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目的:体外观察小鼠富含半胱氨酸分泌蛋白-1(mCRISP 1)抗体对受精过程中各环节的影响,探讨mCRISP 1特异性抗体的抗生育作用机制。方法:获取重组mCRISP 1蛋白疫苗免疫后8周小鼠的免疫血清,梯度稀释(1∶10,1∶50,1∶100,1∶200)后与生育功能正常的小鼠精子悬液共孵育,对照组采用未免疫血清。观察mCRISP 1抗体对精子存活率、活力、凝集反应、顶体反应及对精卵穿透试验的影响。结果:各组间精子的存活率、活力、顶体反应发生率均无显著差异(P>0.05),mCRISP 1抗血清与正常雄性小鼠精子共孵育后,精子无凝集现象;当抗血清浓度≥1∶100时,精卵融合率显著下降(P<0.05),但各组间每卵黏附精子数无显著差异(P>0.05)。结论:mCRISP 1抗体主要通过抑制小鼠精卵融合过程降低小鼠生育能力。
OBJECTIVE: To observe the effect of mCRISP-1 antibody on the various stages of fertilization in vitro and to investigate the anti-fertility mechanism of mCRISP-1-specific antibody. Methods: The immune serum of mice immunized with recombinant mCRISP 1 protein vaccine was obtained after 8 weeks of immunization. The mice were incubated with sperm suspension of normal fertility mice after gradient dilution (1:10, 1:50, 1:100, 1:200) , The control group with non-immune serum. The effects of mCRISP 1 antibody on sperm viability, viability, agglutination, acrosome reaction and sperm penetration test were observed. Results: There was no significant difference in sperm motility, acrosome reaction and acrosome reaction among groups (P> 0.05). There was no agglutination of sperm after incubation with mCRISP1 antiserum in sperm of normal male mice. When antiserum concentration ≥1: 100, the sperm-egg fusion rate was significantly decreased (P <0.05), but there was no significant difference in the number of adherent sperm between the groups (P> 0.05). Conclusion: The mCRISP 1 antibody can reduce the fertility of mice primarily by inhibiting the sperm-egg fusion process in mice.