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目的体外观察血小板激活因子(PAF)对小鼠附睾精子和人成熟精子的趋化作用以及对受精的影响。②方法在微趋化室下层孔内加入27μL新鲜制备的精子悬液,上层孔内加入不同质量浓度PAF,UR-12519(PAF受体阻断剂),之间以多聚碳核孔滤网分隔,37℃孵育,用经特殊处理的玻片封闭,反转微趋化室,离心,相差显微镜下计数玻片上的精子数。用UR-12519预处理小鼠精子,通过体外受精技术观察受精率及早期胚胎发育。③结果随PAF质量浓度的增加,聚集于上层孔内的精子逐渐增多,当PAF≥1.0mg/L时,聚集的精子数量明显增多,与对照组比较,差异有极显著性(z=3.82~4.81,P<0.01);当UR-12519质量浓度增加(>1.0mg/L)时,PAF吸引精子的作用受抑。体外受精实验显示,经UR-12519预处理的精子,受精率明显低于正常,早期胚胎发育亦明显受抑。④结论PAF对小鼠附睾精子及人成熟精子有趋化作用,这种作用能被其受体阻断剂所抑制。UR-12519对小鼠精-卵体外受精及其胚胎的早期发育均有抑制作用。
Objective To observe the chemotactic effect of platelet activating factor (PAF) on mouse epididymal sperm and human mature sperm in vitro and its effect on fertilization. Methods 27μL freshly prepared sperm suspension was added into the lower chamber of micrometastasis chamber. Different concentrations of PAF and UR-12519 (PAF receptor blocker) were added to the upper wells, Separately, incubate at 37 ° C, block with specially treated slides, reverse micrometastolic chamber, centrifuge, and count the number of sperm on slides under a phase contrast microscope. Mouse sperm were pretreated with UR-12519, fertilization rate and early embryonic development were observed by in vitro fertilization technique. Results With the increase of PAF concentration, spermatozoa accumulated in the upper pores gradually increased. When the PAF≥1.0mg / L, the number of spermatozoa gathered increased significantly. Compared with the control group, the difference was significant (z = 3 .82 ~ 4.81, P <0.01). When the concentration of UR-12519 was increased (> 1.0mg / L), the effect of PAF on sperm attraction was inhibited. In vitro fertilization experiments showed that pre-treated by UR-12519 sperm, fertilization rate was significantly lower than normal, early embryo development was significantly inhibited. ④ Conclusion PAF mouse epididymal sperm and human mature sperm chemotaxis, this effect can be blocked by its receptor blocker. UR-12519 inhibits mouse sperm-egg IVF and early embryonic development.