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miR-29b 影响 PROG 的分泌并促进牛黄体细胞的增殖。

MiR-29b affects the secretion of PROG and promotes the proliferation of bovine corpus luteum cells.

机构信息

Department of Laboratory Animals, College of Animal Sciences, Jilin University, Changchun, Jilin, P.R. China.

Department of Obstetrics and Gynecology, The First Hospital of Jilin University, Changchun, Jilin, P.R. China.

出版信息

PLoS One. 2018 Apr 4;13(4):e0195562. doi: 10.1371/journal.pone.0195562. eCollection 2018.

Abstract

The regulatory role of miRNAs has been explored in ovarian cells, and their effects on gonadal development, apoptosis, ovulation, steroid production and corpus luteum (CL) development have been revealed. In this study, we analyzed the expression of miR-29b at different stages of bovine CL development and predicted the target genes of miR-29b. We confirmed that miR-29b reduces the expression of the oxytocin receptor (OXTR), affects progesterone (PROG) secretion and regulates the function of the CL. RT-PCR showed that the expression of miR-29b was significantly higher in functional CL phases than in the regressed CL phase. Immunohistochemistry showed that OXTR was expressed in both large and small CL cells and was mainly located in the cell membrane and cytoplasm of these cells. We analyzed the expression levels of OXTR and found that transfection with a miR-29b mimic decreased OXTR expression, but transfection with the inhibitor had a limited effect on the expression of the OXTR protein. At the same time, the secretion of PROG was significantly increased in the miR-29b mimic-transfected group. We also analyzed the effect of miR-29b on the apoptosis of CL cells. Finally, we found that miR-29b could promote the proliferation of bovine CL cells. In conclusion, we found that miR-29b reduces the expression of OXTR and can promote PROG secretion and the proliferation of CL cells via OXTR.

摘要

miRNA 的调控作用已在卵巢细胞中得到探索,其对性腺发育、细胞凋亡、排卵、类固醇生成和黄体(CL)发育的影响也已被揭示。在这项研究中,我们分析了牛 CL 发育不同阶段 miR-29b 的表达情况,并预测了 miR-29b 的靶基因。我们证实 miR-29b 降低了催产素受体(OXTR)的表达,影响了孕激素(PROG)的分泌,并调节了 CL 的功能。RT-PCR 显示 miR-29b 在功能 CL 期的表达明显高于退化 CL 期。免疫组织化学显示 OXTR 在大、小 CL 细胞中均有表达,主要位于这些细胞的细胞膜和细胞质中。我们分析了 OXTR 的表达水平,发现转染 miR-29b 模拟物可降低 OXTR 的表达,但转染抑制剂对 OXTR 蛋白的表达影响有限。同时,miR-29b 模拟物转染组 PROG 的分泌显著增加。我们还分析了 miR-29b 对 CL 细胞凋亡的影响。最后,我们发现 miR-29b 可以促进牛 CL 细胞的增殖。总之,我们发现 miR-29b 通过 OXTR 降低 OXTR 的表达,从而促进 PROG 的分泌和 CL 细胞的增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4334/5884578/0c6d41b50cc5/pone.0195562.g001.jpg

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