RNA Sequencing of Mouse Liver Post Caloric Resrtiction and Anti-Anging Interventions

Caloric restriction (CR) extends lifespan in mammals but the stringent 20% reduction in food intake is challenging to sustain; therefore, validating pharmacological mimics is of high interest. It has been previously reported that a SERPINE1 mutation in an Amish cohort links to a 10-year lifespan increase and improved metabolism. SERPINE1 encodes for Plasminogen Activator Inhibitor-1 (PAI-1) which is a clinical predictor of vascular stiffness, hypertension, and diastolic dysfunction. Moreover, PAI-1 amplifies accumulation of senescent cells as a component of the senescence associated secretory phenotype (SASP). Here, we compared hepatic transcriptomic profiles in mice after 8 weeks of CR, rapamycin, metformin, or the PAI-1 inhibitor TM5614 versus an ad libitum control. CR induced extensive changes (1087 genes downregulated, 1145 upregulated), while TM5614 showed the closest mimicry (211 down, 218 up), sharing ~50% of CR's differentially expressed genes, including downregulated Il11ra1 and upregulated Per2, tied to longevity and metabolism. Rapamycin and metformin showed minimal overlap with CR. Gene Ontology analysis revealed CR and TM5614 downregulated metabolic pathways, and both countered gene-length-dependent transcription decline, an emerging measure of an aging transcriptomic landscape. TM5614 presents as a CR mimic, offering an alternative to CR to extend healthspan. Overall design: Liver transcriptomes were analyzed from 30 C57BL/6J mice following 8 weeks of intervention beginning at 21 weeks of age. Mice were assigned to one of five groups: ad libitum control (n=6), 30% caloric restriction (CR) (n=3), TM5614 (20 mg/kg/day) (n=3), rapamycin (42 mg/kg/day) (n=3), or metformin (37.5 mg/kg/day) (n=3), with compounds incorporated into chow. At the end of the study, whole-liver RNA was extracted for sequencing (reported here in the GEO Submission).

热量限制（Caloric restriction, CR）可延长哺乳动物的寿命，但严格的20%食物摄入量限制方案难以长期坚持，因此验证药理学模拟剂的研究具有重要意义。
此前有研究报道，阿米什人群队列中的SERPINE1基因突变与寿命延长10年及代谢改善相关。
SERPINE1基因编码纤溶酶原激活物抑制剂-1（Plasminogen Activator Inhibitor-1, PAI-1），该蛋白是血管僵硬、高血压及舒张功能障碍的临床预测因子。
此外，PAI-1作为衰老相关分泌表型（Senescence Associated Secretory Phenotype, SASP）的组成部分，可促进衰老细胞的积累。
本研究中，我们对比了小鼠在接受8周热量限制、雷帕霉素、二甲双胍或PAI-1抑制剂TM5614干预后的肝脏转录组谱，并以自由进食组作为对照。
热量限制诱导了广泛的基因表达变化（1087个基因下调，1145个基因上调），而TM5614的模拟效果最为接近（211个基因下调，218个上调），二者共享约50%的差异表达基因，其中包括与寿命及代谢相关的下调基因Il11ra1和上调基因Per2。
雷帕霉素与二甲双胍与热量限制的基因表达重叠度极低。
基因本体（Gene Ontology, GO）分析显示，热量限制与TM5614均可下调代谢通路，且二者均能抵抗基因长度依赖性转录水平下降——这是衰老转录组特征的新兴衡量指标。
TM5614可作为热量限制的模拟剂，为延长健康寿命提供了替代方案。
研究整体设计：本研究对30只21周龄开始接受8周干预的C57BL/6J小鼠的肝脏转录组进行分析。小鼠被分为5组：自由进食对照组（n=6）、30%热量限制组（CR，n=3）、TM5614干预组（20 mg/kg/天，n=3）、雷帕霉素干预组（42 mg/kg/天，n=3）及二甲双胍干预组（37.5 mg/kg/天，n=3），受试化合物均掺入饲料中。实验结束后，提取全肝RNA进行测序（本数据已提交至GEO数据库）。