Spanò M, Bartoleschi C, Cordelli E, Leter G, Tiveron C, Pacchierotti F
Division of Environmental Toxicology, ENEA Casaccia, Rome, Italy.
Cytometry. 1996 Jun 1;24(2):174-80. doi: 10.1002/(SICI)1097-0320(19960601)24:2<174::AID-CYTO10>3.0.CO;2-O.
The effects of trophosphamide on mouse reproductive cells have been investigated by flow cytometric analysis of testicular cell populations and alterations of sperm chromatin structure. Mice were treated with single intraperitoneal injections of TP, the doses ranging between 50 and 150 mg/kg, and were killed after 7, 14, 21, 28, 35, or 49 days. Dose-dependent reductions of tetraploid cells, round spermatids, and elongated spermatids were detected at 7, 21, and 28 days, respectively, reflecting cytotoxic damage to the differentiating spermatogonia compartment. The dose necessary to reduce the number of differentiating spermatogonia to half the control value was approximately 70 mg/kg. Stem cells were not affected by this treatment, and the normal spermatogenic process was restored after 7 weeks. In addition, cauda epididymal sperm were analyzed by the sperm chromatin structure assay, a flow cytometric measurement of the susceptibility of the sperm nuclear DNA to in situ acid denaturation; a statistically significant increase of sperm with altered chromatin structure was detected after a TP treatment of 150 mg/kg. Together with previous findings published in the literature, where the same doses induced heritable genetic damage, this study demonstrates a marked adverse cytotoxic effect of TP on the male reproductive integrity. All this information should be taken into consideration when TP is used in chemotherapeutic regimens.
通过对睾丸细胞群体进行流式细胞术分析以及精子染色质结构改变，研究了曲磷酰胺对小鼠生殖细胞的影响。给小鼠腹腔注射单次剂量的曲磷酰胺（TP），剂量范围为50至150mg/kg，并在7、14、21、28、35或49天后处死。分别在第7、21和28天检测到四倍体细胞、圆形精子细胞和延长型精子细胞的剂量依赖性减少，这反映了对分化中的精原细胞区室的细胞毒性损伤。将分化中的精原细胞数量减少至对照值一半所需的剂量约为70mg/kg。干细胞不受该处理的影响，并且在7周后正常的生精过程得以恢复。此外，通过精子染色质结构分析对附睾尾精子进行了分析，这是一种流式细胞术测量精子核DNA对原位酸变性敏感性的方法；在150mg/kg的TP处理后，检测到染色质结构改变的精子有统计学意义的增加。结合文献中先前发表的相同剂量诱导可遗传遗传损伤的研究结果，本研究证明了TP对雄性生殖完整性具有明显的不良细胞毒性作用。在将TP用于化疗方案时，所有这些信息都应予以考虑。
