.The samples were then incubated for 5 h in an airtight container filled with 5 % CO2 at 37 ℃.Then sperm motility was examined in vitro at 37 ℃ and analyzed by the computer-assisted sperm analysis,including the sperm motility （（a ＋ b）%）,average path velocity （VAP）,straight line velocity （VSL）,curvilinear velocity （VCL） and amplitude of lateral head displacement （ALH）.The capacitation effect was assessed by using the chlortetracycline staining and phase-contract microscopy.The acrosome reaction was evaluated by coomassie brilliant blue staining.Results The VAP,VSL,VCL and capacitation ability of the sperm were gradually decreased in C and P1-3 groups （P 〈 0.05）.Compared with group C,（a ＋ b）% in P2,3 groups and ALH in P2 group were significantly decreased （P 〈 0.05）.There was no significant difference in the acrosome reaction between groups （P 〉 0.05）.Conclusion Parecoxib sodium has significant inhibitory effects on the rat sperm motility and capacitation in a dose-dependent manner,while has no effect on the acrosome reaction in vitro.