Abstract
This research was performed to estimate the potential effects of Di (2-ethylhexyl) phthalate (DEHP) on changes of ovarian miRNA expression profile during mouse primordial follicle assembly using miRNAs-seq analysis. The ovaries of newborn mice were collected and in vitro cultured with different concentration of DEHP for 72 h. Then they were prepared for miRNAs-seq analysis. The results indicated that DEHP exposure altered ovarian miRNA expression profile of newborn mice. Eighteen differentially expressed miRNAs were screened after 100 μM DEHP exposure. The target mRNAs of differentially expressed miRNAs were predicted and further analyzed through gene ontology (GO) enrichment analysis and pathway enrichment analysis. Our results showed that the differentially expressed miRNAs from DEHP exposure can regulate ovarian development by targeting mRNAs involved in MAPK, mTOR, FoxO signaling pathways. Three miRNAs of miR-32-5p, miR-19a-3p, and miR-141-3p were randomly selected from the differentially expressed miRNAs to quantify their expression level by miRNA qRT-PCR. The results of qRT-PCR and miRNA-seq were consistent. Considering one of its target gene PTEN of miR-19a-3p and the decreased level of pAKT and increased Bax/Bcl-2 under DEHP exposure, we speculated that the altered expression of miR-19a-3p by DEHP exposure affected mouse primordial follicle assembly via PI3K/AKT1/mTOR signaling pathway. Epigenetic changes are one of the most important targets of toxicant exposure. The effects of DEHP exposure on microRNA (one of the epigenetic regulators) expression profile were uncovered to enrich the research on relationship of epigenetics and toxicant exposure.
Highlights
Di (2-ethylhexyl) phthalate (DEHP) is one of the most common phthalates widely used in plastics [1] for medical apparatus and instrument, food packaging, and plastic drink bottles [2]
The concentrations of DEHP used in this study were 10 μM (3.9 μg/ml) and 100 μM (39 μg/ml), which were based on our previously dose response test and the papers published by our research group [33, 34]
Wang et al reported that both DEHP and its active metabolite mono (2-ethylhexyl) phthalate (MEHP) inhibited antral follicles growth of mouse ovaries by increased level of reactive oxygen species [35, 38]
Summary
Di (2-ethylhexyl) phthalate (DEHP) is one of the most common phthalates widely used in plastics [1] for medical apparatus and instrument, food packaging, and plastic drink bottles [2]. Diet is the largest DEHP exposure source [3], especially for fast food, perhaps because of the massive use of plastic packaging or disposable PVC gloves during fast food preparation [4]. Medical procedure is another exposure source for DEHP Impairs Ovarian miRNAs Profile patients and some of neonates [3, 5]. DEHP as an endocrine disrupting chemicals (EDC) brings reproductive toxicity to both male and female human and wild animals [3]. DEHP decreased the weight of ovary and uterus and promoted the quantity of follicles [1]
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