Guangdong Provincial Key Laboratory of Bioengineering Medicine, Department of Cell Biology, Jinan University, Guangzhou, China.
Department of Pediatrics, Peking University First Hospital, Beijing, China.
Mol Cell Endocrinol. 2019 Aug 1;493:110453. doi: 10.1016/j.mce.2019.110453. Epub 2019 May 23.
The aim of the present study was to identify microRNAs (miRNAs) that regulate the proliferation and differentiation of Leydig cells (LCs) of rat. Three small RNA libraries derived from progenitor LCs (PLCs), immature LCs (ILCs) and adult LCs (ALCs) were analyzed by microarrays. In total, 68 differentially expressed miRNAs (DEMs) were identified. Based on the trend of DEM expression from PLCs to ALCs, primary LCs were transfected with miRNA mimics or inhibitors. Five miRNAs (miR-30a-5p, miR-3585-5p, miR-212-3p, miR-369-5p and miR-434-3p) promoted PLC proliferation, and 3 miRNAs (miR-17-5p, miR-532-3p and miR-329-3p) activated caspase-3, which triggered LC apoptosis. For steroidogenesis, 18 miRNAs could elevate or inhibit androsterone release at the PLC stage. Eleven and 9 miRNAs inhibited the production of 5α-androstane-3α,17β-diol in ILCs and testosterone in ALCs, respectively. miR-17-5p, miR-29a-3p and miR-299a-5p decreased androgen production by LCs at all developmental stages. Furthermore, the miR-299a-5p-mediated decrease in androgen production by the LC lineage was primarily achieved by downregulating the expression of luteinizing hormone/choriogonadotropin receptor (LHCGR) and 3β-hydroxysteroid dehydrogenase 1 (HSD3B1). These findings provide insights into the regulatory roles of miRNAs during the postnatal development of LCs and suggest potential strategies for the treatment of steroid-related disorders.
本研究旨在鉴定调控大鼠睾丸间质细胞（LCs）增殖和分化的 microRNAs（miRNAs）。通过 microarray 分析了源自祖细胞 LC（PLC）、未成熟 LC（ILC）和成熟 LC（ALC）的三个小 RNA 文库。总共鉴定出 68 个差异表达的 miRNAs（DEMs）。基于从 PLC 到 ALC 的 DEM 表达趋势，用 miRNA 模拟物或抑制剂转染原代 LC。有 5 个 miRNAs（miR-30a-5p、miR-3585-5p、miR-212-3p、miR-369-5p 和 miR-434-3p）促进 PLC 增殖，有 3 个 miRNAs（miR-17-5p、miR-532-3p 和 miR-329-3p）激活 caspase-3，从而引发 LC 凋亡。对于类固醇生成，有 18 个 miRNAs 可以在 PLC 阶段升高或抑制雄酮的释放。有 11 个和 9 个 miRNAs 分别抑制 ILC 中 5α-雄烷-3α,17β-二醇和 ALC 中睾酮的产生。miR-17-5p、miR-29a-3p 和 miR-299a-5p 在所有发育阶段均降低 LCs 的雄激素产生。此外，LC 谱系中 miR-299a-5p 介导的雄激素产生减少主要是通过下调黄体生成素/绒毛膜促性腺激素受体（LHCGR）和 3β-羟甾脱氢酶 1（HSD3B1）的表达来实现的。这些发现为 miRNAs 在 LCs 出生后发育过程中的调控作用提供了新的认识，并为类固醇相关疾病的治疗提供了潜在策略。
