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.