Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Livestock, Ministry of Agriculture (MOA), Urumqi, 830026, Xinjiang, China.
Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, 830026, Xinjiang, China.
J Ovarian Res. 2024 Jan 2;17(1):3. doi: 10.1186/s13048-023-01328-6.
The intricate interplay of gene expression within ovarian granulosa cells (GCs) is not fully understood. This study aimed to investigate the miRNA regulatory mechanisms of ferroptosis during the process of follicle development in lamb GCs.
Employing transcriptome sequencing, we compared differentially expressed mRNAs (DE-mRNAs) and miRNAs (DE-miRNAs) in GCs from lambs treated with follicle-stimulating hormone (FL) to untreated controls (CL). We further screened differentially expressed ferroptosis-related genes and identified potential miRNA regulatory factors. The expression patterns of HMOX1 and miRNAs in GCs were validated using qRT‒PCR and Western blotting. Additionally, we investigated the regulatory effect of oar-miR-134-3p on HMOX1 and its function in ferroptosis through cell transfection and erastin treatment.
We identified a total of 4,184 DE-mRNAs and 304 DE-miRNAs. The DE-mRNAs were mainly enriched in ferroptosis, insulin resistance, and the cell cycle. Specifically, we focused on the differential expression of ferroptosis-related genes. Notably, the ferroptosis-related genes HMOX1 and SLC3A2, modulated by DE-miRNAs, were markedly suppressed in FLs. Experimental validation revealed that HMOX1 was significantly downregulated in FL and large follicles, while oar-miR-134-3p was significantly upregulated compared to that in the CLs. HMOX1 expression was regulated by the targeting effect of oar-miR-134-3p. Functional assays further revealed that modulation of oar-miR-134-3p influenced HMOX1 expression and altered cellular responses to ferroptosis induction by erastin.
This study suggested that oar-miR-134-3p and HMOX1 may be one of the pathways regulating ferroptosis in GCs. This finding provides new clues to understanding the development and regulatory process of follicles.
卵巢颗粒细胞（GCs）中基因表达的复杂相互作用尚未完全阐明。本研究旨在研究羔羊 GC 中卵泡发育过程中 ferroptosis 的 miRNA 调控机制。
采用转录组测序，比较卵泡刺激素（FL）处理和未处理对照（CL）的羔羊 GC 中差异表达的 mRNA（DE-mRNAs）和 miRNA（DE-miRNAs）。进一步筛选差异表达的 ferroptosis 相关基因，并鉴定潜在的 miRNA 调控因子。采用 qRT-PCR 和 Western blot 验证 HMOX1 和 GC 中 miRNAs 的表达模式。此外，通过细胞转染和 erastin 处理研究 oar-miR-134-3p 对 HMOX1 的调控作用及其在 ferroptosis 中的功能。
共鉴定出 4184 个 DE-mRNAs 和 304 个 DE-miRNAs。DE-mRNAs 主要富集在 ferroptosis、胰岛素抵抗和细胞周期中。具体来说，我们关注 ferroptosis 相关基因的差异表达。值得注意的是，受 DE-miRNAs 调节的 ferroptosis 相关基因 HMOX1 和 SLC3A2 在 FL 中明显受到抑制。实验验证表明，FL 和大卵泡中 HMOX1 表达明显下调，而 oar-miR-134-3p 表达明显上调。HMOX1 表达受 oar-miR-134-3p 的靶向调节。功能测定进一步表明，oar-miR-134-3p 的调节影响 HMOX1 的表达，并改变细胞对 erastin 诱导 ferroptosis 的反应。
本研究表明 oar-miR-134-3p 和 HMOX1 可能是调节 GC 中 ferroptosis 的途径之一。这一发现为理解卵泡的发育和调控过程提供了新的线索。
