孕期DEHP暴露后子代雌鼠下丘脑lncRNA的差异性表达研究

Differential expression of IncRNA in the hypothalamus of offspring female rats after DEHP uterus exposure

目的 探究孕期DEHP暴露对大鼠子代雌鼠下丘脑长链非编码RNA(lncRNA)表达的影响,分析并预测DEHP影响子代下丘脑-垂体-性腺轴与能量代谢的机制。方法 大鼠孕0~18 d连续灌胃染毒,子代雌鼠出生后立刻取下丘脑组织并提取总RNA,使用illumina Hiseq测序平台进行测序,筛选出差异表达的mRNA和lncRNA进行GO与KEGG富集分析。利用实时荧光定量PCR检测技术验证染毒组与对照组的下丘脑细胞中差异表达的目的lncRNA。结果 通过RNA-seq测序分析,筛查到108条差异表达的lncRNA和342条差异表达的mRNA。KEGG分析显示差异表达的lncRNA主要富集在雌激素信号通路、Ras信号通路信号转导和PI3K-Akt信号通路等通路。结论 孕期DEHP暴露后子代雌鼠下丘脑组织存在lncRNA及mRNA的差异表达,且差异表达的lncRNA和mRNA功能分析结果显示产前DEHP暴露可能对子代雌鼠下丘脑的性腺激素和能量代谢过程有影响。

Objective To investigate the effect of DEHP uterus exposure on the expression of lncRNA in the hypothalamus of offspring female rats,and to analyze and predict the mechanism by which DEHP affects the hypothalamic pituitary gonadal axis and energy metabolism in offspring.Methods Rats were exposed to DEHP continuously by gavage from 0 to 18 days since pregnant.The hypothalamic tissue of the offspring female rats was immediately taken after birth and total RNA was extracted.The Illumina Hiseq sequencing platform was used for sequencing,and differentially expressed mRNA and lncRNA were screened for GO and KEGG enrichment analysis.Results The results showed that 108 significantly differentially expressed lncRNA and 342 significantly differentially expressed mRNA were identified through RNA-seq sequencing analysis.KEGG analysis showed that differentially expressed lncRNA was mainly enriched in estrogen signaling pathway,Ras signaling pathway signal transduction,and PI3K-Akt signaling pathway.Conclusion There are differentially expressed lncRNA and mRNA in hypothalamic tissue of female offspring after DEHP exposure during pregnancy,and the functional analysis results of differentially expressed lncRNA and mRNA indicate that prenatal DEHP exposure may affect the gonadal hormone and energy metabolism process in hypothalamus of female offspring.