Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China.
State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China.
Reproduction. 2021 May;161(5):523-537. doi: 10.1530/REP-20-0568.
Long ncRNAs regulate a complex array of fundamental biological processes, while its molecular regulatory mechanism in Leydig cells (LCs) remains unclear. In the present study, we established the lncRNA LOC102176306/miR-1197-3p/peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) regulatory network by bioinformatic prediction, and investigated its roles in goat LCs. We found that lncRNA LOC102176306 could efficiently bind to miR-1197-3p and regulate PPARGC1A expression in goat LCs. Downregulation of lncRNA LOC102176306 significantly supressed testosterone (T) synthesis and ATP production, decreased the activities of antioxidant enzymes and mitochondrial complex I and complex III, caused the loss of mitochondrial membrane potential, and inhibited the proliferation of goat LCs by decreasing PPARGC1A expression, while these effects could be restored by miR-1197-3p inhibitor treatment. In addition, miR-1197-3p mimics treatment significantly alleviated the positive effects of lncRNA LOC102176306 overexpression on T and ATP production, antioxidant capacity and proliferation of goat LCs. Taken together, lncRNA LOC102176306 functioned as a sponge for miR-1197-3p to maintain PPARGC1A expression, thereby affecting the steroidogenesis, cell proliferation and oxidative stress of goat LCs. These findings extend our understanding of the molecular mechanisms of T synthesis, cell proliferation and oxidative stress of LCs.
长链非编码 RNA 调节着一系列复杂的基本生物学过程，但其在间质细胞（LCs）中的分子调节机制尚不清楚。在本研究中，我们通过生物信息学预测建立了 lncRNA LOC102176306/miR-1197-3p/过氧化物酶体增殖物激活受体γ共激活因子 1α（PPARGC1A）调控网络，并研究了其在山羊 LCs 中的作用。我们发现 lncRNA LOC102176306 可以有效地与 miR-1197-3p 结合，并调节山羊 LCs 中的 PPARGC1A 表达。下调 lncRNA LOC102176306 显著抑制了睾酮（T）的合成和 ATP 的产生，降低了抗氧化酶和线粒体复合物 I 和复合物 III 的活性，导致线粒体膜电位丧失，并通过降低 PPARGC1A 的表达抑制了山羊 LCs 的增殖，而 miR-1197-3p 抑制剂处理可恢复这些作用。此外，miR-1197-3p 模拟物处理显著缓解了 lncRNA LOC102176306 过表达对 T 和 ATP 产生、抗氧化能力和山羊 LCs 增殖的正向作用。综上所述，lncRNA LOC102176306 作为 miR-1197-3p 的海绵，维持了 PPARGC1A 的表达，从而影响了山羊 LCs 的类固醇生成、细胞增殖和氧化应激。这些发现扩展了我们对 T 合成、LCs 细胞增殖和氧化应激分子机制的理解。
