长链非编码RNA-Gm5196在高雌诱发子代脂代谢紊乱中的介导作用及机制研究

Women who have undergone ovulation stimulation therapy exhibit high serum estradiol levels throughout pregnancy. In the clinical follow-up study, we determined that their newborns were also prone to lipid metabolic disorders. However, the underlying mechanism remains unclear. With the established hyperestrogenic mouse model, we determined that the long non-coding RNA Gm5196 was associated with Osbpl5 (Oxysterol binding protein-like 5), which is an important gene in lipid metabolism; moreover, the expression of Gm5196 in the offspring’s fetal liver was decreased. Estradiol decreased Gm5196 expression, whereas it increased Osbpl5 expression in a human liver cell line. In addition, interference of Gm5196 up-regulates Osbpl5 expression and promotes the secretion of cholesterol. Here, we plan to further investigate the mediating effect and underlying mechanism of Gm5196 in high estradiol exposure-induced lipid metabolic disorders in offspring. Using a human liver cell line in culture, we will identify the key roles of Gm5196 and Osbpl5 in high estradiol interfering lipid metabolism via knockdown or overexpression experiments, define the relationship between Gm5196 and Osbpl5 using several techniques, including RIP (RNA Immunoprecipitation), and explore the estradiol transcriptional regulation of Gm5196 via the luciferase reporter system. Phenotype analysis of mouse injected with Gm5196 siRNA or overexpression plasmid will be used to further clarify the role of Gm5196 in lipid metabolism. Using the hyperestrogenic mouse model, the Gm5196 expression level in the liver of offspring was examined, and its correlation analysis with maternal serum estradiol levels was conducted. The feasibility of Gm5196 as an early biological warning indicator was also investigated through clinical investigation. Using these studies, we plan to develop ideas and provide a theoretical basis for research regarding embryonic disease induced by environmental endocrine disruptors during pregnancy.

促排卵妊娠孕妇处于高雌激素状态，我们临床随访表明子代易出现脂代谢紊乱但机制不详。借助高雌小鼠模型，我们发现脂代谢重要基因Osbpl5相关的长链非编码RNA-Gm5196在子代肝脏中表达下降；雌激素能降低肝细胞Gm5196而上调Osbpl5表达，敲减Gm5196会增加Osbpl5表达及胆固醇生成。本项目拟在此基础上深入研究Gm5196在高雌诱发子代脂代谢紊乱中的介导作用及机制。通过肝细胞系培养，敲减或过表达Gm5196和Osbpl5以明确两者在高雌干扰脂代谢中的关键作用，利用RIP等技术确定两者相互关系，借助荧光素酶报告系统分析高雌对Gm5196调控机制；通过Gm5196干扰及过表达老鼠相关分析明确其在脂代谢中作用；对高雌小鼠子代肝脏进行Gm5196表达检测，探讨其和孕鼠高雌相关性；通过临床检测探讨Gm5196作为预警指标可行性。为孕期环境内分泌干扰物致胎源性疾病研究开拓思路，提供理论基础。

本项目初步阐述了Gm5196 和OSBPL5 在肝细胞脂代谢中的重要作用和相互关系，并初步阐述了雌激素对Gm5196 转录调控的相关分子机制。这些研究结果在lncRNAs 层面上探讨孕期高雌激素暴露诱发子代脂代谢紊乱的可能致病机理，为临床诊断提供新的科学参考指标，未防治孕期子宫内不良因素暴露所导致的胚胎源性疾病提供实验依据。本项目发表了标注资助论文1篇（Fertility and Sterility, 2017）。