Zhenjiang Hospital Affiliated to Nanjing University of Chinese Medicine, Zhenjiang Hospital of Traditional Chinese Medicine, Zhenjiang, Jiangsu 212000, P.R. China.
Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China.
Int J Mol Med. 2018 Dec;42(6):3329-3343. doi: 10.3892/ijmm.2018.3874. Epub 2018 Sep 13.
The present study aimed to explore the difference in the expression profiles of ovarian microRNA sequences in rats in a light pollution environment and rats in a normal light environment. Rats in the control group were exposed to 12‑h light/dark cycles, while rats in the model group were continuously exposed to 24‑h light. The ovaries were extracted from the two groups of rats, and Illumina HiSeq 2500 high‑throughput sequencing technology was used to detect the differences in microRNA (miRNA) expression among the two groups. Fluorescence quantitative reverse transcription‑polymerase chain reaction was used to verify the differential expression of miRNA. The present study was designed to experimentally validate the interaction between miR‑421‑5p and mitogen‑activated protein kinase (MAPK) 7 by using the dual‑luciferase reporter system, and to explore the expression of proteins in the MAPK signaling pathway with a lentiviral vector‑mediated small hairpin RNA interference against microRNA‑421‑5p. The expression of 45 miRNAs was significantly different. In total, 13 miRNAs were upregulated, of which 5 miRNA sequences were known and 8 were predicted. Furthermore, 32 miRNAs were downregulated, of which 11 miRNA sequences were known and 21 were predicted. The results of the luciferase reporter assay confirmed the targeting association between miR‑421‑5p and MAPK7. The expression levels of MAPK and genes in its downstream signaling pathways, including c‑Fos, CREB and c‑Myc, were downregulated when miR‑421‑5p was overexpressed and upregulated when miR‑421‑5p was silenced. The differential expression of miRNAs may serve an important role in the development of the ovary in a light pollution environment. miR‑421‑5p may regulate ovarian growth and development by targeting the MAPK signaling pathway in light polluted rat ovaries.
本研究旨在探讨光污染环境下大鼠与正常光照环境下大鼠卵巢 microRNA 序列表达谱的差异。对照组大鼠暴露于 12 h 光照/黑暗周期，而模型组大鼠则持续暴露于 24 h 光照。从两组大鼠中提取卵巢，使用 Illumina HiSeq 2500 高通量测序技术检测两组间 microRNA（miRNA）表达的差异。采用荧光定量逆转录聚合酶链反应验证 miRNA 的差异表达。本研究旨在通过双荧光素酶报告系统实验验证 miR-421-5p 与丝裂原活化蛋白激酶（MAPK）7 之间的相互作用，并利用慢病毒载体介导的 microRNA-421-5p 小发夹 RNA 干扰技术探索 MAPK 信号通路中蛋白的表达。45 个 miRNA 的表达存在显著差异。共上调 13 个 miRNA，其中 5 个 miRNA 序列已知，8 个 miRNA 序列预测；下调 32 个 miRNA，其中 11 个 miRNA 序列已知，21 个 miRNA 序列预测。荧光素酶报告实验结果证实了 miR-421-5p 与 MAPK7 之间的靶向关联。当 miR-421-5p 过表达时，MAPK 及其下游信号通路中的基因，包括 c-Fos、CREB 和 c-Myc 的表达水平下调，当 miR-421-5p 沉默时表达水平上调。miRNA 的差异表达可能在光污染环境下卵巢的发育中发挥重要作用。miR-421-5p 可能通过靶向光污染大鼠卵巢中的 MAPK 信号通路来调节卵巢的生长和发育。
