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目的:探讨弱精子症患者精子中富含半胱氨酸的分泌蛋白2(CRISP2)mRNA及蛋白的表达水平,探讨其与精子活力的关系及相关的分子机制。方法:收集成年男性弱精子症患者精液78例,活力正常精液70例作为对照组,50% Percoll离心分离纯化后,提取精子总RNA及总蛋白,采用RT-PCR,SYBR Green实时定量PCR和Western印迹技术检测CRISP2 mRNA及蛋白相对表达量。结果:CRISP2基因在弱精子症患者精子中的表达量明显低于其在正常对照组精子中的表达量,下降达4.3倍;Western印迹发现CRISP2蛋白在弱精子症组较正常对照组下降达1.71倍,两组之间的表达量有明显的统计学差异(P<0.05)。结论:CRISP2基因及蛋白在弱精子症患者精子中的表达下调可能与精子活力下降有密切联系,提示CRISP2基因及蛋白可能成为弱精子症发病机制研究的一个分子靶标。
OBJECTIVE: To investigate the expression of cysteine-rich secreted protein 2 (CRISP2) mRNA and protein in sperms of asthenospermia patients and to explore its relationship with sperm motility and related molecular mechanisms. Methods: Seventy-eight cases of adult male patients with asthenospermia were collected, and 70 cases of normal sperm were used as control group. Total RNA and total protein of sperm were extracted by centrifugation and purification with 50% Percoll. RT-PCR, SYBR Green real-time PCR and Western The relative expression of CRISP2 mRNA and protein was detected by Western blotting. Results: The expression of CRISP2 in spermatozoa of asthenospermia patients was significantly lower than that in normal sperms, which was 4.3-fold lower than that in control sperms. Western blotting showed that CRISP2 protein in asthenospermia group decreased by 1.71 Fold, there was a significant difference between the two groups (P <0.05). CONCLUSIONS: The down-regulation of CRISP2 gene and protein in sperms of asthenospermia patients may be closely related to the decline of sperm motility, suggesting that CRISP2 gene and protein may be one of the molecular targets for studying the pathogenesis of asthenospermia.