Cui Zhaolei, Chen Liangyuan, Liu Yaohua, Zeng Zhangxin, Lan Fenghua
Department of Clinical Genetics and Experimental Medicine, Fuzong Clinical College, Fujian Medical University, Fuzhou 350025, Fujian, China.
Department of Clinical Genetics and Experimental Medicine, Fuzong Clinical College, Fujian Medical University, Fuzhou 350025, Fujian, China.
Acta Histochem. 2015 Apr;117(3):235-42. doi: 10.1016/j.acthis.2015.02.009. Epub 2015 Mar 18.
The enzyme activity of lactate dehydrogenase C4 (LDH-C4, due to tetrameric nature of C-subunit) has been proposed as an important parameter in evaluating sperm motility and semen quality. A novel histochemical staining method for detecting LDH-C4 activity in human spermatozoa is described in this report. The staining working solution comprises sodium 2-hydroxybutyrate (an affinity substrate of LDH-C4), nitrotetrazolium blue chloride (NBT), nicotinamide adenine dinucleotide (NAD) and naphthol blue. The positive products were purple black lumps concentrated in the neck segment of the spermatozoa and weakly in the middle piece. A normal reference range for the integral enzyme activity was constructed from 120 healthy males based upon the scoring criteria. The study further compared the staining method with the routine spectrophotometry technique in terms of the results of 96 cases with infertile status. Moreover, we found the down-regulated LDH-C4 expression was significantly correlated with the lowered enzyme activity (r=0.865, P=0.000). Our data suggest that the histochemical staining method hallmarks a relatively high accuracy and may be a better alternative for measuring LDH-C4 activity in human spermatozoa.
乳酸脱氢酶C4（LDH - C4，由于C亚基的四聚体性质）的酶活性已被提议作为评估精子活力和精液质量的一个重要参数。本报告描述了一种用于检测人类精子中LDH - C4活性的新型组织化学染色方法。染色工作液包括2 - 羟基丁酸钠（LDH - C4的亲和底物）、氯化硝基四氮唑蓝（NBT）、烟酰胺腺嘌呤二核苷酸（NAD）和萘酚蓝。阳性产物为紫黑色块状物，集中在精子的颈部，中段较弱。根据评分标准，基于120名健康男性构建了整体酶活性的正常参考范围。该研究进一步在96例不育患者的结果方面，将染色方法与常规分光光度法技术进行了比较。此外，我们发现LDH - C4表达下调与酶活性降低显著相关（r = 0.865，P = 0.000）。我们的数据表明，组织化学染色方法具有较高的准确性，可能是测量人类精子中LDH - C4活性的更好选择。
