Expression of FKBP52 in the ovaries of PCOS rats
Expression of FKBP52 in the ovaries of PCOS rats
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FKBP52在PCOS大鼠卵巢中的表达
DOI：
10.3892/ijmm.2018.3998
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发表时间：
2019-02-01
影响因子：
5.4
通讯作者：
Tan, Yong
中科院分区：
文献类型：
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作者：
Song, Shiyan;Tan, Yong
本研究旨在检测多囊卵巢综合征（PCOS）大鼠卵巢组织中FK-506结合蛋白52（FKBP52）的表达及其通过丝裂原激活蛋白激酶（MAPK）/细胞外信号调节激酶（ERK）途径介导雄激素受体（AR）的作用。采用脱氢表雄酮注射法建立PCOS模型大鼠。酶联免疫吸附测定（ELISA）测量血清性激素。苏木精和伊红（H＆E）染色用于检查卵巢组织的组织学变化。通过免疫组织化学（IHC）染色、逆转录定量聚合酶链反应（RT-qPCR）分析和蛋白质印迹（WB）检测FKBP52的表达水平。另外，采用RT-qPCR分析检测AR的mRNA表达量，采用WB检测AR、ERK1/2和磷酸化(p-)ERK1/2的蛋白表达水平。在颗粒细胞 (GC) 实验中，提取并培养原代 GC。 FKBP4是FKBP52编码基因，因此构建腺病毒载体Ad-Oe-FKBP4-EGFP和Ad-siRNA-FKBP4-EGFP，利用RT-qPCR和WB方法检查上述因素之间的关联。动物实验中，阴道涂片、H&E染色及ELISA结果表明PCOS模型建立成功。 IHC染色显示PCOS模型组GC中FKBP52的表达高于其余各组（P<0.01）。 PCOS模型大鼠FKBP52和AR的mRNA和表达水平较其他大鼠显着升高（P<0.05）。 p-ERK1/2的表达水平也较高(P<0.05)。 GC实验中，FKBP4基因过表达后，FKBP52和AR的mRNA和表达水平增加（P<0.05）。 p-ERK1/2的表达水平也增加（P<0.05）。 FKBP4基因沉默后，FKBP52和AR的mRNA和表达水平降低（P<0.05）。 ERK1/2表达水平也降低(P<0.05)。然而，p-ERK1/2的表达水平升高（P<0.05）。总之，共伴侣FKBP52的上调可能通过MAPK/ERK途径介导AR的激活。
The present study aimed to examine the expression of FK-506 binding protein 52 (FKBP52) in the ovary tissues of rats with polycystic ovarian syndrome (PCOS) and its action on mediating androgen receptor (AR) through the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway. PCOS model rats were established by dehydroepiandrosterone injection. Enzyme-linked immunosorbent assay (ELISA) measured serum sex hormones. Hematoxylin and eosin (H&E) staining was used to examine histological changes of the ovarian tissues. The expression levels of FKBP52 were detected by immunohistochemical (IHC) staining, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis and western blotting (WB). In addition, RT-qPCR analysis was used to detect the mRNA expression of AR, and WB was used to detect the protein expression levels of AR, ERK1/2 and phosphorylated (p-) ERK1/2. In granulosa cell (GC) experiments, primary GCs were extracted and cultured. FKBP4 is the FKBP52-encoding gene, therefore, adenovirus vectors Ad-Oe-FKBP4-EGFP and Ad-siRNA-FKBP4-EGFP were constructed to examine the association among the above factors using the RT-qPCR and WB methods. In the animal experiment, the vaginal smear, H&E staining and ELISA results showed that the PCOS model was successfully established. The IHC staining revealed that the expression of FKBP52 in the GCs of the PCOS model group was higher than the remaining groups (P<0.01). The mRNA and expression levels of FKBP52 and AR in the PCOS model rats were significantly increased, when compared with levels in the other rats (P<0.05). The expression level of p-ERK1/2 was also higher (P<0.05). In the GC experiment, following overexpression of the FKBP4 gene, the mRNA and expression levels of FKBP52 and AR were increased (P<0.05). The expression level of p-ERK1/2 was also increased (P<0.05). Following FKBP4 gene silencing, the mRNA and expression levels of FKBP52 and AR were decreased (P<0.05). The expression level of ERK1/2 was also decreased (P<0.05). However, the expression level of p-ERK1/2 was increased (P<0.05). In conclusion, the upregulation of co-chaperone FKBP52 may mediate the activation of AR through the MAPK/ERK pathway.