Toxic effects of octylphenol on cultured rat spermatogenic cells and Sertoli cells.
Toxic effects of octylphenol on cultured rat spermatogenic cells and Sertoli cells.
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辛基苯酚对培养的大鼠生精细胞和支持细胞的毒性作用。
DOI：
10.1006/taap.1999.8664
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Millette,CF
中科院分区：
文献类型：
--
作者：
Raychoudhury,SS;Blake,CA;Millette,CF
烷基酚类化合物，包括雌激素类化合物4-叔辛基苯酚(OP)，属于环境污染物。由于OP对成年雄性大鼠生精功能有损害作用，且OP在体外对水生动物和哺乳动物脾细胞有毒性作用，因此我们研究了OP对培养的雄性大鼠生精细胞和支持细胞是否有直接毒性作用。用活/死Eukolight活性/细胞毒性试剂盒检测细胞活性。用含10−8M OP的成年大鼠混合生精细胞或19~21日龄大鼠支持细胞与10−12 M OP混合培养，但不加0.08%乙醇、10−6 M 17β雌二醇(E_2)或地塞米松(地塞米松)处理24 h后，存活率显著降低。所有处理都没有显著改变细胞总数。生精细胞的流式细胞仪分析显示，10−4或10−6M OP可产生异常的相对倍性。10−6M OP处理4小时后，10−6 M E_2或地塞米松处理4小时后，支持细胞染色质明显凝聚。当支持细胞在无钙条件下培养时，10−6M OP作用24 h后细胞存活率仍有下降趋势。支持细胞与10−6M OP孵育2 4h后，显微镜下可见3‘-OHDNA末端标记的胞核变小，而不与赋形剂或10−6M E2或地塞米松孵育2 4h。结果表明，OP对培养的大鼠生精细胞和支持细胞有直接毒性作用，而不是E2或DEX，提示OP对支持细胞的这种毒性作用是通过钙非依赖性的细胞凋亡来实现的。
Alkylphenols, including the estrogenic 4-tert-octylphenol (OP), are environmental pollutants. Because administration of OP to adult male rats impairs spermatogenesis and OP has been shown to be toxic to aquatic animals and to mammalian splenocytes in vitro, we studied whether OP exerts direct toxic effects on cultured spermatogenic cells and Sertoli cells isolated from male rats. Cell viability was assessed with a Live/Dead Eukolight viability/cytotoxicity kit. Culture of mixed spermatogenic cells from adult rats with 10−8M OP or Sertoli cells from 19- to 21-day-old rats with 10−12M OP, but not with 0.08% EtOH (vehicle) or 10−6M 17β-estradiol (E2) or dexamethasone (DEX), significantly decreased the percentage of viable cells after 24 h of treatment. None of the treatments significantly altered total cell number. Flow cytometric analyses of spermatogenic cells revealed that exposure to 10−4or 10−6M OP yielded abnormal relative ploidy classes. Four hours of treatment with 10−6M OP, but not with 10−6M E2or DEX, caused significant chromatin condensation in Sertoli cells as observed with acridine orange staining. The decreased percentage of viable cells after 24 h of exposure to 10−6M OP remained when Sertoli cells were cultured in Ca2+-free medium. Sertoli cells contained nuclei of reduced size and labeled 3′-OH DNA ends as detected by microscopic analyses when the cells had been incubated for 24 h with 10−6M OP but not with vehicle or 10−6M E2or DEX. The results demonstrate that OP, but not E2or DEX, is directly toxic to cultured rat spermatogenic cells and Sertoli cells and suggest that this toxic effect in Sertoli cells is exerted through Ca2+-independent apoptosis.