不育症精子乳酸脱氢酶同功酶LDHx活性测定及其定位研究

DETERMINATION OF ACTIVITY AND STUDY OF LOCALIZATION ABOUT LACTATE DEHYDROGENASE ISOENZYME LDHx IN INFERTILE SPERMATOZOA

采用聚丙烯酰胺凝胶电泳、酶联染色、光密度扫描、分光光度法以及电镜酶细胞化学等方法 ,对 12例生育男性(生育组 )和 14例不育男性 (不育组 )精子 L DHx进行了研究。结果显示 L DHx电泳区带位于 L DH3和 L DH4之间 ,生育组精子 L DHx绝对活性和相对活性均高于不育组 (P<0 .0 5 )。相关分析表明精子密度与 L DHx绝对活性和相对活性均具有相关性 ,在生育组呈正相关 (r=0 .8和 0 .75 ,P<0 .0 5 ) ,在不育组呈负相关 (r=- 0 .76和 - 0 .78,P<0 .0 5 )。 L DHx酶细胞化学定位分析显示 L DHx酶反应颗粒主要分布于精子型线粒体 (STM)和胞质内 ,少量分布于顶体及质膜表面。生育组精子各部位酶反应颗粒多于不育组 ,且不育组精子多有畸形并伴有超微结构改变。上述研究分析提示 ,精子 L DHx活性测定与定位分析可作为检查不育症精子质量的可靠指标 。

The methods of polyacrylamide gel electrophoresis (PAGE), enzyme linked staining, desitometric scanning, spectrophotometry and electron microscopic enzyme cytochemistry were used to stduy LDHx activity of sperm obtained from 12 fertile men and 14 infertile men. The results showed that the 'X band' appeared in the major area of LDH between LDH 3 and LDH 4 on lactate dehydrogenase isoenzyme electrophoretic pattern. The absolute and relative activity of spermatozoon LDHx in fertile group were higher than that of infertile group ( P <0 05). In fertile group, the absolute and relative activity of spermatozoon LDHx were related positively to sperm concentration ( r =0 8 and 0 75,respectively, P <0 05). But in infertile group, this relationship showed inverse correlation ( r =-0 76 and -0 78,respectively, P <0 05). Electron microscopic cytochemistry proved that the enzymic particles of LDHx were mainly localizated in the matrix of sperm type mitochondria, acrosome, cytoplasm and plasma membrane and some escaped out of membrane. The number of enzymic particles of LDHx in fertile spermatozoon was apparently larger than those in infertile spermatozoon. Moreover, spermatozoon from infertile men mostly showed deformity and a change of spermatic ultrastructure. These results suggested that the examining of LDHx activity and the study of the distribution of LDHx may be used as a reliable marker for testing the quality of spermatozoa and diagnosing the male sterility.