Molecular mechanisms of atrazine toxicity on H19-7 hippocampal neurons revealed by integrated miRNA and mRNA “omics”

Abstract

Atrazine (ATR) is a widely applied herbicide in Asia and South America with slow natural degradation and documented deleterious effects on human and animal health, including hippocampal toxicity. However, relatively little is known about the molecular mechanisms responsible for ATR-induced hippocampal damage. Screening for differentially expressed mRNAs and microRNAs (miRNAs), and construction of potential miRNA-mRNA regulatory networks can reveal such mechanisms, so we analyzed the mRNA and miRNA expression profiles of rat hippocampus-derived H19–7 cells in response to ATR (500 μM) and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes enrichment (KEGG) analyses. Integration of miRNA sequencing (miRNA-seq) and mRNA sequencing (mRNA-seq) results identified 114 differentially expressed miRNAs (DEMIs, 40 upregulated and 74 downregulated), and 510 differentially expressed mRNAs (DEMs, 177 upregulated and 333 downregulated) targeted by these DEMIs. The top 10 hub mRNAs (Fos, Prkcb, Ncf1, Vcam1, Atf3, Pak3, Pak1, Cacna1s, Junb, and Ccl2) and 19 related miRNAs (rno-miR-194–5p, rno-miR-24–3p, rno-miR-3074, rno-miR-1949, rno-miR-218a-1–3p, rno-miR-1843a-5p, rno-miR-1843b-5p, rno-miR-296–3p, rno-miR-320–3p, rno-miR-219a-1–3p, rno-miR-122–5p, rno-miR-1839–5p, rno-miR-1843a-3p, rno-miR-215, rno-miR-3583–3p, rno-miR-194–3p, rno-miR-128–1–5p, rno-miR-1956–5p, and rno-miR-466b-2–3p) were validated by quantitative real-time PCR. GO analysis indicated that these DEMs were enriched in genes associated with synaptic plasticity and antioxidant capacity, while KEGG analysis suggested that enriched DEMs were involved in calcium signaling, axon guidance, MAPK signaling, and glial carcinogenesis. The miRNA-mRNA regulatory network identified here may provide potential biomarkers and novel strategies for the treatment of hippocampal neurotoxicity induced by ATR.