Genistein up-regulates miR-20a to disrupt spermatogenesis via targeting Limk1.
Genistein up-regulates miR-20a to disrupt spermatogenesis via targeting Limk1.
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金雀异黄素通过靶向 Limk1 上调 miR-20a 以破坏精子发生
DOI：
10.18632/oncotarget.17637
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发表时间：
2017-08-29
期刊：
影响因子：
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通讯作者：
Wang X
中科院分区：
文献类型：
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作者：
Gu H;Wu W;Yuan B;Tang Q;Guo D;Chen Y;Xia Y;Hu L;Chen D;Sha J;Wang X
染料木黄酮（GEN）是一种对雄性生殖有影响的黄酮类化合物。然而，其潜在机制仍不清楚。miRNAs是一类在精子发生过程中起重要作用的非编码小分子RNA。我们检测了不育受试者的GEN水平和miR-17-92簇表达，发现miR-17-92可能参与GEN诱导的精子发生异常。为了澄清，我们用不同剂量的GEN（0、0.5、5、50和250 mg/kg/天）喂养成年ICR小鼠35天，以研究潜在的机制。结果发现，GEN 5 mg/kg/d灌胃组小鼠精子平均路径速度、直线运动速度和欧洲运动速度均显著降低，相应剂量组小鼠睾丸组织中miR-17和miR-20 a表达水平升高。我们还发现miR-20 a是唯一在人和小鼠中差异表达的miRNA。应用生物信息学方法预测Limk 1为miR-20 a的靶基因，miR-20 a与精子发生有关。Limk 1在相应组中显著降低。双荧光素酶报告实验也证明miR-20 a可以直接靶向Limk 1。提示Limk 1可能是GEN诱导的精子发生异常中miR-20 a的靶基因。
Genistein (GEN) is one of the isoflavones that has effect on male reproduction. However, the underlying mechanism remains unknown. miRNAs are a type of small non-coding RNAs that play important roles in spermatogenesis. We measured the GEN levels and miR-17-92 cluster expression in infertile subjects and found that miR-17-92 might be involved in GEN induced abnormal spermatogenesis. To clarify, we fed adult ICR mice with different doses of GEN (0, 0.5, 5, 50 and 250 mg/kg/day) for 35 days to study the underlying mechanism. We found that sperm average path velocity, straight-line velocity and eurvilinear velocity of the mice orally with GEN at 5mg/kg/day were significantly decreased, the expression levels of miR-17 and miR-20a in mice testis were higher in corresponding group. We also found miR-20a was the only miRNA that differentially expressed both in human and mice. By applying bioinformatics methods, Limk1 was predicted to be the target gene of miR-20a that is involved in spermatogenesis. Limk1 were significantly decreased in the corresponding group. Dual-luciferase report assay also proved that miR-20a could directly target Limk1. These results implied that Limk1 might be the target gene of miR-20a that is involved in GEN induced abnormal spermatogenesis.