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Evaluation method for DNA damage and dysfunction of spermatogenic cells due to Bromopropanes

溴丙烷所致生精细胞 DNA 损伤和功能障碍的评价方法

基本信息

批准号：
14370124
负责人：
OKUDA Masayuki
金额：
$ 5.12万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370124/
关键词：
comet assay single cell gel electrophoresis DNA apoptosis bromopropane trichloroethylene toxicology 8-OHdG ラット精巣 DNA損傷有機溶剤 single gel electrophoresis 精原細胞温度 Syber Green

项目摘要

Comet assay (single cell gel electrophoresis) is a simple method for evaluation of DNA toxicity. DNA of rat spermatogenic cells (spermatogonia and spermatocytes) are damaged with bromopropanes in dose- and time-dependent manner. Among some kinds of bromopropanes, 1,3-bromopropane has most toxicity and 2-bromopropane and 1-boromopropane follow. Trichloroethylene, which is used as the same purposes in industry, reduces the toxicity due to 1,3-bromopropane when trichloroethylene with bromopropane is exposed at the same time to primary-cultured spermatogenic cells. But this mechanism for toxicity reduction should be further investigated. When comet assay is carried in more neutral condition than usual alkaline condition, DNA damage is not detected. DNA fragmentation due to apoptosis may not be induced with bromopropane in the condition used in this study (primary culture, Wistar rats) since rat testes DNA does not show ladder in electrophoresis. Oxidative stress markers for DNA, 8-OHdG (hydroxy-deoxy-guanosine), are elevated in DNA from testes tissue or primary-cultured spermatogenic cells. This study shows that comet assay can be used as screening method foe toxicity to testes and may differentiate DNA damage from apoptosis in some specific conditions.

彗星试验（单细胞凝胶电泳）是评价DNA毒性的一种简单方法。溴丙烷对大鼠生精细胞（精原细胞和精母细胞）DNA的损伤具有剂量和时间依赖性。在几种溴丙烷中，1，3-溴丙烷的毒性最大，其次是2-溴丙烷和1-溴丙烷。三氯乙烯，这是在工业上用于相同的目的，减少毒性由于1，3-溴丙烷时，三氯乙烯与溴丙烷同时暴露于原代培养的生精细胞。但这种毒性降低机制还有待进一步研究。当彗星试验在中性条件下进行时，没有检测到DNA损伤。在本研究中使用的条件下（原代培养，Wistar大鼠），溴丙烷可能不会诱导细胞凋亡引起的DNA片段化，因为大鼠睾丸DNA在电泳中未显示梯形。DNA的氧化应激标记物8-OHdG（羟基脱氧鸟苷）在睾丸组织或原代培养的生精细胞的DNA中升高。本研究表明，彗星试验可作为睾丸毒性的筛选方法，在某些特定条件下可鉴别DNA损伤与凋亡。