ABSTRACT

1. Granulosa cells (GCs) are involved in folliculogenesis, follicular development, and atresia. Previous studies have shown that microRNA-181a-5p (miR-181a-5p) and sirtuin 1 (SIRT1) are involved in GC proliferation and apoptosis, and SIRT1 has been predicted as one target of miR-181a-5p. However, there are little studies with poultry.

2. Quantitative real-time PCR (qRT-PCR) was used to detect the expression level of miR-181a-5p in granulosa layers during geese ovarian follicular development. A methyl thiazolyl tetrazolium (MTT) assay was performed to assess the viability of geese granulosa cells treated with miR-181a-5p mimic or inhibitor. The binding sites between the SIRT1 3′-UTR region and miR-181a-5p were evaluated using a luciferase reporter assay system. SIRT1 mRNA levels were detected using qRT-PCR after transfection with miR-181a-5p mimic and inhibitor.

3. The miR-181a-5p suppressed geese GC viability and regulated the mRNA expression of viability-related genes in geese GCs. SIRT1 was a target gene of miR-181a-5p and miR-181a-5p suppressed its mRNA expression.

4. The miR-181a-5p may target and inhibit SIRT1 expression, thus suppressing GC viability by regulating viability-related key genes.

摘要

1.颗粒细胞（GC）参与卵泡形成，卵泡发育和闭锁。先前的研究表明microRNA-181a-5p（miR-181a-5p）和sirtuin 1（SIRT1）参与GC增殖和凋亡，并且SIRT1被预测为miR-181a-5p的靶标之一。但是，关于家禽的研究很少。

2.定量实时PCR（qRT-PCR）用于检测鹅卵泡发育过程中颗粒层中miR-181a-5p的表达水平。进行了甲基噻唑基四唑鎓（MTT）分析，以评估用miR-181a-5p模拟物或抑制剂处理的鹅颗粒细胞的活力。使用萤光素酶报告基因检测系统评估SIRT1 3'-UTR区与miR-181a-5p之间的结合位点。用miR-181a-5p模拟物和抑制剂转染后，使用qRT-PCR检测SIRT1 mRNA水平。

3. miR-181a-5p抑制了鹅GC的活力，并调节了鹅GC中与活力相关的基因的mRNA表达。SIRT1是miR-181a-5p的靶基因，miR-181a-5p抑制了其mRNA表达。

4. miR-181a-5p可能靶向并抑制SIRT1表达，从而通过调节生存力相关关键基因来抑制GC生存力。