Wang Bin, Xia Lin, Zhu Dan, Zeng Hongtao, Wei Bin, Lu Likui, Li Weisheng, Shi Yajun, Liu Jingliu, Zhang Yunfang, Sun Miao
Institute for Fetology, The First Affiliated Hospital of Soochow University, Suzhou City, China.
Medical Center of Hematology, The Xinqiao Hospital of Army Medical University, Chongqing, China.
Front Mol Biosci. 2022 Apr 11;9:857875. doi: 10.3389/fmolb.2022.857875. eCollection 2022.
Paternal lifestyle, stress and environmental exposures play a crucial role in the health of offspring and are associated with non-genetic inheritance of acquired traits, however the underlying mechanisms are unclear. In this study, we aimed to find out how the sperm tsRNA involved in paternal high-fat diet induced abnormal gluconeogenesis of F1 offspring, and explore the underlying molecular mechanism of its regulation. We generated a paternal high fat diet (42% kcal fat) model to investigate the mechanism by which paternal diet affects offspring metabolism. Four-week-old C57BL/6J male mice were randomly assigned into two groups to receive either a control diet (CD; 10% kcal fat) or a high-fat (HFD; 42% kcal fat) diet for 10 weeks, and mice from each group were then mated with 8-week-old females with control diet in a 1:2 ratio to generate F1. F0 sperms were isolated and small RNAs was sequenced by high-throughput sequencing. Metabolic phenotypes were examined with both F0 and F1. A significant increase in body weight was observed with HFD-F0 mice at 8 weeks of age as compared to CD mice at the same age. F0 mice showed impaired glucose tolerance (GTT), resistance to insulin tolerance (ITT) and increased pyruvate tolerance (PTT) at 14 weeks. HFD-F1 male mice showed no significant difference in body weight. An increase in PTT was found at 13 weeks of age and no significant changes in GTT and ITT. PEPCK and G6Pase that related to gluconeogenesis increased significantly in the liver of HFD-F1 male mice. Sperm sequencing results showed that 5'tsRNA-Gly-GCC derived from tRNA-Gly-GCC-2 specifically was remarkably upregulated in sperm of HFD F0 mice. Q-PCR further showed that this tsRNA was also increased in the liver of HFD-F1 comparison with CD-F1 mice. In addition, we found that 5'tsRNA-Gly-GCC can regulate Sirt6-FoxO1 pathway and be involved in the gluconeogenesis pathway in liver. 5'tsRNA-Gly-GCC that increased in HFD mice mature sperms can promote gluconeogenesis in liver by regulating Sirt6-FoxO1 pathway, which might represent a potential paternal epigenetic factor mediating the intergenerational inheritance of diet-induced metabolic alteration.
父系生活方式、压力和环境暴露对后代健康起着至关重要的作用，并与获得性性状的非遗传继承有关，然而其潜在机制尚不清楚。在本研究中，我们旨在找出精子tsRNA如何参与父系高脂饮食诱导的F1代后代异常糖异生，并探索其调控的潜在分子机制。我们建立了父系高脂饮食（42%千卡脂肪）模型，以研究父系饮食影响后代代谢的机制。将四周龄的C57BL/6J雄性小鼠随机分为两组，分别给予对照饮食（CD；10%千卡脂肪）或高脂（HFD；42%千卡脂肪）饮食10周，然后将每组小鼠以1:2的比例与食用对照饮食的八周龄雌性小鼠交配，以产生F1代。分离F0代精子，并通过高通量测序对小RNA进行测序。对F0代和F1代的代谢表型进行检测。与同龄的CD小鼠相比，八周龄的HFD-F0小鼠体重显著增加。14周龄时，F0小鼠表现出葡萄糖耐量受损（GTT）、胰岛素耐量抵抗（ITT）和丙酮酸耐量增加（PTT）。HFD-F1雄性小鼠体重无显著差异。13周龄时发现PTT增加，GTT和ITT无显著变化。与糖异生相关的磷酸烯醇式丙酮酸羧激酶（PEPCK）和葡萄糖-6-磷酸酶（G6Pase）在HFD-F1雄性小鼠肝脏中显著增加。精子测序结果显示，来源于tRNA-Gly-GCC-2的5'tsRNA-Gly-GCC在HFD F0小鼠精子中特异性显著上调。定量聚合酶链反应（Q-PCR）进一步显示，与CD-F1小鼠相比，该tsRNA在HFD-F1小鼠肝脏中也增加。此外，我们发现5'tsRNA-Gly-GCC可以调节沉默调节蛋白6（Sirt6）-叉头框蛋白O1（FoxO1）通路，并参与肝脏中的糖异生途径。在HFD小鼠成熟精子中增加的5'tsRNA-Gly-GCC可以通过调节Sirt6-FoxO1通路促进肝脏中的糖异生，这可能代表一种潜在的父系表观遗传因素，介导饮食诱导的代谢改变的代际遗传。
