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目的:观察碳酸酐酶Ⅱ(CA2)在人睾丸、精子中的表达定位及比较CA2在生育男性和弱精子症患者精子中的表达差异。方法:通过免疫组化观察CA2在人睾丸中的定位;通过免疫荧光观察CA2在人精子中的定位;收集健康生育男性和弱精子症患者的精液标本各16份,60%Percoll分离精液标本,排除生精细胞和白细胞,采用Western印迹方法,从蛋白水平检测CA2的表达。结果:免疫组化结果显示CA2在人睾丸中特异性定位于长形精子的尾部;免疫荧光结果显示CA2在人精子的尾部有较强表达。Western印迹结果显示弱精子症患者精子中CA2的表达显著高于生育男性(1.84±0.32vs1.41±0.26,P<0.05)。结论:CA2在睾丸生精周期的长形精子阶段才表达,定位于精子的尾部。CA2在弱精子症患者精子中表达显著增高,提示其可能与精子活动力下降有关。
Objective: To observe the expression and localization of carbonic anhydrase Ⅱ (CA2) in human testis and sperm and to compare the expression of CA2 in sperm between fertile male and asthenospermia patients. METHODS: The localization of CA2 in human testis was observed by immunohistochemistry. The localization of CA2 in human sperm was observed by immunofluorescence. Sixteen semen samples of healthy male and female patients with asthenospermia were collected. 60% Percoll semen samples were collected, Spermatogenic cells and leukocytes were excluded and Western blotting was used to detect the expression of CA2 at the protein level. Results: The results of immunohistochemistry showed that CA2 was specifically located in the tail of spermatozoa in human testis. The results of immunofluorescence showed that CA2 was highly expressed in the tail of human spermatozoa. Western blot results showed that the expression of CA2 in sperm of asthenospermia patients was significantly higher than that of males (1.84 ± 0.32 vs 1.41 ± 0.26, P <0.05). Conclusion: CA2 is only expressed in the spermatozoon stage of testicular spermatogenesis and located in the tail of sperm. The expression of CA2 in sperms of patients with asthenospermia significantly increased, suggesting that it may be related to decreased sperm motility.