Li Yan, Zhang Hao, Liang Ying, Wu De-sheng
Department of Environmental Health, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 Jul;37(4):562-5.
To study the effects of Flutamide (Flu) on the full gene expression of early testis development of embryonic period in male SD rats.
Flu was injected subcutaneously (sc) with NS at doses of 6.25 mg/kg x d from 12th to 17th gestation days (GD), and in control group with tales doses NS. Fifty pregnant SD rats were sacrificed on GD 20. The mRNA were extracted respectively from fetus testis with treated and untreated Flu, then transcribed reversely to cDNA and labeled with cy5 and cy3 fluorescence. Subsequently, cDNA probes were hybridized to the Mouse40S cDNA microarray and fluorescence signals were analyzed. Meanwhile, we took out of the right testes of male offspring to make the detection of immunohistochemistry (IHC). Furthermore, we determined the level of testosterone by enzyme-linked immunosorbentassay (ELISA).
Compared with the control group, a total of 31 identified genes expressed differently, including 14 down-regulated and 17 upregulated genes. However, the function of 5 down-regulated and 10 upregulated difference expression genes are unknown in Genbank. This finding has provided important reference basis for the molecular mechanism of reproductive development of Flu in the future. The activity of two important enzymes was cut down by IHC. The level of testosterone was decreased by ELISA.
The results indicated that Flu administrated to F0 pregnant rats can damage apparently to the reproductive development of embryonic period male SD rats in many ways, and can cause the abnormality of sexual differentiation and sexual development in rats.
研究氟他胺（Flu）对雄性SD大鼠胚胎期睾丸早期发育全基因表达的影响。
于妊娠第12至17天，将氟他胺以6.25mg/kg×d的剂量与生理盐水皮下注射，对照组注射等量生理盐水。妊娠第20天处死50只妊娠SD大鼠。分别从经氟他胺处理和未处理的胎鼠睾丸中提取mRNA，然后逆转录为cDNA并分别用cy5和cy3荧光标记。随后，将cDNA探针与Mouse40S cDNA微阵列杂交并分析荧光信号。同时，取出雄性子代的右侧睾丸进行免疫组织化学（IHC）检测。此外，通过酶联免疫吸附测定（ELISA）测定睾酮水平。
与对照组相比，共鉴定出31个差异表达基因，其中14个下调，17个上调。然而，在Genbank中5个下调和10个上调的差异表达基因功能未知。这一发现为今后氟他胺生殖发育的分子机制提供了重要参考依据。免疫组织化学检测显示两种重要酶的活性降低。酶联免疫吸附测定显示睾酮水平降低。
结果表明，对F0代妊娠大鼠给予氟他胺可在多方面明显损害胚胎期雄性SD大鼠的生殖发育，并可导致大鼠性分化和性发育异常。
