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目的采用FQ-PCR扩增不育小白鼠睾丸转铁蛋白基因,探讨腺嘌呤致昆明小白鼠雄性不育的可能机制。方法用含0.5%的腺嘌呤饲料饲喂小白鼠5～10d,分别在实验第5、10d取睾丸组织,常规方法切片,HE染色后观察病理变化。根据转铁蛋白基因序列设计、合成引物,荧光定量PCR检测实验组与对照组转铁蛋白基因表达水平。结果第5、10d实验组小白鼠睾丸组织Tf基因水平差异有统计学意义(t=5.178,P<0.01);对照组5周末Tf基因表达量是实验组的2.51倍(t=2.783,P<0.05),第10周的表达量是实验组的6.59倍(t=11.864,P<0.01)。结论腺嘌呤致雄性不育小鼠睾丸转铁蛋白基因表达水平降低。转铁蛋白基因可作为不育症诊断及治疗的重要靶标。
Objective To investigate the possible mechanism of male sterility caused by adenine in Kunming mice by FQ-PCR amplification of testis transferrin gene in mice. Methods The mice were fed with 0.5% adenine diet for 5 ~ 10 days. Testicular tissues were collected on the 5th and 10th day of the experiment. The sections were sliced by conventional method. The pathological changes were observed by HE staining. According to the sequence of transferrin gene, primers were designed and the expression levels of transferrin gene in experimental and control groups were detected by fluorescence quantitative PCR. Results The level of Tf gene in testis tissue of the 5th and 10th day in experimental group was significantly different (t = 5.178, P <0.01). The expression of Tf gene in the control group was 2.51 times that of the experimental group (t = 2.783, P < 0.05). The expression level in the 10th week was 6.59 times that in the experimental group (t = 11.864, P <0.01). Conclusion Adenine-induced male infertility testis transferrin gene expression decreased. Transferrin gene can be used as an important target for diagnosis and treatment of infertility.